CN116574639B - Biological deodorization composite microbial inoculant and preparation method and application thereof - Google Patents
Biological deodorization composite microbial inoculant and preparation method and application thereof Download PDFInfo
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- CN116574639B CN116574639B CN202310355598.5A CN202310355598A CN116574639B CN 116574639 B CN116574639 B CN 116574639B CN 202310355598 A CN202310355598 A CN 202310355598A CN 116574639 B CN116574639 B CN 116574639B
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- 238000004332 deodorization Methods 0.000 title claims abstract description 81
- 239000002131 composite material Substances 0.000 title claims abstract description 56
- 230000000813 microbial effect Effects 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 239000002054 inoculum Substances 0.000 title abstract description 13
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 111
- 230000001580 bacterial effect Effects 0.000 claims abstract description 110
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 60
- 239000007788 liquid Substances 0.000 claims abstract description 56
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 50
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 50
- 241000186359 Mycobacterium Species 0.000 claims abstract description 40
- 239000000945 filler Substances 0.000 claims description 79
- 238000000034 method Methods 0.000 claims description 28
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 24
- 229910052799 carbon Inorganic materials 0.000 claims description 24
- 238000011081 inoculation Methods 0.000 claims description 21
- 238000000855 fermentation Methods 0.000 claims description 20
- 230000004151 fermentation Effects 0.000 claims description 20
- 238000012258 culturing Methods 0.000 claims description 19
- 241000605272 Acidithiobacillus thiooxidans Species 0.000 claims description 11
- 238000005507 spraying Methods 0.000 claims description 8
- 230000005660 hydrophilic surface Effects 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 241000187488 Mycobacterium sp. Species 0.000 claims 1
- 241001290773 Acidiphilium acidophilum Species 0.000 abstract description 23
- 230000002159 abnormal effect Effects 0.000 abstract description 6
- 238000001914 filtration Methods 0.000 abstract description 5
- 239000001963 growth medium Substances 0.000 description 46
- 239000000243 solution Substances 0.000 description 39
- 238000009630 liquid culture Methods 0.000 description 26
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 22
- 241000894006 Bacteria Species 0.000 description 20
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- 230000001877 deodorizing effect Effects 0.000 description 9
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 9
- 241000266272 Acidithiobacillus Species 0.000 description 8
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 8
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- 238000004659 sterilization and disinfection Methods 0.000 description 6
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- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
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- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 4
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- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- KKUKTXOBAWVSHC-UHFFFAOYSA-N Dimethylphosphate Chemical compound COP(O)(=O)OC KKUKTXOBAWVSHC-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 229920002472 Starch Polymers 0.000 description 2
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- MVIHXDVCXNPIAL-UHFFFAOYSA-N [4-[3-oxo-1-(4-sulfooxyphenyl)-2-benzofuran-1-yl]phenyl] hydrogen sulfate Chemical compound C1=CC(OS(=O)(=O)O)=CC=C1C1(C=2C=CC(OS(O)(=O)=O)=CC=2)C2=CC=CC=C2C(=O)O1 MVIHXDVCXNPIAL-UHFFFAOYSA-N 0.000 description 2
- 238000011481 absorbance measurement Methods 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Chemical compound C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 2
- TZCXTZWJZNENPQ-UHFFFAOYSA-L barium sulfate Chemical compound [Ba+2].[O-]S([O-])(=O)=O TZCXTZWJZNENPQ-UHFFFAOYSA-L 0.000 description 2
- 239000001110 calcium chloride Substances 0.000 description 2
- 229910001628 calcium chloride Inorganic materials 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 description 2
- 238000005260 corrosion Methods 0.000 description 2
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- 239000012895 dilution Substances 0.000 description 2
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 2
- 239000011790 ferrous sulphate Substances 0.000 description 2
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- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 2
- 235000013923 monosodium glutamate Nutrition 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
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- YNPNZTXNASCQKK-UHFFFAOYSA-N phenanthrene Chemical compound C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 229940073490 sodium glutamate Drugs 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- -1 sulfide ions Chemical class 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000001651 autotrophic effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 229910001422 barium ion Inorganic materials 0.000 description 1
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- 239000001045 blue dye Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
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- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000002781 deodorant agent Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012954 diazonium Substances 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-O diazynium Chemical compound [NH+]#N IJGRMHOSHXDMSA-UHFFFAOYSA-O 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
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- 229910001447 ferric ion Inorganic materials 0.000 description 1
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- 238000012165 high-throughput sequencing Methods 0.000 description 1
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- 150000002632 lipids Chemical class 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical compound [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 1
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 1
- 229960000907 methylthioninium chloride Drugs 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
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- 230000001502 supplementing effect Effects 0.000 description 1
- HPQYKCJIWQFJMS-UHFFFAOYSA-L tetrathionate(2-) Chemical compound [O-]S(=O)(=O)SSS([O-])(=O)=O HPQYKCJIWQFJMS-UHFFFAOYSA-L 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 241001446247 uncultured actinomycete Species 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
- B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
- B01D53/84—Biological processes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D2257/00—Components to be removed
- B01D2257/90—Odorous compounds not provided for in groups B01D2257/00 - B01D2257/708
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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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- 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
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass 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
- C12R2001/32—Mycobacterium
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
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- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Environmental & Geological Engineering (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
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- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
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- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a biological deodorization composite microbial inoculant, which is prepared from a first microbial inoculant and a second microbial inoculant according to the volume ratio of (1-5): (3-10); wherein the first microbial inoculum comprises bacillus subtilis and thiobacillus acidophilus, and the concentration of the microbial inoculum is 1 multiplied by 10 6~1×109 CFU/mL; the second bacterial agent comprises mycobacterium, and the bacterial liquid concentration is 1X 10 4~1×106 CFU/mL. The invention also discloses a preparation method and application of the biological deodorization composite bacterial agent. The biological deodorization composite microbial inoculum can effectively solve the problems of long domestication and debugging starting period of a biological trickling filtration tower, poor capability of resisting abnormal working conditions and poor capability of resisting severe environments.
Description
Technical Field
The invention relates to the technical field of environmental protection and treatment, in particular to a biological deodorization composite microbial inoculant, a preparation method and application thereof.
