CN114854641A - Low-temperature-resistant composite denitrifying bacteria and application thereof in sewage treatment - Google Patents
Low-temperature-resistant composite denitrifying bacteria and application thereof in sewage treatment Download PDFInfo
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- 241000894006 Bacteria Species 0.000 title claims abstract description 59
- 239000002131 composite material Substances 0.000 title claims abstract description 41
- 239000010865 sewage Substances 0.000 title claims abstract description 24
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 56
- 241000589774 Pseudomonas sp. Species 0.000 claims abstract description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 28
- 241001478271 Rahnella aquatilis Species 0.000 claims abstract description 15
- 238000004321 preservation Methods 0.000 claims abstract description 5
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims abstract description 4
- 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 25
- JVMRPSJZNHXORP-UHFFFAOYSA-N ON=O.ON=O.ON=O.N Chemical compound ON=O.ON=O.ON=O.N JVMRPSJZNHXORP-UHFFFAOYSA-N 0.000 claims description 21
- 239000001963 growth medium Substances 0.000 claims description 17
- 241000589516 Pseudomonas Species 0.000 claims description 14
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 3
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims description 2
- 230000004913 activation Effects 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 239000000243 solution Substances 0.000 claims description 2
- 230000001580 bacterial effect Effects 0.000 abstract description 40
- 230000015556 catabolic process Effects 0.000 abstract description 28
- 238000006731 degradation reaction Methods 0.000 abstract description 28
- 230000007613 environmental effect Effects 0.000 abstract description 8
- 244000005700 microbiome Species 0.000 abstract description 8
- 238000000034 method Methods 0.000 abstract description 4
- 239000000126 substance Substances 0.000 abstract description 3
- 239000002351 wastewater Substances 0.000 abstract 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract 1
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 244000144972 livestock Species 0.000 abstract 1
- 239000001301 oxygen Substances 0.000 abstract 1
- 229910052760 oxygen Inorganic materials 0.000 abstract 1
- 244000144977 poultry Species 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 45
- 230000000694 effects Effects 0.000 description 25
- 150000001875 compounds Chemical class 0.000 description 16
- 238000011081 inoculation Methods 0.000 description 12
- 238000012258 culturing Methods 0.000 description 10
- 239000002609 medium Substances 0.000 description 8
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical group N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 7
- 238000005259 measurement Methods 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000010802 sludge Substances 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 238000013329 compounding Methods 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000001546 nitrifying effect Effects 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000010485 coping Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
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- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
Classifications
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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
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
-
- 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
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/166—Nitrites
-
- 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
-
- 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
-
- 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
- C12R2001/38—Pseudomonas
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The invention belongs to the technical field of biological denitrification of sewage and environmental microorganisms, and particularly relates to a low-temperature-resistant composite denitrifying bacterium and an application thereof, wherein the composite denitrifying bacterium is a mixture of at least two strains of Rahnella aquatilis strain WS33, Pseudomonas sp (Pseudomonas sp.) strain DT04 and Pseudomonas sp (Pseudomonas sp.) strain DT 06. Can be applied to the degradation removal of high-concentration inorganic nitrogen pollution and Chemical Oxygen Demand (COD) in sewage under the low-temperature condition. The bacterial strains WS33, DT04 and DT06 are all preserved in China general microbiological culture Collection center (CGMCC) with the preservation numbers of CGMCC No.22997, 22998 and 22999. The composite bacteria can be used for various water treatment processes of medium-high concentration domestic sewage, livestock and poultry breeding wastewater or industrial organic wastewater and the like in cold regions in China, and has the advantages of wide applicable temperature range, environmental friendliness and the like.
Description
Technical Field
The invention belongs to the technical field of environmental microbiology and biological sewage treatment, and particularly relates to low-temperature-resistant composite denitrifying bacteria and application thereof in sewage treatment.
Background
In the world, the increase of the concentration of inorganic nitrogen such as ammoniacal nitrogen and nitrate nitrogen in water such as surface water can cause a series of problems of eutrophication and the like and the deterioration of ecological environment. How to effectively remove inorganic nitrogen pollution and organic matter pollution in the polluted water body has important practical significance.
Microorganisms represented by nitrifying bacteria, denitrifying bacteria and the like can utilize inorganic nitrogen pollutants and COD in the sewage and finally convert most of the pollutants into harmless nitrogen and CO 2 And removing the harmless substances from the water body (biological denitrification technology for short). Compared with physical and chemical treatment technologies, the biological denitrification technology has the advantages of economy, feasibility, environmental friendliness and the like, and is a mainstream process method for sewage denitrification and COD degradation widely adopted by countries in the world at present.
