CN114717152A - Composite functional microbial inoculum with pseudomonas stutzeri as carrier for in-situ remediation of contaminated soil and application thereof - Google Patents
Composite functional microbial inoculum with pseudomonas stutzeri as carrier for in-situ remediation of contaminated soil and application thereof Download PDFInfo
- Publication number
- CN114717152A CN114717152A CN202210434957.1A CN202210434957A CN114717152A CN 114717152 A CN114717152 A CN 114717152A CN 202210434957 A CN202210434957 A CN 202210434957A CN 114717152 A CN114717152 A CN 114717152A
- Authority
- CN
- China
- Prior art keywords
- microbial inoculum
- vts
- pseudomonas
- pseudomonas aeruginosa
- soil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002689 soil Substances 0.000 title claims abstract description 67
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 66
- 241000589614 Pseudomonas stutzeri Species 0.000 title claims abstract description 50
- 239000002131 composite material Substances 0.000 title claims abstract description 25
- 238000005067 remediation Methods 0.000 title claims abstract description 22
- 238000011065 in-situ storage Methods 0.000 title claims abstract description 17
- UDCXNLDCQOXJNC-UHFFFAOYSA-N 2-n,4-n,1,3-tetraphenyl-1,3,2,4-diazadiphosphetidine-2,4-diamine Chemical compound C=1C=CC=CC=1NP(N1C=2C=CC=CC=2)N(C=2C=CC=CC=2)P1NC1=CC=CC=C1 UDCXNLDCQOXJNC-UHFFFAOYSA-N 0.000 claims abstract description 45
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims abstract description 45
- 238000004321 preservation Methods 0.000 claims abstract description 13
- 238000000855 fermentation Methods 0.000 claims description 30
- 230000004151 fermentation Effects 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 25
- 238000001694 spray drying Methods 0.000 claims description 21
- 230000001580 bacterial effect Effects 0.000 claims description 18
- 239000007788 liquid Substances 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 12
- 239000003225 biodiesel Substances 0.000 claims description 9
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 8
- 239000000843 powder Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 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 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 6
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 235000005822 corn Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 5
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 4
- 235000010344 sodium nitrate Nutrition 0.000 claims description 4
- 239000004317 sodium nitrate Substances 0.000 claims description 4
- 241001052560 Thallis Species 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims description 2
- 241000209149 Zea Species 0.000 claims 2
- 241000894006 Bacteria Species 0.000 abstract description 28
- 229930195733 hydrocarbon Natural products 0.000 abstract description 21
- 150000002430 hydrocarbons Chemical class 0.000 abstract description 21
- 239000004215 Carbon black (E152) Substances 0.000 abstract description 14
- 230000015556 catabolic process Effects 0.000 abstract description 11
- 238000006731 degradation reaction Methods 0.000 abstract description 11
- 244000005700 microbiome Species 0.000 abstract description 6
- 230000004083 survival effect Effects 0.000 abstract description 3
- 230000002688 persistence Effects 0.000 abstract description 2
- FCBUKWWQSZQDDI-UHFFFAOYSA-N rhamnolipid Chemical compound CCCCCCCC(CC(O)=O)OC(=O)CC(CCCCCCC)OC1OC(C)C(O)C(O)C1OC1C(O)C(O)C(O)C(C)O1 FCBUKWWQSZQDDI-UHFFFAOYSA-N 0.000 description 10
- 239000000243 solution Substances 0.000 description 10
- 239000003209 petroleum derivative Substances 0.000 description 9
- 239000000306 component Substances 0.000 description 8
- 239000004094 surface-active agent Substances 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 230000007613 environmental effect Effects 0.000 description 5
- 239000007789 gas Substances 0.000 description 5
- 230000000813 microbial effect Effects 0.000 description 5
- 241000194107 Bacillus megaterium Species 0.000 description 4
- 240000008042 Zea mays Species 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 230000000593 degrading effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000000644 propagated effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000969 carrier Substances 0.000 description 3
- 238000009792 diffusion process Methods 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 239000008394 flocculating agent Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000011020 pilot scale process Methods 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 239000005913 Maltodextrin Substances 0.000 description 1
- 229920002774 Maltodextrin Polymers 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000004115 Sodium Silicate Substances 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000589651 Zoogloea Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 230000001094 effect on targets Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000000855 fungicidal effect Effects 0.000 description 1