Background
In the technical field of odor treatment, the biotrickling filter is widely used because of the advantages of low cost, low energy consumption, convenient operation and management and the like. The principle of the biotrickling filter for removing malodorous pollutants is as follows: the malodorous pollutant in the malodor is captured and utilized by the microorganism attached to the filler surface of the biotrickling filter, part of the malodorous pollutant is converted into small molecular substances such as CO 2、H2 O, and the other small molecular substances are converted into energy for the microorganism to grow and reproduce, so that the purpose of removing the malodorous pollutant in the malodor is achieved.
The removal efficiency of the biotrickling filter depends on the degradation and utilization capacity of microorganisms on malodorous pollutants.
The strain sources of the current biotrickling filter are mostly naturally domesticated, and the problem of long debugging period exists due to long microorganism generation period and long domestication period; meanwhile, the single odor pollution component easily causes single structure of domesticated flora, low biodiversity and poor stability, and causes poor capability of the biological trickling filter to resist abnormal working conditions (such as water cut, power failure, long-time overhaul and the like) or severe environments such as high temperature, cold, freezing and the like.
Therefore, a need exists to provide a biological deodorization composite microbial inoculum so as to solve the problems of long period of domestication and debugging and starting of a biological trickling filtration tower, poor capability of resisting abnormal working conditions and poor capability of resisting severe environments.
Disclosure of Invention
The invention aims to provide a biological deodorization composite microbial inoculum and a preparation method and application thereof, so as to solve the problems of long starting period, poor stability and the like of a biological filter.
In one aspect, the invention provides a biological deodorization composite microbial inoculant, which is prepared from a first microbial inoculant and a second microbial inoculant according to the volume ratio of (1-5): (3-10);
Wherein the first microbial inoculum comprises bacillus subtilis and thiobacillus acidophilus, and the concentration of the microbial inoculum is 1 multiplied by 10 6~1×109 CFU/mL; the second microbial inoculum comprises mycobacterium, and the concentration of the microbial inoculum is 1X 10 4~1×106 CFU/mL.
Optionally, the volume ratio of the first microbial agent to the second microbial agent is (1-2): (5-10), preferably 2:5.
Optionally, the bacterial liquid concentration of the first bacterial agent is 1×10 6~1×108 CFU/mL, preferably 1×10 7 CFU/mL.
Optionally, the concentration of the bacterial liquid of the second bacterial agent is 5×10 4~6×105 CFU/mL, preferably 1×10 5 CFU/mL.
Optionally, in the first microbial inoculum, the colony ratio of bacillus subtilis and thiobacillus acidophilus is (7-8): (1-5), preferably (7-8): (2-3).
On the other hand, the invention provides a preparation method of the biological deodorization composite bacterial agent, which comprises the following steps:
(1) Respectively carrying out fermentation culture to obtain a mycobacterium culture solution, a bacillus subtilis culture solution and a thiobacillus acidophilus culture solution;
(2) Mixing bacillus subtilis culture solution and thiobacillus acidophilus culture solution, adding sterilized carbonaceous filler into the obtained mixed bacterial solution, and culturing to obtain a first microbial inoculum;
(3) And adding the sterilized carbonaceous filler into a mycobacterium culture solution, and culturing to obtain a second microbial inoculum.
Optionally, in the step (2), the volume percentages of the bacillus subtilis culture solution and the thiobacillus acidophilus culture solution are respectively 70-80% and 20-30%, the sum of the bacillus subtilis culture solution and the thiobacillus acidophilus culture solution is 100%, and the adding proportion of the carbonaceous filler is 50-100 g of the carbonaceous filler per liter of the mixed bacterial solution; in the step (3), the adding proportion of the carbonaceous filler is 50-100 g of the carbonaceous filler per liter of the mycobacterium culture solution.
Optionally, the diameter of the carbonaceous filler is 5-10 mm, the specific surface area is not less than 250m 2/g, and the surface is hydrophilic.
On the other hand, the invention provides a use method of the biological deodorization composite bacterial agent, which sequentially comprises the following steps:
Step one, inoculating a first microbial inoculum to the lower half layer of the biological filter, which is close to the air inlet side, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor;
Step two, inoculating a second microbial inoculum to the upper half layer of the biological filter far away from the air inlet side, wherein the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor;
preferably, the use method sequentially comprises the following steps:
dispersing a carbon filler in a first microbial inoculum on the lower half layer of the biological filter near the air inlet side, spraying bacterial liquid in the first microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 1-2 days;
Dispersing the carbon filler in the second microbial inoculum on the upper half layer of the biological filter far away from the air inlet side, spraying bacterial liquid in the second microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 3 to 4 days.
On the other hand, the invention provides a use method of the biological deodorization composite bacterial agent, which sequentially comprises the following steps:
Step one, inoculating a first microbial inoculum to the upper half layer of the biological filter, which is close to the air inlet side, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor;
Step two, inoculating a second microbial inoculum to the lower half layer of the biological filter far away from the air inlet side, wherein the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor;
preferably, the use method sequentially comprises the following steps:
Dispersing a carbon filler in a first microbial inoculum on the upper half layer of the biological filter near the air inlet side, spraying bacterial liquid in the first microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 1-2 days;
dispersing the carbon filler in the second microbial inoculum on the lower half layer of the biological filter far away from the air inlet side, spraying bacterial liquid in the second microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 3 to 4 days.
According to the technical scheme, the biological deodorization composite microbial inoculant and the preparation method and application thereof have at least the following beneficial effects:
the biological deodorization composite microbial inoculum provided by the invention has good biocompatibility with carbon filler, and can be easily used as an dominant strain after inoculation.
Compared with a natural domesticated biological filter, the biological filter inoculated with the biological deodorization composite bacterial agent can finish domestication and starting rapidly, and the deodorization efficiency can be improved by 20% -40%.
The inoculation of the biological deodorization composite microbial inoculum can obviously shorten the debugging period of the biological filter, and enhance the resistance of the biological filter to severe environments such as cold, hot and the like and abnormal working conditions such as long-time idling, water cut, power failure, overhaul and the like.
Drawings
Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. In the drawings:
FIG. 1 is a graph showing the concentration of elemental sulfur during the degradation of elemental sulfur by 3 pure bacteria selected in example 1, namely, mycobacterium, bacillus subtilis and Acidithiobacillus;
FIG. 2 is a graph showing the change in sulfate ion concentration during the degradation of elemental sulfur by 3 pure bacteria selected in example 1, namely Mycobacterium, bacillus subtilis and Acidithiobacillus;
FIG. 3 shows growth curves of 3 pure bacteria obtained by screening in example 1;
FIG. 4 is a graph showing the comparison of the hydrogen sulfide removal of the biological filter inoculated with the biological deodorization composite bacterial agent and the natural domestication tower in example 1;
FIG. 5 is a graph showing the comparison of the biological filter inoculated with the biological deodorization composite bacterial agent and the natural domestication tower in example 1 against the influence of severe winter weather.