For COD degradation and denitrification microorganisms, their physiological activities are often affected by temperature. Under the condition of low temperature, the denitrification effect and the COD removal effect of the microorganisms are inhibited and reduced. Most denitrifying microorganisms such as nitrifying bacteria and denitrifying bacteria are mesophilic bacteria, that is, the optimal temperature range for the microorganisms to exert denitrifying activity is 25-35 ℃. However, when the temperature is lowered in autumn and winter, the activity and the quantity of the denitrifying microorganisms are reduced, so that the denitrifying efficiency and the COD degradation effect of a sewage system are obviously reduced, and the effluent quality is unstable or does not reach the standard.
The discovery of the denitrifying bacterial strain which can endure the low temperature condition to improve the denitrifying and COD degrading effects of the sewage in the low temperature season has important significance. A large amount of active exploration is conducted by a plurality of research and development institutions and enterprise units at home and abroad around the screening of high-efficiency strain resources and the optimization of a biological treatment process. At present, most of the existing researches are single bacteria, the adaptability of the single bacteria to complicated water quality components and variable environmental conditions of sewage is weak, and particularly, the single bacteria is sensitive to impact caused by water quality fluctuation and the like to further cause poor treatment effect.
On one hand, most of the existing biological denitrification and COD degradation strain resources are mesophilic bacteria, the adaptability to low-temperature conditions below 25 ℃ is poor, and the denitrification activity and the COD degradation are low. The existing low-temperature-resistant microbial resources are extremely lack, and the treatment requirements of denitrification and COD degradation of the polluted water body in cold seasons cannot be better met; on the other hand, the single low temperature resistant denitrifying bacteria have weak ability to adapt to complex water quality compositions and environmental condition fluctuation, and it is necessary to improve the environmental adaptability and functional stability of microorganisms, thereby ensuring continuous denitrification and COD degradation performance.
Disclosure of Invention
The invention aims to provide a low-temperature-resistant composite denitrifying bacterium and application thereof in sewage treatment, aiming at the current situations of poor biological denitrification effect in low-temperature seasons of sewage, lack of low-temperature denitrification and COD degradation strain resources and weak capability of a single strain in coping with complex environmental conditions.
In order to achieve the purpose, the invention adopts the technical scheme that:
a low temperature resistant composite denitrifier is a mixture of at least two strains of Rahnella aquatilis (Rahnella aquatilis) strain WS33, Pseudomonas sp (Pseudomonas sp.) strain DT04, and Pseudomonas sp (Pseudomonas sp.) strain DT 06.
The composite denitrobacteria are Rahnella aquatilis strain WS33, Pseudomonas (Pseudomonas sp.) strain DT04 and Pseudomonas (Pseudomonas sp.) strain DT 06.
The composite denitrifying bacteria are Rahnella aquatilis strain WS33, Pseudomonas (Pseudomonas sp.) strain DT04 and Pseudomonas (Pseudomonas sp.) strain DT06, and the number ratio of the strains is 2-5:2-8: 1-3.
The Pseudomonas (Pseudomonas sp.) strain DT04, the Rahnella aquatilis (Rahnella aquatilis) strain WS33 and the Pseudomonas (Pseudomonas sp.) strain DT06 are all preserved in the China general microbiological culture Collection center (CGMCC) at 8 and 2 months in 2021, the preservation numbers of the strains are CGMCC No.22998, 22997 and 22999 respectively, and the preservation address is West Lu No. 1 institute 3 of North Chen of the Yangtze district in Beijing.
An application of the low temperature resistant composite denitrifying bacteria and an application of the low temperature resistant composite denitrifying bacteria in sewage treatment.
The low-temperature resistant composite denitrifying bacteria are applied to sewage treatment at the temperature of 6-30 ℃.
The low-temperature resistant composite denitrifying bacteria are respectively activated by using liquid culture media, and the activation culture conditions are as follows: shaking at 80-180rpm at 20-35 deg.C for 24-48 hr with initial pH of the culture medium of 7.0-8.0, mixing the activated strains, and adding into the polluted water to be treated.
The liquid culture medium comprises the following components: NaCl 0.5-12.0; peptone 0.4-12.0; 0.2-6.0 of yeast extract and 1000mL of distilled water, and the pH is adjusted to 7.2-8.0.
Mixing the activated bacteria solutions, and mixing the mixed solution with a Rahnella aquatilis strain WS33, a Pseudomonas sp (Pseudomonas sp) strain DT04 and a Pseudomonas sp (Pseudomonas sp) strain DT06 according to the number ratio of the strains of 2-5:2-8: 1-3.
When the above-mentioned materials are mixed, the cell number in every bacterial liquor is not less than 10 8 One/ml.
Inoculating the composite bacterial liquid into the inorganic nitrogen polluted water body to be treated according to the inoculation amount of 0.5-10%.
The form of the inorganic nitrogen in the polluted water body is ammonia nitrogen, nitrate nitrogen and nitrite nitrogen; wherein the concentration range of ammoniacal nitrogen is 10-300mg/L, the concentration range of nitrate nitrogen is 5-800mg/L, the concentration range of nitrite state is 0.2-100mg/L, and the concentration range of COD is 50-4000 mg/L.