- 239000000417 fungicide Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 229940035034 maltodextrin Drugs 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 238000013048 microbiological method Methods 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
- 229910052911 sodium silicate Inorganic materials 0.000 description 1
- 244000000000 soil microbiome Species 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Images
Classifications
-
- 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
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
-
- 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
Landscapes
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Soil Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Processing Of Solid Wastes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention discloses a composite functional microbial inoculum for in-situ remediation of contaminated soil by taking pseudomonas stutzeri as a carrier and application thereof, wherein the composite functional microbial inoculum comprises pseudomonas stutzeri VTCQ-1 and pseudomonas aeruginosa VTS-1, the preservation number of the pseudomonas stutzeri VTCQ-1 is CGMCC24469, the preservation number of the pseudomonas aeruginosa VTS-1 is CGMCC2200, and the pseudomonas stutzeri VTCQ-1 is used as the carrier of the pseudomonas aeruginosa VTS-1. The invention utilizes the characteristics that the specific surface area of the colony of the pseudomonas stutzeri VTCQ-1 is larger and a larger and stable attachment place can be provided for other bacteria, the pseudomonas stutzeri VTS-1 is mixed and cultured with the pseudomonas aeruginosa VTS-1, the survival rate of the pseudomonas aeruginosa VTS-1 can be improved, the prepared composite functional microbial inoculum can improve the degradation efficiency of hydrocarbon in soil, greatly shorten the treatment period of the hydrocarbon polluted soil, and maintain the diversity and the persistence of soil microorganisms.
Description
Technical Field
The invention relates to the field of bioremediation of petroleum hydrocarbon contaminated soil, in particular to a composite functional microbial inoculum for in-situ remediation of contaminated soil by taking pseudomonas stutzeri as a carrier and application thereof.
Background
At present, methods such as physical, chemical and microbiological methods are more in the field of in-situ remediation of hydrocarbon-contaminated soil, wherein the microbial in-situ remediation of the hydrocarbon-contaminated soil is the most environmentally-friendly, economical and reliable treatment mode without secondary pollution and with sustainability and environmental friendliness, and the method attracts wide attention and intensive research in various environmental protection fields in the world.
The key of the microbial remediation method is the screening of efficient hydrocarbon degrading bacteria, the maintenance activity of the flora in the soil environment, the body density of the microbial inoculum, the diffusion and propagation of the microbial inoculum in the soil environment, and the capture and aggregation of hydrocarbon pollutants. In order to meet the requirements, the method of increasing the input amount of the microbial inoculum, improving the injection of nutrient components, adding a flocculating agent or a surfactant and the like is generally adopted to assist in improving the hydrocarbon treatment efficiency. The cost is increased by increasing the input amount of the microbial inoculum, and chemical components such as flocculating agents, surfactants and the like added into the microbial inoculum are especially undegraded parts or cause secondary environmental pollution. In addition, the composite microbial inoculum carrier generally uses one or more of calcium carbonate, diatomite, sodium silicate, maltodextrin, sucrose, glucose, peptone and the like which are mixed according to a proportion, but the carriers have the quantitative characteristic, once the microbial inoculum is dissolved or microorganisms enter the soil environment and are completely released, the microorganisms can not be diffused by the aid of the carriers, and the carriers have limited addition amount and can not assist the microorganisms to complete the aggregation and degradation of hydrocarbons in a long time and a large range.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a composite functional microbial inoculum for in-situ remediation of polluted soil by taking pseudomonas stutzeri as a carrier and application thereof, so as to solve the problem that the carrier of the composite microbial inoculum cannot assist microorganisms to complete accumulation and degradation of hydrocarbons in a large range for a long time, and further improve the degradation efficiency of petroleum hydrocarbons in soil.