Detailed Description
The present invention will be described in detail with reference to the following embodiments for a full understanding of the objects, features and effects of the present invention, but the present invention is not limited thereto. The present invention will be described in detail with reference to the following embodiments for a full understanding of the objects, features, and effects of the present invention. The process of the present invention is carried out by methods or apparatus conventional in the art, except as described below.
In a first aspect, the invention provides a biological deodorization composite microbial inoculant which can be applied to a biological filter. The biological deodorization composite microbial inoculum consists of a first microbial inoculum and a second microbial inoculum. The volume ratio of the first microbial inoculum to the second microbial inoculum is (1-5): (3-10), preferably (1-2): (5-10), more preferably 2:5.
The deodorizing bacteria in the first microbial inoculum are bacillus subtilis (Bacillus subtilis) and thiobacillus acidophilus (Sulfobacillus), and the colony ratio of the bacillus subtilis and the thiobacillus acidophilus is (7-8): (1-5), preferably (7-8): (2-3). The concentration of the bacterial liquid in the first bacterial agent is 1X 10 6~1×109 CFU/mL, preferably 1X 10 6~1×108 CFU/mL, and most preferably 1X 10 7 CFU/mL.
The bacillus subtilis in the first microbial inoculum is an aerobic bacterium, and organic matters, elemental sulfur and tetrathionate are taken as electron donors to carry out growth metabolism. The bacillus subtilis has good acid resistance and heat resistance, the survival pH value ranges from 2.0 to 6.0, the temperature ranges from 20 ℃ to 70 ℃, spores are generated to become dormancy when the bacillus subtilis encounters a severe environment unsuitable for survival, and vegetative cells are regenerated when the environment is suitable. The spores can resist drought, high heat, high acid and alkali and hypertonic environment.
The thiobacillus acidophilus in the first microbial inoculum is an aerobic bacilliform bacterium and has acidophilic property. The method can take ferrous iron, elemental sulfur and sulfide as raw materials for autotrophic growth, and in the sulfur oxidation process, hydrogen sulfide, elemental sulfur and sulfite are gradually oxidized and degraded.
The deodorant bacteria in the second microbial agent is Mycobacterium (Mycobacterium). The concentration of the second microbial inoculum is 1X 10 4~1×106 CFU/mL, preferably 5X 10 4~6×105 CFU/mL, and most preferably 1X 10 5 CFU/mL.
The mycobacteria in the second agent is an aerobic actinomycete, the cell wall of which contains a large amount of lipids, also known as acid fast bacilli, which are resistant to strong acid environments. The mycobacterium can utilize various carbon sources, and can mineralize and degrade polycyclic aromatic hydrocarbon such as anthracene, phenanthrene and the like under the condition of supplementing a small amount of organic nutrients. In addition, mycobacteria produce arylsulfonases that degrade sulfate functionality in organic malodorous molecules into free phenolphthalein and residual salts.
The inventor finds that the bacillus subtilis, the thiobacillus acidophilus and the mycobacterium are prepared into the composite microbial inoculum according to the concentration and the proportion, and the bacteria can be matched with each other well on the premise of playing the respective functional characteristics, so that the effect of degrading the sulfur simple substance is achieved well.
The biological deodorizing composite bacterial agent can be obtained by separating and purifying in a laboratory and preserving the bacterial strains, and can also be commercial bacterial strains obtained through market purchase.
In a second aspect, the invention provides a preparation method of a biological deodorization composite bacterial agent, which comprises the following steps:
(1) Respectively fermenting and culturing to obtain mycobacterium culture solution, bacillus subtilis culture solution and thiobacillus acidophilus culture solution.
Mycobacteria can be cultured using modified Gaoshan No. 1 liquid medium. The formula of the improved Gaoshi No. 1 liquid culture medium is as follows: 1g/L of potassium nitrate, 0.5g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate, 0.01g/L of ferrous sulfate, 0.5g/L of sodium chloride and 20g/L of soluble starch. As an example, mycobacteria can be fermented as follows: the mycobacterium is transferred to the modified Gaoshi No. 1 liquid culture medium for 14 times so as to carry out enrichment culture, and then the enriched culture bacterial liquid is inoculated into a fermentation tank filled with the modified Gaoshi No. 1 liquid culture medium according to the inoculation amount of 5 percent, and the mycobacterium culture liquid is obtained after the mycobacterium culture is cultured for 0.5 days in the environment with the stirring speed of 190r/min and the temperature of 30 ℃. Of course, the fermentation culture of Mycobacteria can also be referred to other known protocols in the art.
The bacillus subtilis can be cultured by adopting a modified Gao's No. 2 liquid culture medium. The formula of the improved Gaoshi No. 2 liquid culture medium is as follows: 42g/L glucose, 14g/L sodium glutamate, 0.5g/L magnesium sulfate heptahydrate, 0.5g/L potassium chloride, 1g/L potassium dihydrogen phosphate, 0.15mg/L ferrous sulfate heptahydrate, 5mg/L manganese sulfate hydrate, 0.16mg/L copper sulfate pentahydrate, pH7.0. As an example, bacillus subtilis may be fermented as follows: and transferring the bacillus subtilis in the modified Gao's No. 2 liquid culture medium for 14 times so as to perform enrichment culture, inoculating the enriched culture bacterial liquid into a fermentation tank filled with the modified Gao's No. 2 liquid culture medium according to an inoculum size of 5%, and culturing for 0.5 days in an environment with a stirring speed of 180r/min and a temperature of 32 ℃ to obtain the bacillus subtilis culture liquid. Of course, the fermentation culture of Bacillus subtilis can also be referred to other known schemes in the art.