The salinity (by NaCl concentration) of the sewage is 0.1-30g/L, and the pH value of the sewage is 6.0-9.0.
The compound microbial inoculum is adopted to realize the effective removal of inorganic nitrogen and COD within 3 to 10 days.
The invention has the beneficial effects that:
(1) the compound bacteria are beneficial to improving the tolerance to temperature, salinity and pollutant load;
(2) the composite bacteria can well adapt to low-temperature conditions, and can effectively remove COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in water at 6 ℃;
(3) the strain keeps higher denitrification activity in a wide temperature range of 6-40 ℃, and can be used for biological denitrification treatment of sewage in a wide geographical area with large annual temperature difference change in the north of China;
(4) the strain has good growth and reproduction capability under the condition of medium and low temperature, and the strain propagation and the seed liquid preparation are relatively easy, thereby being beneficial to industrial production and subsequent application.
Detailed Description
The present invention will be further described with reference to the following examples, but the present invention is not limited to the following examples.
The 3 strains of bacteria of the composite bacteria are selected from activated sludge of a sewage treatment plant in Heilongjiang, belong to Laenhei and Pseudomonas by primary identification, and have obvious degradation and removal effects on inorganic nitrogen and COD in a water body under a low temperature condition.
Materials required in the following examples:
1) activated sludge: collected from an activated sludge tank of a sewage treatment plant in Heilongjiang province.
2) Culture medium:
enrichment culture medium: 10.0 parts of NaCl; 10.0 parts of peptone; and 5.0 of yeast extract.
Denitrogenation medium (g/L): CH (CH) 3 COONa 8.2；NH 4 Cl 0.50、KNO 3 0.58、NaNO 2 0.24、KH 2 PO 4 0.08；CaCl 2 ·2H 2 O 0.1；MgSO 4 ·7H 2 O 1.0；FeSO 4 ·7H 2 O0.006; 1mL of trace element liquid; 1L of distilled water; the pH value is 7.0-7.5; solid medium was supplemented with 2% agar.
Microelement liquid (g/L): 0.35 parts of EDTA; ZnSO 4 ·7H 2 O 0.2；CuSO 4 ·5H 2 O 0.1； MnSO 4 ·7H 2 O 0.1；Co(NO 3 ) 2 ·7H 2 O 0.09；H 3 BO 3 0.1；Na 2 MoO 4 0.1。
Sterilizing the above culture medium in a sterilizing pot at 103.4kPa and 121 deg.C for 20min, and cooling.
3) An experimental instrument:
a constant temperature incubator, a constant temperature shaking incubator, an ultra-clean workbench, a vertical pressure steam sterilizer, a gel imaging system, a micro ultraviolet spectrophotometer, a PCR instrument, an electrophoresis apparatus, a vortex oscillator and the like.
Example 1
(first) screening of Strain
Taking 20g of activated sludge, putting the activated sludge into a 250mL conical flask which is pre-filled with 100mL of enrichment medium, and then carrying out shake culture on a constant-temperature shaking table at 10 ℃ and 180rpm for 48 h;
after 48 hours, taking the upper layer bacteria liquid after standing, and coating the upper layer bacteria liquid in a denitrification culture medium flat plate after 10 times of gradient dilution;
after bacterial colonies on a screening culture medium grow, picking single bacterial colonies by using an aseptic inoculating loop, inoculating the single bacterial colonies on a denitrification solid culture medium by adopting a plate streaking separation method, culturing the single bacterial colonies in a 10 ℃ constant-temperature incubator for 48 hours, picking the single bacterial colonies after the bacterial colonies grow out, repeating the streaking separation process until the bacterial colonies are single in shape, and respectively taking bacterial strains with the maximum removal rates of ammonium nitrogen, nitrate nitrogen and COD through denitrification effect determination, wherein the bacterial strains are respectively provided with the numbers of WS33, DT04 and DT 06.