The invention is realized by the following steps:
in a first aspect, the invention provides a composite functional microbial inoculum, which comprises pseudomonas stutzeri VTCQ-1 and pseudomonas aeruginosa VTS-1;
the preservation number of the Pseudomonas stutzeri VTCQ-1 is CGMCC 24469;
the preservation number of the pseudomonas aeruginosa VTS-1 is CGMCC 2200;
pseudomonas stutzeri VTCQ-1 is used as a vector of Pseudomonas aeruginosa VTS-1.
In a second aspect, the invention provides a preparation method of a composite functional microbial inoculum, which comprises the step of carrying out continuous mixed culture on Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1.
In a third aspect, the invention provides a method for in-situ remediation of contaminated soil, which comprises the step of remedying the soil to be remediated by using the complex functional microbial inoculum.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a composite functional microbial inoculum for in-situ remediation of contaminated soil by taking pseudomonas stutzeri as a carrier, which utilizes the characteristics that the specific surface area of a pseudomonas stutzeri VTCQ-1 colony is larger and a larger and stable attachment place can be provided for other bacteria, and the microbial inoculum is mixed and cultured with pseudomonas aeruginosa VTS-1, so that the pseudomonas stutzeri VTCQ-1 colony can wrap the pseudomonas aeruginosa VTS-1, and the survival rate of the VTS-1 is improved in the process of preparing the microbial inoculum by a spray drying method. The pseudomonas stutzeri VTCQ-1 is screened and domesticated from a soil environment, has good environmental adaptability, can be rapidly propagated in an underground deep soil environment, has a large diffusion range in the soil environment, provides more attachment sites for other composite functional microbial agents which are simultaneously propagated, and is easy to form relatively large-range and relatively large-quantity microbial colonies; meanwhile, the pseudomonas stutzeri VTCQ-1 can also be adsorbed on the surface of hydrocarbon-polluted soil to fix functional bacteria, adsorb hydrocarbons on the surface of soil particles, play a role in gathering hydrocarbon substances for other complex functional bacteria with higher degradation efficiency in the soil, can be compared with a mobile restaurant in which soil remediation functional bacteria degrade hydrocarbons in the soil, and can infinitely expand the operating range of the mobile restaurant. The composite functional microbial inoculum provided by the invention can improve the degradation efficiency of hydrocarbon in soil, greatly shorten the treatment period of hydrocarbon-polluted soil and maintain the diversity and the persistence of soil microorganisms.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts. FIG. 1 shows the detection results of rhamnolipid surfactant component in fermentation broth according to example 1 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
Some embodiments of the invention provide a composite functional microbial inoculum, which comprises Pseudomonas stutzeri VTCQ-1(Pseudomonas stutzeri) and Pseudomonas aeruginosa VTS-1(Pseudomonas aeruginosa), wherein the preservation number of the Pseudomonas stutzeri VTCQ-1 is CGMCC 24469; the preservation number of the pseudomonas aeruginosa VTS-1 is CGMCC 2200; pseudomonas stutzeri VTCQ-1 is used as a vector of Pseudomonas aeruginosa VTS-1.
The preservation information of the Pseudomonas stutzeri VTCQ-1 is as follows: the preservation number is CGMCC 24469; classified and named as Pseudomonas stutzeri; the preservation unit is China general microbiological culture Collection center; the preservation address is the microbiological research institute of China academy of sciences No. 3, Xilu No.1, Beijing, facing Yang district; the preservation time is 03/07/2022.
The Pseudomonas aeruginosa VTS-1 is disclosed in the patent document CN101177696B and is an existing deposited strain.