The Acidithiobacillus may be cultured in 9K liquid culture medium. The formula of the 9K liquid culture medium is as follows: 3g/L of ammonium sulfate, 0.5g/L of dimethyl hydrogen phosphate, 0.5g/L of magnesium sulfate heptahydrate, 0.1g/L of potassium chloride, 6.8mg/L of calcium chloride, 6.2mg/L of potassium nitrate, 5g/L of elemental sulfur and 0.2g/L of yeast extract. As an example, the Acidithiobacillus may be subjected to fermentation culture by the following method: the thiobacillus acidophilus is transferred to a 9K liquid culture medium for 14 times so as to carry out enrichment culture, then the bacteria liquid of the enrichment culture is inoculated to a fermentation tank filled with the 9K liquid culture medium according to the inoculation amount of 5 percent, and the bacteria liquid is cultured for 4 days in the environment with the stirring speed of 190r/min and the temperature of 40 ℃ so as to obtain the thiobacillus acidophilus culture liquid. Of course, the fermentation culture of thiobacillus acidophilus can also be referred to other known schemes in the art.
(2) And mixing the bacillus subtilis culture solution and the thiobacillus acidophilus culture solution, adding sterilized carbonaceous filler into the obtained mixed bacterial solution, and culturing to obtain the first microbial inoculum.
The added volume ratio of the bacillus subtilis culture solution and the acidophilic thiobacillus culture solution is 70-80% and 20-30%, respectively, and the sum of the two is 100%.
The carbon filler has the diameter of 5-10 mm, the specific surface area of >250m 2/g, good moisture retention and air permeability, hydrophilic surface, acid corrosion resistance, stable structure and difficult hardening. The sterilization of the carbonaceous filler may be autoclaving. The adding proportion of the carbonaceous filler is that 50-100 g of the carbonaceous filler is added per liter of mixed bacterial liquid.
The culture conditions are preferably the optimal culture conditions of bacillus subtilis and thiobacillus acidophilus until the concentration of the bacterial liquid is 1X 10 6~1×109 CFU/mL.
As a preferred example, the carbonaceous filler is sterilized by using a pressure of 0.1MPa, namely 103.4kPa (1.05 kg/cm 2) under a steam pressure, the temperature in the sterilizing pot reaches 121 ℃, the conditions of 15-30 min are maintained, then the carbonaceous filler is added into the mixed bacterial liquid (75% of bacillus subtilis and 25% of thiobacillus acidophilus) according to 50g/L, and the culture is carried out for about 2 days in an environment with a stirring speed of 190r/min and a temperature of 35 ℃ until the bacterial liquid concentration in the culture liquid reaches 1X 10 7 cfu/mL, so as to obtain the first bacterial agent.
(3) And adding the sterilized carbonaceous filler into a mycobacterium culture solution, and culturing to obtain a second microbial inoculum.
The carbon filler has the diameter of 5-10 mm, the specific surface area of >250m 2/g, good moisture retention and air permeability, hydrophilic surface, acid corrosion resistance, stable structure and difficult hardening. The sterilization of the carbonaceous filler may be autoclaving. The adding proportion of the carbonaceous filler is that 50-100 g of the carbonaceous filler is added per liter of the mycobacterium culture solution.
The culture conditions are preferably the optimal culture conditions for Mycobacteria until the concentration of the bacterial liquid is 1X 10 4~1×106 CFU/mL.
As a preferred example, the carbonaceous filler is sterilized by using a pressure of 0.1MPa, i.e., a steam pressure of 103.4kPa (1.05 kg/cm 2), the temperature in the sterilizing pot is up to 121℃and maintained for 15 to 30 minutes, then the carbonaceous filler is added to the Mycobacterium broth at 50g/L, and the culture is carried out for about 1 day in an environment with a stirring speed of 190r/min and a temperature of 30℃until the concentration of the broth in the broth is up to 1X 10 5 cfu/mL, thereby obtaining the second microbial inoculum.
It should be noted that the sequence of the steps (2) and (3) is not limited in the present invention, and the steps (2) and (3) may be performed simultaneously, and the step (2) is performed before the step (3) is performed, or the step (3) is performed before the step (2) is performed, so long as the subsequent use is not affected.
The biological deodorization composite microbial inoculum can be applied to various biological filters, and the structure of the biological filter can refer to the related scheme in the prior art.
It should be noted that, when the biological deodorization composite microbial agent is specifically used, the specific inoculation positions of the first microbial agent and the second microbial agent need to be determined by combining the positions of the air inlets of the biological filter, and the basic principle is that the first microbial agent should be inoculated on a half-layer of filler close to the air inlets, and the second microbial agent should be inoculated on a half-layer of filler far away from the air inlets.
For the biological filter with the air inlet arranged at the lower part, the application method of the biological deodorization composite bacterial agent sequentially comprises the following steps:
Step one: and inoculating the first microbial inoculum to the lower half layer of the biological filter close to the air inlet side, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor.
Specifically, the carbonaceous filler in the first microbial inoculum is uniformly dispersed in the lower half layer of the biological filter, bacterial liquid in the first microbial inoculum is uniformly sprayed on the surface of the carbonaceous filler, and the biological filter is started to operate for 1 to 2 days.
And step two, inoculating a second microbial inoculum to the upper half layer of the filler of the biological filter far away from the air inlet side, wherein the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor.
Specifically, the carbonaceous filler in the second microbial inoculum is uniformly dispersed on the upper half layer of the filler of the biological filter far away from the air inlet, the bacterial liquid in the second microbial inoculum is uniformly sprayed on the surface of the carbonaceous filler, the biological filter is started to operate for 3 to 4 days, and the outlet can reach a stable state.
The application method of the biological deodorization composite bacterial agent comprises the following steps of:
step one, inoculating a first microbial inoculum to the upper half layer of the biological filter, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor.
Specifically, the carbonaceous filler in the first microbial inoculum is uniformly dispersed on the upper half layer of the biological filter, bacterial liquid in the first microbial inoculum is uniformly sprayed on the surface of the carbonaceous filler, and the biological filter is started to operate for 1 to 2 days.
And step two, inoculating a second microbial inoculum to the lower half layer of the filler of the biological filter, wherein the specific method is to spray the microbial inoculum on the surface of the filler through a peripheral window, and the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor.
Specifically, the carbonaceous filler in the second microbial inoculum is uniformly dispersed in the lower half layer of the filler of the biological filter, bacterial liquid in the second microbial inoculum is uniformly sprayed on the surface of the carbonaceous filler, the biological filter is started to operate for 3 to 4 days, and the outlet can reach a stable state.