(II) identifying strain and preserving strain
The genomic DNA of WS33, DT04 and DT06 were extracted separately and sent to Beijing Liu-He Hua Dai Gene science and technology Co., Ltd for 16S rRNA gene sequencing. Through sequence alignment, the closest genetic relationship between the strain WS33 and Rahnella aquatica and the closest genetic relationship between DT04 and DT06 and pseudomonas are preliminarily determined. The 16S rRNA gene sequences of the strains are respectively as follows:
WS33:
TGGCTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGCAGCGGAAAGTAGC TTGCTACTTTGCCGGCGAGCGGCGGACGGGTGAGTAATGTCTGGGAAACTGCCTGATGGAGGGG GATAACTACTGGAAACGGTAGCTAATACCGCATGACCTCGAAAGAGCAAAGTGGGGGATCTTCG GACCTCACGCCATCGGATGTGCCCAGATGGGATTAGCTAGTAGGTGAGGTAATGGCTCACCTAG GCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGAC TCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGC GTGTGTGAAGAAGGCCTTAGGGTTGTAAAGCACTTTCAGCGAGGAGGAAGGCATCATACTTAAT ACGTGTGGTGATTGACGTTACTCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGT AATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTTGTTA AGTCAGATGTGAAATCCCCGCGCTTAACGTGGGAACTGCATTTGAAACTGGCAAGCTAGAGTCT TGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGT GGCGAAGGCGGCCCCCTGGACAAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGA TTAGATACCCTGGTAGTCCACGCTGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTG GCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAA TGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAAC CTTACCTACTCTTGACATCCACGGAATTCGCCAGAGATGGCTTAGTGCCTTCGGGAACCGTGAG ACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGC GCAACCCTTATCCTTTGTTGCCAGCACGTGATGGTGGGAACTCAAAGGAGACTGCCGGTGATAA ACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGAGTAGGGCTACACACGTGC TACAATGGCATATACAAAGAGAAGCGAACTCGCGAGAGCAAGCGGACCTCATAAAGTATGTCGT AGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTAGATCAGA ATGCTACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGA
16S rRNA gene sequence of strain DT 04:
CCCGAAGGTTAGACTAGCTACTTCTGGTGCAACCCACTCCCATGGTGTGACGGGCGGTGTGTAC AAGGCCCGGGAACGTATTCACCGCGACATTCTGATTCGCGATTACTAGCGATTCCGACTTCACG CAGTCGAGTTGCAGACTGCGATCCGGACTACGATCGGTTTTCTGGGATTAGCTCCACCTCGCGG CTTGGCAACCCTCTGTACCGACCATTGTAGCACGTGTGTAGCCCAGGCCGTAAGGGCCATGATG ACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCCTTAGAGTGCCCACCATT ACGTGCTGGTAACTAAGGACAAGGGTTGCGCTCGTTACGGGACTTAACCCAACATCTCACGACA CGAGCTGACGACAGCCATGCAGCACCTGTCTCAATGTTCCCGAAGGCACCAATCTATCTCTAGA AAGTTCATTGGATGTCAAGGCCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTC CACCGCTTGTGCGGGCCCCCGTCAATTCATTTGAGTTTTAACCTTGCGGCCGTACTCCCCAGGC GGTCAACTTAATGCGTTAGCTGCGCCACTAAGAGCTCAAGGCTCCCAACGGCTAGTTGACATCG TTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCACCTCAGTGT CAGTATCAGTCCAGGTGGTCGCCTTCGCCACTGGTGTTCCTTCCTATATCTACGCATTTCACCG CTACACAGGAAATTCCACCACCCTCTACCATACTCTAGCTCGTCAGTTTTGAATGCAGTTCCCA GGTTGAGCCCGGGGATTTCACATCCAACTTAACGAACCACCTACGCGCGCTTTACGCCCAGTAA TTCCGATTAACGCTTGCACCCTCTGTATTACCGCGGCTGCTGGCACAGAGTTAGCCGGTGCTTA TTCTGTCGGTAACGTCAAAATTGCAGAGTATTAATCTACAACCCTTCCTCCCAACTTAAAGTGC TTTACAATCCGAAGACCTTCTTCACACACGCGGCATGGCTGGATCAGGCTTTCGCCCATTGTCC AATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTCTCAGTTCCAGTGTGACTGATC ATCCTCTCAGACCAGTTACGGATCGTCGCCTTGGTGAGCCATTACCCCACCAACTAGCTAATCC GACCTAGGCTCATCTGATAGCGCAAGGCCCGAAGGTCCCCTGCTTTCTCCCGTAGGACGTATGC GGTATTAGCGTCCGTTTCCGAACGTTATCCCCCACTACCAGGCAGATTCCTAGGCATTACTCAC CCGTCCGCCGCTCTCAAGAGAAGCAAGCTTCTCTCTACCGCTCGACTTGCATGTGTTAGTCTG
16S rRNA gene sequence of strain DT 06:
CTCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGTAGAGAGAAGCTTGCTT CTCTTGAGAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGGTAGTGGGGGATAACGTT CGGAAACGGACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCG CTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATC CGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGG AGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAA GAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCAGTTACCTAATACGTGATTG TTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAG GGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGGATG TGAAATCCCCGGGCTCAACCTGGGAACTGCATTCAAAACTGACTGACTAGAGTATGGTAGAGGG TGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGC GACCACCTGGACTAATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACC CTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGAAGCCTTGAGCTTTTAGTGGCGCA GCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGAC GGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGG CCTTGACATCCAATGAACTTTCTAGAGATAGATTGGTGCCTTCGGGAACATTGAGACAGGTGCT GCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTT GTCCTTAGTTACCAGCACGTAATGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGA AGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGT CGGTACAGAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGAT CGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGG TGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGC
the strains WS33, DT04 and DT06 have been deposited in China general microbiological culture Collection center (CGMCC) at 8/2/2021 with the preservation numbers of CGMCC No.22997, 22998 and 22999.