The Pseudomonas stutzeri VTCQ-1 is obtained by multiple times of repeated acclimation and separation from a water layer of deep contaminated soil of a certain gas station, and the morphological characteristics of the Pseudomonas stutzeri VTCQ-1 are as follows: the colony is round, the edge is irregular, the surface is wrinkled, the specific surface area is large, the colony is white waxy, the colony is easy to be attached to the surface of a common LB solid medium or the surface of other solid matters, and the colony is easy to be peeled off in a sheet; observing the shape of the thallus under a microscope oil microscope, wherein the thallus is in a short rod shape with the length of 0.7-3.6 mu m and the width of 0.35-42 mu m; gram-negative bacteria. 16sRNA sequencing is carried out on the pure strain obtained after screening and separation, and the 16sRNA gene sequence of the strain is shown in a sequence table SEQ ID No. 1:
AGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGG ATGAGTGGAGCTTGCTCCATGATTCAGCGGCGGACGGGTGAGTAATG CCTAGGAATCTGCCTGGTAGTGGGGGACAACGTTTCGAAAGGAACGC TAATACCGCATACGTCCTACGGGAGAAAGCGGGGGACCTTCGGACCT CACGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAAA GGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGT CACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAG TGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCG TGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGA AGGGCAGTAAGTTAATACCTTGCTGTTTTGACGTTACCAACAGAATAA GCACCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCA AGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTCGTT AAGTTGGATGTGAAAGCCCCGGGCTCAACCTGGGAACTGCATCCAAA ACTGGCGAGCTAGAGTATGGCAGAGGGTGGTGGAATTTCCTGTGTAG CGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGAC CACCTGGGCTAATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAA CAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGC CGTTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCATTAAGTCGA CCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACG GGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGC GAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGG ATTGGTGCCTTCGGGAACTCTGACACAGGTGCTGCATGGCTGTCGTCA GCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCC TTGTCCTTAGTTACCAGCACGTTAAGGTGGGCACTCTAAGGAGACTGC CGGTGACAAACCGGAGGAAGGGTGGGGATGACGTCAAGTCATCATGG CCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGG GTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTC CGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTA ATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACAC ACCGCCCGTCACACC。
the strain is determined to belong to Pseudomonas stutzeri by comparison and is named as Pseudomonas stutzeri VTCQ-1.
And the morphological characteristics of the pseudomonas aeruginosa VTS-1 are as follows: the bacterial colony is round and white, and has a non-glossy surface and a neat edge. The microscopic examination shows that the short bacillus has a length of 0.66-1.31 μm and a width of 0.4-0.51 μm, and gram-negative bacteria.
The inventor of the invention researches and discovers that surface-wrinkled colonies can easily grow out of Pseudomonas stutzeri VTCQ-1 in a liquid and solid culture medium, and the morphological characteristics of the colonies cause that the colonies have the characteristic of larger specific surface area and can provide larger and stable attachment sites for other bacteria; after the pseudomonas stutzeri VTCQ-1 bacterial colony and the pseudomonas aeruginosa VTS-1 are mixed, the pseudomonas stutzeri VTCQ-1 bacterial colony wraps the pseudomonas aeruginosa VTS-1, so that the survival rate of the VTS-1 can be improved in the process of preparing the microbial inoculum by a spray drying method; the pseudomonas stutzeri is obtained by screening and domesticating soil environment, has good environmental adaptability, can be rapidly propagated in underground deep soil environment, has a large diffusion range in the soil environment, provides more attachment sites for other complex functional bacteria simultaneously propagated, and is easy to form relatively large-range and relatively large-quantity zoogles; the adhesion of the pseudomonas stutzeri VTCQ-1 colony can adsorb functional bacteria to form a zoogloea, can adsorb functional bacteria fixed on the surface of hydrocarbon-polluted soil, adsorbs hydrocarbons on the surface of soil particles, plays a role in gathering hydrocarbon substances for other composite functional bacteria agents with higher degradation efficiency in the soil, can be compared with a flowing restaurant for degrading the hydrocarbons in the soil by using soil remediation functional bacteria agents, and can infinitely expand the operating range of the flowing restaurant.
Preferably, the density of the VTS-1 bacteria of the pseudomonas aeruginosa in the composite microbial inoculum is 8.0 multiplied by 1011- 9.45×1011Per gram.
In addition, the VTS-1 fermentation process can generate a surfactant, can reduce the surface tension of the liquid, is beneficial to the release of petroleum hydrocarbon on the surface of soil particles and promotes the degradation of bacteria to the hydrocarbon. In some embodiments, the VTS-1 fermentation process results in a rhamnolipid surfactant component comprising primarily rhamnolipids of the monosaccharide C10, C12 and disaccharide C10, C12 structures.
Some embodiments of the invention provide a preparation method of a composite functional microbial inoculum, which comprises the step of carrying out continuous mixed culture on Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1.