According to the high-throughput sequencing result of the 16S rRNA genes on the flora on the packing layers with different heights of the biological deodorization tower with stable operation, the abundance of bacillus subtilis and thiobacillus acidophilus in the first microbial inoculum is obviously higher than that of the half packing on the side close to the air inlet, and the abundance of the mycobacterium in the second microbial inoculum is opposite. Based on the above, when the biological deodorization composite bacterial agent is used, the first bacterial agent is inoculated to the half layer of the biological filter close to the air inlet, and the second bacterial agent is inoculated to the half layer of the biological filter far from the air inlet.
Examples
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
The formulation of the medium used in the following examples is as follows:
Modified Gaoshi No. 1 liquid medium: 1g/L of potassium nitrate, 0.5g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate, 0.01g/L of ferrous sulfate, 0.5g/L of sodium chloride, 20g/L of soluble starch and pH7.0. Modified Gaoshi No. 1 solid medium agar 15g/L was added to the modified Gaoshi No. 1 liquid medium.
The modified Gaoshi No. 2 liquid medium includes: 42g/L glucose, 14g/L sodium glutamate, 0.5g/L magnesium sulfate heptahydrate, 0.5g/L potassium chloride, 1g/L potassium dihydrogen phosphate, 0.15mg/L ferrous sulfate heptahydrate, 5mg/L manganese sulfate hydrate, 0.16mg/L copper sulfate pentahydrate, pH7.0. The solid culture medium No. 2 is added with 15g/L agar on the basis of the modified liquid culture medium No. 2 Gaoshi.
The 9K liquid medium includes: 3g/L of ammonium sulfate, 0.5g/L of dimethyl hydrogen phosphate, 0.5g/L of magnesium sulfate heptahydrate, 0.1g/L of potassium chloride, 6.8mg/L of calcium chloride, 6.2mg/L of potassium nitrate, 5g/L of elemental sulfur, 0.2g/L of yeast extract and pH2.0.9K solid Medium agar 15g/L was added on the basis of 9K liquid Medium.
Example 1
Culture and identification of strains (one)
(1) Preparation of initial seed suspension sample
1G of carbon filler is respectively taken on the upper part, the middle part and the bottom of a biological trickling filtration tower bed layer with good running state, and is marked as filler A, filler B and filler C in sequence, the biological trickling filtration tower bed layer is soaked in sterile physiological saline and then is placed in an ultrasonic cleaner for ultrasonic vibration for 10 minutes, and after the large carbon filler is removed by filtration, suspension A, suspension B and suspension C which are rich in deodorizing bacteria are prepared.
(2) Enrichment of bacterial species
And (3) respectively taking the supernatant in the suspension A, B, C in the step (1), and coating the supernatant in the modified Gaoshi No. 1 culture medium, the modified Gaoshi No. 2 culture medium and the 9K culture medium for 3-5d under the culture condition of 30 ℃ and 150r/min. And transferring 5mL of the liquid culture medium into a fresh modified Gaoshi No. 1 culture medium, a modified Gaoshi No. 2 culture medium and a 9K culture medium after 3-5d, culturing for 3-5d under the same culture condition, and transferring for 14 times to obtain enriched bacterial liquid.
(3) Preliminary screening of strains
Inoculating the enriched bacterial liquid which is subjected to 14 times of transfer in the step (1) of 0.1 mL onto a solid culture medium flat plate, uniformly coating by using a triangular coating rod, culturing in a constant temperature incubator for 2 days, picking differential bacterial colony lines from the flat plate, separating the lines twice, obtaining pure bacterial colonies, and numbering.
(4) Compound screening of bacterial
The pure colony in the step (3) is picked and cultured in a liquid culture medium for 48 hours to prepare bacterial suspension with OD 600 value of 0.8 (a certain amount of bacterial liquid is taken for centrifugal separation, and is washed 2 times by 0.9 percent physiological saline, and then the bacterial suspension with OD 600 is prepared by the physiological saline). Transferring 5mL of bacterial suspension into a liquid culture medium containing 5g/L of elemental sulfur (5 g/L elemental sulfur is added to a modified Gaoshi No. 1 culture medium and a modified Gaoshi No. 2 culture medium respectively, 5g/L elemental sulfur is not added to a 9K liquid culture medium additionally), transferring 5mL of sterilized normal saline into the liquid culture medium containing 5g/L elemental sulfur, and culturing 4 groups of culture mediums in a culture box at 30 ℃ for 14 days at 150r/min, wherein the elemental sulfur concentration and sulfate ion concentration are measured every day.
(5) Identification of species
The enriched and purified bacterial liquid is entrusted to Shanghai Maipu biotechnology Co., ltd for identification, pure bacteria enriched and grown in the modified Gao's No. 1 culture medium are identified as Mycobacterium (Mycobacterium), pure bacteria enriched and grown in the modified Gao's No. 2 culture medium are identified as bacillus subtilis (Bacillus subtilis), and pure bacterial colony enriched and grown in the 9K culture medium is identified as thiobacillus acidophilus (Sulfobacillus).
3 Strains of pure bacteria obtained by screening are stored in a laboratory refrigerator.
(II) Performance detection
(1) Simple substance sulfur removal experiment
The method for measuring the concentration of the elemental sulfur comprises the following steps: the methylene spectrophotometry is based on the principle that sulfide ions and para-amino dimethylaniline generate methylene blue dye in acid solution containing ferric ions, and absorbance measurement is carried out at the wavelength of 665nm, wherein the blue color is in direct proportion to the content of sulfide ions in a water sample.
Sulfate ion concentration determination method: the turbidimetry is based on the principle that sulfate in water reacts with barium ions in an acidic medium to generate barium sulfate suspension crystals, so that the solution is turbid, absorbance measurement is carried out at the wavelength of 410nm, and the turbidity degree is proportional to the concentration of sulfate ions.
The degradation effect of the modified Gao No.1 culture medium, the modified Gao No.2 culture medium and the mycobacterium, the bacillus subtilis and the thiobacillus acidophilus enriched in the 9K culture medium in the first part of the step (4) on the elemental sulfur with the initial concentration of 5g/L is shown in the figures 1 and 2. Wherein, fig. 1 is a change curve of the concentration of the elemental sulfur in the process of degrading the elemental sulfur by 3 pure bacteria, and fig. 2 is a change curve of the concentration of sulfate ions in the process of degrading the elemental sulfur by 3 pure bacteria.
As can be seen from FIG. 1, on day 4, the removal efficiencies of Mycobacterium, bacillus subtilis and Acidithiobacillus were 62%, 70% and 100% for elemental sulfur at an initial concentration of 5g/L, respectively.