Example 2
WS33, DT04, and DT06 strains were inoculated into 250mL Erlenmeyer flasks containing 100mL enriched medium, respectively, and activated for 36h at 30 ℃ on a 180rpm constant temperature shaker. And after 36h, taking the activated bacteria liquid for plate counting, then taking a proper amount of activated bacteria liquid of the three strains based on the plate counting result, and compounding the activated bacteria liquid according to the proportion of 2:2:1 of the number of the cells to prepare the compound bacteria liquid. Inoculating the composite bacterial liquid into a denitrification culture medium according to the inoculation ratio of 1 percent, and then culturing for 4 days at constant temperature of 6 ℃ and 180 rpm. Inoculating the same volume of the high-temperature steam sterilized compound bacterial liquid as a control group, and culturing under the same other conditions as the inoculation group. After the culture period is finished, taking a water sample to measure the concentrations of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 1.
As can be seen from the measurement results in Table 1, after 4 days, the degradation rates of COD, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the treatment group inoculated with the composite bacterial liquid are 52.4%, 35.9%, 47.2% and 30.7% respectively; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are respectively only 0.5%, 0.1%, 0.2% and 0.2%. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is obvious under the low temperature condition of 6 ℃.
Table 1 shows the denitrification and COD removal effects of the complex bacteria at 6 DEG C
Example 3
WS33, DT04, and DT06 strains were inoculated into 250mL Erlenmeyer flasks containing 100mL enriched medium, respectively, and activated for 36h at 30 ℃ on a 180rpm constant temperature shaker. And after 36h, taking the activated bacteria liquid for plate counting, then taking a proper amount of activated bacteria liquid of the three strains based on the plate counting result, and compounding the activated bacteria liquid according to the proportion of 1:1:1 of the number of the cells to prepare the compound bacteria liquid. Inoculating the composite bacterial liquid into a denitrification culture medium according to the inoculation ratio of 1%, and then culturing for 4 days at the constant temperature of 8 ℃ and 180 rpm. Inoculating the same volume of the high-temperature steam sterilized compound bacterial liquid as a control group, and culturing under the same other conditions as the inoculation group. After the culture period is finished, taking a water sample to measure the concentrations of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 2.
As can be seen from the measurement results in Table 2, after 4 days, the degradation rates of COD, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the treatment group inoculated with the composite bacterial liquid are respectively 62.3%, 51.7%, 55.4% and 36.9%; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are only 1.2%, 0.6%, 0.2% and 0.1% respectively. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is improved to some extent under the low temperature condition of 8 ℃ compared with that under the temperature condition of 6 ℃, and the effect is obvious.
Table 2 shows the denitrification and COD removal effects of the complex bacteria at 8 DEG C
Example 4
The activated compound bacterial liquid in the example 2 is inoculated into a denitrification culture medium according to the inoculation ratio of 1 percent, and then is cultured for 4 days under the conditions of 10 ℃ and 180 rpm. And inoculating the compound bacterial liquid sterilized by high-temperature steam with the same volume as the inoculated group as a control group, wherein other culture conditions are the same as those of the inoculated group. And after the culture period is finished, taking a water sample to measure the concentrations of ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 3.
As can be seen from the measurement results in Table 3, after 4 days, the degradation rates of COD, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen of the treatment group inoculated with the composite bacterial liquid are 78.2%, 65.4%, 67.2% and 48.1% respectively; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are respectively only 0.5%, 0.1%, 0.2% and 0.2%. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is obvious under the condition of low temperature of 10 ℃.
Table 3 shows the denitrification and COD removal effects of the complex bacteria at 10 ℃
Example 5
WS33, DT04, and DT06 strains were inoculated into 250mL Erlenmeyer flasks containing 100mL enriched medium, respectively, and activated for 36h at 30 ℃ on a 180rpm constant temperature shaker. And after 36h, taking the activated bacterial liquid, carrying out plate counting, taking a proper amount of activated bacterial liquid of the three strains based on the plate counting result, and compounding according to the proportion of 3:5:2 of the number of the cells to prepare the compound bacterial liquid.
Inoculating the activated compound bacterial liquid into a denitrification culture medium according to the inoculation ratio of 1 percent, and then culturing for 4 days at constant temperature of 15 ℃ and 180 rpm. Inoculating the same volume of the high-temperature steam sterilized compound bacterial liquid as a control group, and culturing under the same other conditions as the inoculation group. And after the culture period is finished, taking a water sample to measure the concentrations of ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 4.
As can be seen from the results of the measurement in Table 4, the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the treatment group inoculated with the composite bacterial liquid after 4 days are 85.5%, 85.8%, 77.6% and 61.9%, respectively; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are respectively only 0.2%, 0.4% and 0.0%. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is obvious under the low temperature condition of 15 ℃.