Preferably, the culture medium for the continuous mixed culture comprises: 30-50g/L glucose, 12-17g/L sodium nitrate, 6-7g/L potassium dihydrogen phosphate, 4-5g/L disodium hydrogen phosphate and 1.5-2.5g/L peptone.
Preferably, the pH of the medium is 7.0-7.2.
Preferably, the culture medium is also added with 15-20ml/L biodiesel.
According to the invention, the biodiesel is used as a part of the carbon source of the fermentation culture medium VTS-1 of the pseudomonas aeruginosa for the thallus fermentation culture, so that the cost is saved, the degradation of the hydrocarbon degrading bacteria to petroleum hydrocarbon can be adapted in the fermentation stage, and the degradation effect on target pollutants in the soil can be more remarkable after the biodiesel is applied to the hydrocarbon-polluted soil.
Preferably, the continuous mixed culture comprises two stages, wherein the first stage is to inoculate pseudomonas aeruginosa for culture, and the second stage is to inoculate pseudomonas stutzeri for mixed culture after the first stage of culture.
Preferably, the inoculation amount of the Pseudomonas aeruginosa VTS-1 is 2-5%.
Preferably, the inoculum size of Pseudomonas stutzeri VTCQ-1 is 2% -5%.
Preferably, the culture temperature of the first stage is: the constant temperature is 37 ℃, the stirring speed is 150-3Min, the tank pressure is 0.03-0.05MPa, and the fermentation is continued for 51-56 h.
Preferably, the culture temperature in the second stage is: the constant temperature is 35 ℃, the stirring speed is 180-200rpm, and the air quantity is 0.6-0.7m3Min, the tank pressure is 0.03-0.05MPa, and the fermentation is continued for 22-24 h.
In the process of culturing the two bacteria, the oil gas removing filter is arranged at the exhaust port of the fermentation tank, so that the volatile biodiesel before degradation can be recovered, and the atmospheric pollution caused by volatile gas in the biodiesel can be avoided.
Preferably, the preparation method further comprises adding the corn steep liquor dry powder into the mixed bacterial liquid and then performing spray drying.
Preferably, the addition amount of the corn steep liquor dry powder is 170-190 g/L.
Preferably, the conditions for spray drying are an inlet temperature of 110-120 ℃ and an outlet temperature of 50-60 ℃.
The invention also provides a method for in-situ remediation of contaminated soil, which comprises the step of remedying the soil to be remediated by adopting the composite functional microbial inoculum.
When the complex functional microbial inoculum is used for restoration, the complex functional microbial inoculum is added into water according to the addition concentration of 0.5-0.8 per mill wt to prepare a microbial inoculum use solution, and then the microbial inoculum use solution is used for soil restoration.
Preferably, the using amount of the microbial inoculum using solution is 25-30% of the volume of the polluted soil.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The embodiment provides a preparation method of a complex functional microbial inoculum, which comprises the following steps:
s1, preparing a fermentation culture medium
40g of glucose, 15g of sodium nitrate, 6.5g of potassium dihydrogen phosphate, 6g of disodium hydrogen phosphate, 2g of peptone and 340ml of biodiesel are respectively added into a 30L pilot-scale fermentation tank, and water is added to the tank until the volume is 17L.
In this example, a deoiling filter was installed at the exhaust port of the fermentation tank, and after the fermentation tank was sterilized conventionally, the pH of the culture medium was adjusted to 7.0 with 20% by mass sodium hydroxide.
S2, culturing Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1
Firstly, 425ml of pseudomonas aeruginosa strain VTS-1 is inoculated, and the culture conditions are as follows: keeping the temperature at 37 ℃, stirring at 190rpm, and blowingAmount of 0.3m3Min, the tank pressure is 0.04MPa, and the fermentation is continued for 54 h;
after 54h of fermentation, 425ml of Pseudomonas stutzeri VTCQ-1 is inoculated into the fermentation tank, and the culture conditions are as follows: keeping the temperature at 35 ℃, stirring speed at 180rpm and ventilation quantity at 0.65m3And/min, the tank pressure is 0.03MPa, and the fermentation is finished after the fermentation culture is continued for 23 hours to obtain mixed bacteria liquid.