(2) Optimal fermentation conditions
The optimal technological parameters of the fermentation tank culture of 3 pure bacteria are researched by a controlled variable method:
The optimal technological parameters of the mycobacteria fermentation tank culture are stirring speed 190r/min, pH value 6, temperature 30 ℃ and optimal culture time 1 day;
The optimal technological parameters of the bacillus subtilis fermentation tank culture are that the stirring speed is 180r/min, the pH value is 6.5, the temperature is 32 ℃, and the optimal culture time is 1 day;
The optimal technological parameters of the culture of the acidophilic thiobacillus fermenter are the stirring speed of 190r/min, the pH value of 2.0, the temperature of 40 ℃ and the optimal culture time of 5 days.
(3) Growth curve
The method comprises the following steps:
Step one, respectively measuring OD 600 values corresponding to different times of mycobacterium, bacillus subtilis and thiobacillus acidophilus in an improved Gao's No. 1 liquid culture medium, an improved Gao's No. 2 liquid culture medium and a 9K liquid culture medium;
Preparing 3 different bacterial suspensions with OD 600 value determined in the first step into diluted bacterial suspension with concentration gradient of 10 -1 by using physiological saline, selecting bacterial suspension with proper dilution factor, inoculating 0.1. 0.1 mL on a solid culture medium plate, uniformly coating by using a triangular coating rod, culturing in a constant temperature incubator (stirring speed 190r/min and temperature 32 ℃) for 2 days, selecting plate count capable of separating single bacterial colony, and calculating the bacterial quantity contained in the original bacterial suspension according to the dilution factor and unit cfu/mL.
The growth curves of the 3 pure bacteria of Mycobacterium, bacillus subtilis and Acidithiobacillus are shown in figure 3.
By calculation, the number of Mycobacterium, bacillus subtilis and Acidithiobacillus was 4.6X10. 10 4CFU/mL、3.8×106CFU/mL、7.3×107 CFU/mL, respectively, at OD 600 of the bacterial suspension.
(4) Mycobacterium secretion arylsulfonase assay
The presence of arylsulfonases was judged by the following experiment: a modified Gaoshi No. 1 liquid medium was inoculated with a Mycobacterium suspension having a 5mLOD 600 of 0.5, and after 2d incubation at a shaking speed of 190r/min and a temperature of 30℃5ml of a 0.1mol/L sodium carbonate solution and 5ml of a 0.2mol/L tripotassium phenolphthalein disulfate trihydrate (CAS No. 62625-16-5), the solution was observed to change from colorless to red after shaking uniformly. This is because mycobacterial secreted arylsulfonases can degrade sulfate molecules in tripotassium phenolphthalein disulfate trihydrate to free phenolphthalein and residual salts, and the added sodium carbonate reacts with phenolphthalein and produces a readily visible red diazonium reaction.
Preparation of biological deodorization composite bacterial agent
(1) Preparation of a Single Strain
The method comprises the steps of putting an improved Gaoshi No. 1 liquid culture medium into a fermentation tank A, inoculating 5% of a bacterial liquid rich in mycobacterium, which is subjected to primary screening after 14 times of transfer of the improved Gaoshi No. 1 liquid culture medium, and culturing for 0.5d in an environment with the stirring speed of 190r/min and the temperature of 30 ℃ to obtain a mycobacterium culture solution.
The method comprises the steps of placing an improved Gao's No. 2 liquid culture medium in a fermentation tank B, inoculating 5% of bacillus subtilis-enriched bacterial liquid which is subjected to primary screening after 14 times of transfer of the improved Gao's No. 2 liquid culture medium, and culturing for 0.5d in an environment with the stirring speed of 180r/min and the temperature of 32 ℃ to obtain a bacillus subtilis culture liquid.
And (3) placing a 9K liquid culture medium in a fermentation tank C, inoculating 5% of bacterial liquid which is subjected to primary screening after 14 times of 9K liquid culture medium transfer, and culturing for 4d in an environment with the stirring speed of 190r/min and the temperature of 40 ℃ to obtain the thiobacillus acidophilus culture liquid.
(2) Preparation of composite microbial inoculum
Respectively adding the bacillus subtilis culture solution and the Acidithiobacillus acidophilus culture solution into a fermentation tank D according to the adding proportion of 75% and 25%, adding a carbonaceous filler (with the diameter of 5-10 mm and the specific surface area of about 280m 2/g) which is subjected to high-pressure steam sterilization (under the pressure of 0.1MPa, namely 103.4kPa (1.05 kg/cm 2) steam pressure, the temperature in a pot reaches 121 ℃ and is maintained for 15-30 min) according to the concentration of 50g/L, and culturing for 1-2 days in the environment with the stirring speed of 190r/min and the temperature of 35 ℃ until the bacterial liquid concentration in the culture solution reaches 1X 10 7 cfu/mL to obtain the first bacterial agent.
Adding a carbonaceous filler (with the diameter of 5-10 mm and the specific surface area of about 280m 2/g) which is subjected to high-pressure steam sterilization (under the pressure of 0.1MPa, namely 103.4kPa (1.05 kg/cm 2) steam pressure, the temperature in the pot reaches 121 ℃ and is maintained for 15-30 min) into a mycobacterium culture solution in a fermentation tank A according to the concentration of 50g/L, and culturing for 0.5-1 day in an environment with the stirring speed of 190r/min and the temperature of 30 ℃ until the concentration of bacterial liquid in the culture solution reaches 1X 10 5 cfu/mL to obtain a second bacterial agent.
Fourth, experiments on deodorization performance of biological deodorization composite microbial inoculant
Inoculating the first microbial inoculum into a biological filter (L2 m)B2mH3.3m, air volume 2000m 3/h, filler height 1.6m, residence time t=12s) lower half layer of filler, the inoculation amount is the first microbial inoculum containing carbon filler 2L per ten thousand cubic meters of odor, the specific method is as follows: the carbonaceous filler in the first microbial inoculum is uniformly dispersed in the lower half layer of the biological filter, and the bacterial liquid is uniformly sprayed on the top of the filler. After the inoculation of the first microbial inoculum is completed, the biological filter is started to run for 1-2 days, and then the second microbial inoculum is inoculated.