Table 4 shows the denitrification and COD removal effects of the complex bacteria at 15 ℃
Example 6
WS33, DT04, and DT06 strains were inoculated into 250mL Erlenmeyer flasks containing 100mL enriched medium, respectively, and activated for 36h at 30 ℃ on a 180rpm constant temperature shaker. And after 36h, taking the activated bacteria liquid for plate counting, then taking a proper amount of activated bacteria liquid of the three strains based on the plate counting result, and compounding the activated bacteria liquid according to the proportion of 3:3:2 of the number of the cells to prepare the compound bacteria liquid.
Inoculating the activated compound bacterial liquid into a denitrification culture medium according to the inoculation ratio of 1.5%, and then culturing for 4 days at constant temperature of 25 ℃ and 180 rpm. Inoculating the same volume of the high-temperature steam sterilized compound bacterial liquid as a control group, and culturing under the same other conditions as the inoculation group. And after the culture period is finished, taking a water sample to measure the concentrations of ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 5.
As can be seen from the results of the measurement in Table 5, after 4 days, the degradation rates of COD, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the treatment group inoculated with the composite bacterial liquid are respectively 98.7%, 98.9%, 95.4% and 95.1%; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are respectively only 0.2%, 0.0%, 0.1% and 0.0%. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is obvious under the condition of 25 ℃.
Table 5 shows the denitrification and COD removal effects of the complex bacteria at 25 ℃
Example 7
The activated compound bacterial liquid in the example 6 is inoculated into a denitrification culture medium according to the inoculation ratio of 2 percent, and then is cultured for 4 days under the conditions of 40 ℃ and 180 rpm. Inoculating the same volume of the high-temperature steam sterilized compound bacterial liquid as a control group, and culturing under the same other conditions as the inoculation group. And after the culture period is finished, taking a water sample to measure the concentrations of ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen. The results are shown in Table 6.
As can be seen from the results of the measurement in Table 6, after 4 days, the degradation rates of COD, ammonia nitrogen, nitrate nitrogen and nitrite nitrogen in the treatment group inoculated with the composite bacterial liquid are respectively 99.4%, 99.2%, 98.8% and 92.3%; the degradation rates of COD, ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen in the water sample of the control group inoculated with the sterilized composite bacterial liquid are respectively only 0.1%, 0.2%, 0.0% and 0.0%. The result shows that the COD degradation and denitrification effect of the inoculated composite bacteria is obvious under the low temperature condition of 40 ℃. Table 5 shows the denitrification and COD removal effects of the complex bacteria at 40 ℃.
Table 6 shows the effect of denitrifying and removing COD at 40 deg.C
Sequence listing
<110> Shenyang landscape architecture shares Ltd
<120> low temperature resistant composite denitrifying bacteria and application thereof in sewage treatment
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1400
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
tggctcagat tgaacgctgg cggcaggcct aacacatgca agtcgagcgg cagcggaaag 60
tagcttgcta ctttgccggc gagcggcgga cgggtgagta atgtctggga aactgcctga 120
tggaggggga taactactgg aaacggtagc taataccgca tgacctcgaa agagcaaagt 180
gggggatctt cggacctcac gccatcggat gtgcccagat gggattagct agtaggtgag 240
gtaatggctc acctaggcga cgatccctag ctggtctgag aggatgacca gccacactgg 300
aactgagaca cggtccagac tcctacggga ggcagcagtg gggaatattg cacaatgggc 360
gcaagcctga tgcagccatg ccgcgtgtgt gaagaaggcc ttagggttgt aaagcacttt 420
cagcgaggag gaaggcatca tacttaatac gtgtggtgat tgacgttact cgcagaagaa 480
gcaccggcta actccgtgcc agcagccgcg gtaatacgga gggtgcaagc gttaatcgga 540
attactgggc gtaaagcgca cgcaggcggt ttgttaagtc agatgtgaaa tccccgcgct 600
taacgtggga actgcatttg aaactggcaa gctagagtct tgtagagggg ggtagaattc 660
caggtgtagc ggtgaaatgc gtagagatct ggaggaatac cggtggcgaa ggcggccccc 720
tggacaaaga ctgacgctca ggtgcgaaag cgtggggagc aaacaggatt agataccctg 780