And (3) detecting the mixed bacterial liquid by using a surface tension meter when the mixed bacterial liquid is placed in a tank, reducing the surface tension of the bacterial liquid, containing surface active substances, centrifugally concentrating the bacterial liquid, and detecting by using a liquid-mass spectrometer to find that the fermentation liquid contains rhamnolipid surfactant components which mainly comprise R1, R3 and R4 type rhamnolipid. As a result of detection, the relative proportion among the rhamnolipid configurations in the sample is about R1 type: r3 type: the R4 type is 27:31:15, the proportion of the R1 type rhamnolipid in the sample is calculated to be 4.2-4.6%, the proportion of the R3 type rhamnolipid in the sample is 4.8-5.2%, and the proportion of the R4 type rhamnolipid in the sample is 1.2-1.6%. Rhamnolipid accounts for 10.3-10.7% of the total amount of the sample. As shown in figure 1 below.
S3, spray drying to prepare the microbial inoculum
Adding 3.2kg of corn steep liquor dry powder into the mixed bacterial liquid obtained in the step S2, dissolving and mixing uniformly, and carrying out high-temperature spray drying under the spray drying conditions: the inlet temperature is 115 ℃, the outlet temperature is 55 ℃, the spray drying is finished to obtain a brown yellow powdery microbial inoculum product, and the density of the dilution plate counting pseudomonas aeruginosa strain VTS-1 is 9.45 multiplied by 1011About one per gram (mu/g).
Example 2
The embodiment provides a preparation method of a complex functional microbial inoculum, which comprises the following steps:
s1, preparing a fermentation culture medium
To 30m340kg of glucose, 15kg of sodium nitrate, 6.5kg of potassium dihydrogen phosphate, 6kg of disodium hydrogen phosphate, 2kg of peptone and 360L of biodiesel are respectively added into a pilot-scale fermentation tank, and water is added to the tank until the volume is 18m3。
In this example, a deoiling filter was installed at the exhaust port of the fermentation tank, and after the fermentation tank was sterilized conventionally, the pH of the culture medium was adjusted to 7.0 with 20% by mass sodium hydroxide.
S2, culturing Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1
First, access 0.4m3The culture conditions of the pseudomonas aeruginosa strain VTS-1 are as follows: keeping the temperature at 37 ℃, stirring at 200rpm and air volume at 0.3m3Min, the tank pressure is 0.04MPa, and the fermentation is continued for 54 h;
fermenting for 54h, and inoculating 0.4m3Pseudomonas stutzeri VTCQ-1, the culture conditions at this time were: keeping the temperature at 35 ℃, stirring speed at 180rpm and ventilation quantity at 0.65m3And/min, the tank pressure is 0.03MPa, and the fermentation is finished after the fermentation culture is continued for 23 hours to obtain mixed bacteria liquid.
S3, spray drying to prepare the microbial inoculum
Adding 3.42 tons of corn steep liquor dry powder into the mixed bacterial liquid obtained in the step S2, dissolving and mixing uniformly, and carrying out high-temperature spray drying under the spray drying conditions: the inlet temperature is 115 ℃, the outlet temperature is 55 ℃, the spray drying is finished to obtain a brown yellow powdery microbial inoculum product, and the microbial density of the dilution plate counting pseudomonas aeruginosa VTS-1 is 9.45 multiplied by 1011About one per gram (mu/g).
Example 3
The embodiment provides a method for in-situ remediation of contaminated soil, which comprises the following steps:
adding the microbial inoculum product obtained in the embodiment 1 into water according to the dosage of 0.5 per mill, and uniformly stirring to obtain a microbial inoculum use solution;
then taking back the polluted soil underground from a certain gas station (the average petroleum hydrocarbon content of a three-dimensional sampling point in a pollution range is 1328 mg/m)3) And putting the mixture into a glass column with the diameter of 0.25m and the height of 1.0m, and compacting the mixture to be close to the original existing state of the soil. A metal pipe with a plurality of holes on the pipe wall is punched from the middle and inserted into the center of soil in the device, and 40L of microbial inoculum solution is injected. And regularly detecting the thallus density and the petroleum hydrocarbon content of the multi-site soil sample.