Inoculating a second microbial inoculum to the upper half layer of the biological filter filler, wherein the inoculation amount of the second microbial inoculum is 5L of carbon-containing filler with odor per ten thousand cubic meters, and the specific method comprises the following steps: the carbonaceous filler in the second microbial inoculum is uniformly dispersed in the upper half layer of the biological filter, and the bacterial liquid is uniformly sprayed on the top of the filler. After the inoculation of the second microbial inoculum is finished, the biological filter is started to run for 3-4 days, and the outlet can reach a stable state.
The removal of hydrogen sulfide using a blank column as a control is shown in figure 4. As shown in fig. 4, the biological deodorization composite bacterial agent can complete the domestication and the starting of the biological filter for only 4-6 days, 100% removal of hydrogen sulfide is realized, and the same effect can be achieved only by 12-14 days for natural domestication.
As can be seen from fig. 4, when the concentration of hydrogen sulfide suddenly increases from below 20ppm to 80-100ppm, the natural domesticated biological tower (i.e., blank tower) is greatly affected by concentration fluctuation, which is shown as exceeding the standard of the outlet hydrogen sulfide; the biological deodorization composite bacterial agent (experimental tower) is basically not influenced by the fluctuation of the concentration of the inlet air hydrogen sulfide, and can still maintain the removal rate of nearly 100 percent. The biological deodorization tower adopting the biological deodorization composite bacterial agent has the capability of resisting high-concentration air inlet fluctuation, strong impact load resistance and stable deodorization performance.
When the inlet air concentration is 80-100ppm, the concentration of hydrogen sulfide at the outlet of the natural domesticated biological deodorizing tower is 20-40ppm, the deodorizing efficiency is 60-80%, while the experimental tower adopting the biological deodorizing microbial inoculum can still maintain 100% of the deodorizing efficiency, and compared with the natural domesticated and running stable biological filter, the deodorizing efficiency is improved by 20% -40%
(V) experiment on abnormal condition resistance
(1) Resistance to severe cold
The composite microbial inoculum is applied to a biological filter of a sewage treatment plant in winter, data of a certain freezing weather after the composite microbial inoculum runs stably are intercepted and drawn as shown in a figure 5, and the day 11-25 is a subzero cold weather, and as can be seen from the figure 5, the deodorization efficiency of an experimental tower adopting the biological deodorization microbial inoculum can still maintain the removal efficiency of more than 99 percent, and the deodorization efficiency of the biological filter (a natural domestication tower) without inoculating the composite microbial inoculum is greatly influenced by weather.
(2) Resistance to prolonged idle of biological towers
And after the biological filter which is inoculated with the biological deodorization composite bacterial agent and stably operates for 2 months, air and water supply is stopped for 1 month, the biological filter is restarted, and the biological filter can be restored to a stable state only by 2 days.
Example 2
Mycobacterium, bacillus subtilis, and Thiobacillus thiooxidans were purchased from Beijing Bai Ou Bo Wei Biotechnology Co., ltd, numbered bio-101896, bio-52790, and bio-67436, respectively.
The enrichment culture method of each strain was the same as "(1) preparation of single strain" in the third section of example 1.
The preparation process of the composite microbial inoculum comprises the following steps:
Respectively adding 80% and 20% of bacillus subtilis culture solution and thiobacillus thiooxidans culture solution into a fermentation tank according to the adding proportion, adding a carbonaceous filler (with the diameter of 5-10 mm and the specific surface area of about 280m 2/g) subjected to high-pressure steam sterilization (under the pressure of 0.1MPa, namely 103.4kPa (1.05 kg/cm 2) steam pressure, the temperature in a pot reaching 121 ℃ and the maintenance of 15-30 min) according to the concentration of 50g/L, and culturing for 1-2 days in the environment with the stirring speed of 190r/min and the temperature of 35 ℃ until the bacterial liquid concentration in the culture solution reaches 10 7 cfu/mL to obtain the first bacterial agent.
In another fermentation tank, adding a carbonaceous filler (with the diameter of 5-10 mm and the specific surface area of about 280m 2/g) which is subjected to high-pressure steam sterilization (under the pressure of 0.1MPa, namely 103.4kPa (1.05 kg/cm 2) steam pressure, the temperature in the tank reaches 121 ℃ and is maintained for 15-30 min) into the mycobacterium culture solution according to the concentration of 50g/L, and culturing for 0.5-1 day in the environment with the stirring speed of 190r/min and the temperature of 30 ℃ until the concentration of the bacterium solution in the culture solution reaches 10 5 cfu/mL to obtain a second bacterium agent.
The performance of the biological deodorization composite microbial inoculum obtained in the embodiment is researched according to the method in the embodiment 1, and the result shows that in the embodiment, the biological deodorization tower inoculated with the microbial inoculum needs 8-9 days to finish domestication and starting of a biological filter, and the biological deodorization efficiency is improved by 20-30%; compared with a natural domesticated biological deodorization tower, the biological deodorization tower inoculated with the microbial inoculum can improve the stability of the deodorization efficiency in northern severe cold regions, the deodorization efficiency of the natural domesticated biological deodorization tower in extremely cold weather is reduced to 20-60% from 100% in a stable state, and the deodorization efficiency of the biological deodorization tower inoculated with the microbial inoculum is 70-85%, and is improved by 20-60% compared with the stability of the natural domesticated biological deodorization tower.
Comparative example
Mycobacterium, alicyclic acid Bacillus and acidophilic sulfide are purchased from Beijing Bai Ou Bo Wei Biotechnology Co., ltd, numbered as bio-101896, bio-56652 and bio-67436, respectively.
The same culture medium as in example 1 and example 2 was used for Mycobacterium and Acidophilic sulfide.
Alicyclic acid bacillus was cultured in YSG medium (inc. Of microorganism technology, inc. Of guangdong.
The enrichment culture method of each strain and the preparation method of the composite microbial inoculum are basically the same as those of example 1, except that the bacillus subtilis and the corresponding culture medium are replaced with the alicyclic acid bacillus and YSG culture medium.