gtagtccacg ctgtaaacga tgtcgacttg gaggttgtgc ccttgaggcg tggcttccgg 840
agctaacgcg ttaagtcgac cgcctgggga gtacggccgc aaggttaaaa ctcaaatgaa 900
ttgacggggg cccgcacaag cggtggagca tgtggtttaa ttcgatgcaa cgcgaagaac 960
cttacctact cttgacatcc acggaattcg ccagagatgg cttagtgcct tcgggaaccg 1020
tgagacaggt gctgcatggc tgtcgtcagc tcgtgttgtg aaatgttggg ttaagtcccg 1080
caacgagcgc aacccttatc ctttgttgcc agcacgtgat ggtgggaact caaaggagac 1140
tgccggtgat aaaccggagg aaggtgggga tgacgtcaag tcatcatggc ccttacgagt 1200
agggctacac acgtgctaca atggcatata caaagagaag cgaactcgcg agagcaagcg 1260
gacctcataa agtatgtcgt agtccggatt ggagtctgca actcgactcc atgaagtcgg 1320
aatcgctagt aatcgtagat cagaatgcta cggtgaatac gttcccgggc cttgtacaca 1380
ccgcccgtca caccatggga 1400
<210> 2
<211> 1407
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
cccgaaggtt agactagcta cttctggtgc aacccactcc catggtgtga cgggcggtgt 60
gtacaaggcc cgggaacgta ttcaccgcga cattctgatt cgcgattact agcgattccg 120
acttcacgca gtcgagttgc agactgcgat ccggactacg atcggttttc tgggattagc 180
tccacctcgc ggcttggcaa ccctctgtac cgaccattgt agcacgtgtg tagcccaggc 240
cgtaagggcc atgatgactt gacgtcatcc ccaccttcct ccggtttgtc accggcagtc 300
tccttagagt gcccaccatt acgtgctggt aactaaggac aagggttgcg ctcgttacgg 360
gacttaaccc aacatctcac gacacgagct gacgacagcc atgcagcacc tgtctcaatg 420
ttcccgaagg caccaatcta tctctagaaa gttcattgga tgtcaaggcc tggtaaggtt 480
cttcgcgttg cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 540
atttgagttt taaccttgcg gccgtactcc ccaggcggtc aacttaatgc gttagctgcg 600
ccactaagag ctcaaggctc ccaacggcta gttgacatcg tttacggcgt ggactaccag 660
ggtatctaat cctgtttgct ccccacgctt tcgcacctca gtgtcagtat cagtccaggt 720
ggtcgccttc gccactggtg ttccttccta tatctacgca tttcaccgct acacaggaaa 780
ttccaccacc ctctaccata ctctagctcg tcagttttga atgcagttcc caggttgagc 840
ccggggattt cacatccaac ttaacgaacc acctacgcgc gctttacgcc cagtaattcc 900
gattaacgct tgcaccctct gtattaccgc ggctgctggc acagagttag ccggtgctta 960
ttctgtcggt aacgtcaaaa ttgcagagta ttaatctaca acccttcctc ccaacttaaa 1020
gtgctttaca atccgaagac cttcttcaca cacgcggcat ggctggatca ggctttcgcc 1080
cattgtccaa tattccccac tgctgcctcc cgtaggagtc tggaccgtgt ctcagttcca 1140
gtgtgactga tcatcctctc agaccagtta cggatcgtcg ccttggtgag ccattacccc 1200
accaactagc taatccgacc taggctcatc tgatagcgca aggcccgaag gtcccctgct 1260
ttctcccgta ggacgtatgc ggtattagcg tccgtttccg aacgttatcc cccactacca 1320
ggcagattcc taggcattac tcacccgtcc gccgctctca agagaagcaa gcttctctct 1380
accgctcgac ttgcatgtgt tagtctg 1407
<210> 3
<211> 1400
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ctcagattga acgctggcgg caggcctaac acatgcaagt cgagcggtag agagaagctt 60
gcttctcttg agagcggcgg acgggtgagt aatgcctagg aatctgcctg gtagtggggg 120
ataacgttcg gaaacggacg ctaataccgc atacgtccta cgggagaaag caggggacct 180
tcgggccttg cgctatcaga tgagcctagg tcggattagc tagttggtga ggtaatggct 240
caccaaggcg acgatccgta actggtctga gaggatgatc agtcacactg gaactgagac 300
acggtccaga ctcctacggg aggcagcagt ggggaatatt ggacaatggg cgaaagcctg 360
atccagccat gccgcgtgtg tgaagaaggt cttcggattg taaagcactt taagttggga 420
ggaagggcag ttacctaata cgtgattgtt ttgacgttac cgacagaata agcaccggct 480
aactctgtgc cagcagccgc ggtaatacag agggtgcaag cgttaatcgg aattactggg 540
cgtaaagcgc gcgtaggtgg tttgttaagt tggatgtgaa atccccgggc tcaacctggg 600
aactgcattc aaaactgact gactagagta tggtagaggg tggtggaatt tcctgtgtag 660
cggtgaaatg cgtagatata ggaaggaaca ccagtggcga aggcgaccac ctggactaat 720
actgacactg aggtgcgaaa gcgtggggag caaacaggat tagataccct ggtagtccac 780
gccgtaaacg atgtcaacta gccgttggaa gccttgagct tttagtggcg cagctaacgc 840
attaagttga ccgcctgggg agtacggccg caaggttaaa actcaaatga attgacgggg 900
gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg 960
ccttgacatc caatgaactt tctagagata gattggtgcc ttcgggaaca ttgagacagg 1020
tgctgcatgg ctgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gtaacgagcg 1080
caacccttgt ccttagttac cagcacgtaa tggtgggcac tctaaggaga ctgccggtga 1140
caaaccggag gaaggtgggg atgacgtcaa gtcatcatgg cccttacggc ctgggctaca 1200
cacgtgctac aatggtcggt acagagggtt gccaagccgc gaggtggagc taatcccata 1260
aaaccgatcg tagtccggat cgcagtctgc aactcgactg cgtgaagtcg gaatcgctag 1320
taatcgcgaa tcagaatgtc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380
acaccatggg agtgggttgc 1400
Claims (10)
1. A low temperature resistant composite denitrifying bacteria is characterized in that: the denitrobacteria complex is a mixture of at least two strains of Rahnella aquatilis strain WS33, Pseudomonas sp strain DT04, and Pseudomonas sp strain DT 06.