After the microbial inoculum is added into soil, the two bacteria are dominant flora, and the density of the bacteria is 10 from the beginning5Per g is increased to 108The cell density is maintained at 6.8 × 10 within 1-12 months7-7.5×108Per gram, two kinds of bacteria are always used as dominant bacteria;the hydrocarbon treatment rate is 91.36% in 12 months, the treatment rate of the pseudomonas aeruginosa microbial inoculum used alone is 86.36%, and the treatment rate is improved by about 5% compared with the treatment rate of the hydrocarbon degrading bacteria used alone.
Example 4
The embodiment provides a method for in-situ remediation of contaminated soil, which comprises the following steps:
adding the microbial inoculum product obtained in the embodiment 2 into water according to the dosage of 0.4 per mill, and uniformly stirring to obtain a microbial inoculum use solution;
then the polluted soil (the average petroleum hydrocarbon content of a three-dimensional sampling point in a pollution range is 1418 mg/m) retrieved from the underground of a certain gas station3) Putting into a glass column with diameter of 0.25m and height of 1.0m, and compacting to approximate the original existing state of soil. And (3) punching a metal pipe with multiple holes on the pipe wall from the middle, inserting the metal pipe into the soil center in the device, injecting 27% of microbial inoculum use solution according to the calculated pollution range volume, and periodically detecting the thallus density and the petroleum hydrocarbon content of the soil sample at multiple sites.
The density of the thalli in the soil is 2.5 multiplied by 10 after the microbial inoculum is injected for 7 days5The amount per gram is increased to 8.9 multiplied by 108Cell/g, density of soil bacteria at 12 months 8.25X 107The hydrocarbon treatment rate per gram at 12 months is 92.3 percent.
Comparative example 1
The pseudomonas aeruginosa VTS-1 is independently inoculated into a culture medium for culture, and the obtained bacterium liquid is sprayed and dried to prepare the bactericide after being placed in a tank, wherein the components of the culture medium, the culture conditions and the spray drying strip are the same as those in the embodiment 1. The density of the pseudomonas aeruginosa VTS-1 in the prepared microbial inoculum is 9.10 multiplied by 1011About one per gram (mu/g).
Comparative example 2
Inoculating Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1 into culture medium for culturing, placing into a jar, spray drying the obtained bacterial liquid to obtain bacterial preparation, wherein the culture medium components and spray drying conditions are the same as those of example 1, the culture conditions are constant temperature of 37 deg.C, stirring at 200rpm, and air volume of 0.3m3Min, the tank pressure is 0.04MPa, and the fermentation is continued for 48 h. The thallus density of the Pseudomonas aeruginosa VTS-1 in the microbial inoculum prepared by the culture conditions is 7.24 multiplied by 1011About one/g, the content of the surface active substances in the bacterial liquid is lower.
Comparative example 3
The pseudomonas stutzeri VTCQ-1 is mixed with bacillus megaterium for culture, and the components of a 17L culture medium are as follows: 40g of glucose, 15g of ammonium chloride, 6.5g of potassium dihydrogen phosphate, 6g of dipotassium hydrogen phosphate, 2g of peptone and 2g of yeast extract powder, wherein the culture conditions are as follows: keeping the temperature at 35 ℃, stirring at 200rpm and air quantity at 0.4m3The culture time is 18-22h, the spray drying condition is the same as that of example 1, and the culture effect is that the density of the bacillus megaterium in the fungicide is 4.25 multiplied by 1011The cell/g is increased by 11 percent compared with that of the single-cultured bacillus megaterium, and the cell density of the single-cultured bacillus megaterium is 3.78 multiplied by 1011About one per gram.
Comparative example 4
The microbial inoculum prepared in the comparative example 1 is used for in-situ remediation of the polluted soil, and the remediation method is the same as that in the example 3. The hydrocarbon treatment rate of the microbial inoculum is 86.36 percent 12 months after the microbial inoculum is added into the soil.