The deodorizing performance and the abnormal condition resistance performance of the composite microbial inoculum of this comparative example were measured by the method of example 1, and the results are as follows:
The biological deodorization tower needs 10-12 days to finish the domestication of the deodorization microorganisms and the starting of the deodorization tower, and the deodorization efficiency of the biological deodorization tower cannot be maintained to be 100 percent stable when the biological deodorization tower is impacted against high-concentration hydrogen sulfide (from 10-20ppm to 70-100 ppm), and the deodorization efficiency is only 60-80 percent, which is reduced by 20-40 percent compared with the stable state of 100 percent. Compared with a natural domesticated biological deodorization tower, the biological deodorization tower inoculated with the microbial inoculum can improve the stability of the deodorization efficiency in northern severe cold regions, the deodorization efficiency of the natural domesticated biological deodorization tower in extremely cold weather is reduced to 20-60% from 100% in a stable state, the deodorization efficiency of the biological deodorization tower inoculated with the microbial inoculum is 60-70%, and the efficiency stability of the biological deodorization tower inoculated with the microbial inoculum is only improved by 10-30% compared with that of the biological deodorization tower naturally domesticated.
The foregoing examples are illustrative of the present invention and are not intended to be limiting, and any other substitutions, modifications, combinations, alterations, simplifications, etc. which do not depart from the spirit and principles of the present invention are intended to be within the scope of the present invention.
Claims (16)
1. The biological deodorization composite microbial inoculum is characterized by comprising a first microbial inoculum and a second microbial inoculum according to the volume ratio of (1-5): (3-10);
Wherein the first microbial inoculum is bacillus subtilis (Bacillus Subtilis) and thiobacillus thiooxidans (Acidithiobacillus Thiooxidans), and the concentration of bacterial liquid is 1 multiplied by 10 6~1×109 CFU/mL; the second microbial inoculum is Mycobacterium (Mycobacterium Sp.), and the concentration of the bacterial liquid is 1 multiplied by 10 4 ~1×106 CFU/mL;
Wherein, the mycobacterium, the bacillus subtilis and the thiobacillus thiooxidans are purchased from Beijing Bai Oubo Wei biotechnology Co., ltd, and the serial numbers are bio-101896, bio-52790 and bio-67436 respectively.
2. The biological deodorization composite microbial agent of claim 1, wherein the volume ratio of the first microbial agent to the second microbial agent is (1-2): (5-10).
3. The biological deodorization composite bacterial agent according to claim 1, wherein the volume ratio of the first bacterial agent to the second bacterial agent is 2:5.
4. The biological deodorization composite bacterial agent according to claim 1, wherein the bacterial liquid concentration of the first bacterial agent is 1 x 10 6~1×108 CFU/mL.
5. The biological deodorization composite bacterial agent according to claim 1, wherein the bacterial liquid concentration of the first bacterial agent is 1 x 10 7 CFU/mL.
6. The biological deodorization composite bacterial agent according to claim 1, wherein the bacterial liquid concentration of the second bacterial agent is 5 x 10 4 ~6×105 CFU/mL.
7. The biological deodorization composite bacterial agent according to claim 1, wherein the bacterial liquid concentration of the second bacterial agent is 1 x 10 5 CFU/mL.
8. The biological deodorization composite bacterial agent according to claim 1, wherein in the first bacterial agent, the colony ratio of bacillus subtilis and thiobacillus thiooxidans is (7-8): (1-5).
9. The biological deodorization composite bacterial agent according to claim 1, wherein in the first bacterial agent, the colony ratio of bacillus subtilis and thiobacillus thiooxidans is (7-8): (2-3).
10. The preparation method of the biological deodorization composite bacterial agent according to any one of claims 1 to 9, which is characterized by comprising the following steps:
(1) Respectively carrying out fermentation culture to obtain a mycobacterium culture solution, a bacillus subtilis culture solution and a thiobacillus thiooxidans culture solution;
(2) Mixing bacillus subtilis culture solution and thiobacillus thiooxidans culture solution, adding sterilized carbonaceous filler into the obtained mixed bacterial solution, and culturing to obtain a first microbial inoculum;
(3) And adding the sterilized carbonaceous filler into a mycobacterium culture solution, and culturing to obtain a second microbial inoculum.
11. The preparation method of claim 10, wherein in the step (2), the volume percentages of the bacillus subtilis culture solution and the thiobacillus thiooxidans culture solution are respectively 70-80% and 20-30%, the sum of the bacillus subtilis culture solution and the thiobacillus thiooxidans culture solution is 100%, and the adding proportion of the carbonaceous filler is 50-100 g of the carbonaceous filler per liter of the mixed bacterial solution; in the step (3), the adding proportion of the carbonaceous filler is 50-100 g of the carbonaceous filler per liter of the mycobacterium culture solution.
12. The preparation method of claim 10, wherein the carbonaceous filler has a diameter of 5-10 mm, a specific surface area of not less than 250m 2/g, and a hydrophilic surface.
13. The use method of the biological deodorization composite bacterial agent as claimed in any one of claims 1 to 9, which is characterized by comprising the following steps in sequence:
Step one, inoculating a first microbial inoculum to the lower half layer of the biological filter, which is close to the air inlet side, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor;
And step two, inoculating a second microbial inoculum to the upper half layer of the biological filter far away from the air inlet side, wherein the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor.
14. The method for using the biological deodorization composite bacterial agent as claimed in claim 13, which is characterized by comprising the following steps in sequence:
dispersing a carbon filler in a first microbial inoculum on the lower half layer of the biological filter near the air inlet side, spraying bacterial liquid in the first microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 1-2 days;
Dispersing the carbon filler in the second microbial inoculum on the upper half layer of the biological filter far away from the air inlet side, spraying bacterial liquid in the second microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 3 to 4 days.
15. The use method of the biological deodorization composite bacterial agent as claimed in any one of claims 1 to 9, which is characterized by comprising the following steps in sequence:
Step one, inoculating a first microbial inoculum to the upper half layer of the biological filter, which is close to the air inlet side, wherein the inoculation amount is 3-10L of the first microbial inoculum per ten thousand cubic meters of odor;
And step two, inoculating a second microbial inoculum to the lower half layer of the biological filter far away from the air inlet side, wherein the inoculation amount is 3-10L of the second microbial inoculum per ten thousand cubic meters of odor.
16. The method for using the biological deodorization composite bacterial agent according to claim 15, which is characterized by comprising the following steps in sequence:
Dispersing a carbon filler in a first microbial inoculum on the upper half layer of the biological filter near the air inlet side, spraying bacterial liquid in the first microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 1-2 days;
dispersing the carbon filler in the second microbial inoculum on the lower half layer of the biological filter far away from the air inlet side, spraying bacterial liquid in the second microbial inoculum on the surface of the carbon filler, and starting the biological filter to operate for 3 to 4 days.
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