2. The low temperature resistant denitrogenation complex of claim 1, wherein: the composite denitrobacteria are Rahnella aquatilis strain WS33, Pseudomonas (Pseudomonas sp.) strain DT04 and Pseudomonas (Pseudomonas sp.) strain DT 06.
3. The low temperature resistant denitrogenation complex of claim 2, wherein: the composite denitrifying bacteria are Rahnella aquatilis strain WS33, Pseudomonas (Pseudomonas sp.) strain DT04 and Pseudomonas (Pseudomonas sp.) strain DT06, and the number ratio of the strains is 2-5:2-8: 1-3.
4. The low temperature resistant denitrogenation complex as claimed in any one of claims 1 to 3, wherein: the Pseudomonas (Pseudomonas sp.) strain DT04, the Rahnella aquatilis (Rahnella aquatilis) strain WS33 and the Pseudomonas (Pseudomonas sp.) strain DT06 are all preserved in the China general microbiological culture Collection center (CGMCC) at 8 and 2 months in 2021, and the preservation numbers of the strains are CGMCC No.22997, 22998 and 22999 respectively.
5. The use of the low temperature resistant denitrifier according to claim 1 in sewage treatment.
6. The use of claim 5, wherein the low temperature resistant denitrifying bacteria are used in sewage treatment at 6-30 ℃.
7. The use of claim 6, wherein the low temperature resistant denitrogenating bacteria are activated by liquid culture medium respectively, and the activation culture conditions are as follows: shaking at 80-180rpm at 20-35 deg.C for 24-48 hr with initial pH of the culture medium of 7.0-8.0, mixing the activated strains, and adding into the polluted water to be treated.
8. Use according to claim 7, characterized in that: mixing the activated bacteria solutions, and mixing the mixed solution with a Rahnella aquatilis strain WS33, a Pseudomonas sp (Pseudomonas sp) strain DT04 and a Pseudomonas sp (Pseudomonas sp) strain DT06 according to the number ratio of the strains of 2-5:2-8: 1-3.
9. The use according to claim 5, wherein the form of inorganic nitrogen in said contaminated water body is ammoniacal nitrogen, nitrate nitrogen and nitrite nitrogen; wherein the concentration range of ammoniacal nitrogen is 10-300mg/L, the concentration range of nitrate nitrogen is 5-800mg/L, the concentration range of nitrite state is 0.2-100mg/L, and the concentration range of COD is 50-4000 mg/L.
10. Use according to claim 5, wherein the effluent has a salinity (in NaCl concentration) of 0.1-30g/L and a pH of 6.0-9.0.
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|---|---|---|---|---|
| CN101705202A (en) * | 2009-12-03 | 2010-05-12 | 华中农业大学 | Pseudomonas stutzeri YZN-001 for removing ammoniacal nitrogen, nitrate nitrogen and nitrous nitrogen in water body and application |
| CN102061276A (en) * | 2010-11-15 | 2011-05-18 | 北京大学 | Pseudomonas sp. strain for biological denitrification under low temperature and application thereof |
| US20210269341A1 (en) * | 2019-01-15 | 2021-09-02 | Agro-environmental Protection Institute, Ministry Of Agriculture And Rural Affairs | Las-degrading and/or n-removing bacterium and application thereof |
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| CN101705202A (en) * | 2009-12-03 | 2010-05-12 | 华中农业大学 | Pseudomonas stutzeri YZN-001 for removing ammoniacal nitrogen, nitrate nitrogen and nitrous nitrogen in water body and application |
| CN102061276A (en) * | 2010-11-15 | 2011-05-18 | 北京大学 | Pseudomonas sp. strain for biological denitrification under low temperature and application thereof |
| US20210269341A1 (en) * | 2019-01-15 | 2021-09-02 | Agro-environmental Protection Institute, Ministry Of Agriculture And Rural Affairs | Las-degrading and/or n-removing bacterium and application thereof |
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