In conclusion, the method realizes the multiplication of pseudomonas aeruginosa VTS-1 in the fermentation culture process by using biodiesel as a carbon source and can generate the rhamnolipid surfactant, VTCQ-1 is used as a carrier for spray drying of VTS-1 pseudomonas aeruginosa, the thallus density of the microbial inoculum is 3 percent higher than that of the single spray drying of the pseudomonas aeruginosa, and the total thallus density of the two strains after spray drying is 9.45 multiplied by 1011Per gram; the microbial inoculum is added into petroleum hydrocarbon soil, and the density of the soil thalli is 10 from the beginning5Per g is increased to 108The cell density is maintained at 6.8 × 10 within 1-12 months after 7 days per gram7-8.25×108Each gram of the strain is obtained by taking two kinds of bacteria as dominant bacteria all the time; the petroleum hydrocarbon treatment rate of 12 months is 91.36 percent, which is at least 5 percent higher than that of VTS-1 alone, and is higher than that of other microbial inoculum currently reported.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Sequence listing
<110> Beijing Wauteris environmental protection technology development Limited
<120> composite functional microbial inoculum for in-situ remediation of contaminated soil by taking pseudomonas stutzeri as carrier and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1383
<212> DNA
<213> Pseudomonas stutzeri
<400> 1
agattgaacg ctggcggcag gcctaacaca tgcaagtcga gcggatgagt ggagcttgct 60
ccatgattca gcggcggacg ggtgagtaat gcctaggaat ctgcctggta gtgggggaca 120
acgtttcgaa aggaacgcta ataccgcata cgtcctacgg gagaaagcgg gggaccttcg 180
gacctcacgc tatcagatga gcctaggtcg gattagctag ttggtgaggt aaaggctcac 240
caaggcgacg atccgtaact ggtctgagag gatgatcagt cacactggaa ctgagacacg 300
gtccagactc ctacgggagg cagcagtggg gaatattgga caatgggcga aagcctgatc 360
cagccatgcc gcgtgtgtga agaaggtctt cggattgtaa agcactttaa gttgggagga 420
agggcagtaa gttaatacct tgctgttttg acgttaccaa cagaataagc accggctaac 480
ttcgtgccag cagccgcggt aatacgaagg gtgcaagcgt taatcggaat tactgggcgt 540
aaagcgcgcg taggtggttc gttaagttgg atgtgaaagc cccgggctca acctgggaac 600
tgcatccaaa actggcgagc tagagtatgg cagagggtgg tggaatttcc tgtgtagcgg 660
tgaaatgcgt agatatagga aggaacacca gtggcgaagg cgaccacctg ggctaatact 720
gacactgagg tgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc 780
gtaaacgatg tcgactagcc gttgggatcc ttgagatctt agtggcgcag ctaacgcatt 840
aagtcgaccg cctggggagt acggccgcaa ggttaaaact caaatgaatt gacgggggcc 900
cgcacaagcg gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggcct 960
tgacatgcag agaactttcc agagatggat tggtgccttc gggaactctg acacaggtgc 1020
tgcatggctg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgta acgagcgcaa 1080
cccttgtcct tagttaccag cacgttaagg tgggcactct aaggagactg ccggtgacaa 1140
accggaggaa gggtggggat gacgtcaagt catcatggcc cttacggcct gggctacaca 1200
cgtgctacaa tggtcggtac aaagggttgc caagccgcga ggtggagcta atcccataaa 1260
accgatcgta gtccggatcg cagtctgcaa ctcgactgcg tgaagtcgga atcgctagta 1320
atcgtgaatc agaatgtcac ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac 1380
acc 1383
Claims (10)
1. A composite functional microbial inoculum is characterized by comprising Pseudomonas stutzeri VTCQ-1 and Pseudomonas aeruginosa VTS-1;
the preservation number of the Pseudomonas stutzeri VTCQ-1 is CGMCC 24469;
the preservation number of the pseudomonas aeruginosa VTS-1 is CGMCC 2200;
the Pseudomonas stutzeri VTCQ-1 is used as a vector of the Pseudomonas aeruginosa VTS-1.
2. The composite functional microbial inoculum according to claim 1, wherein the density of the pseudomonas aeruginosa VTS-1 thalli in the composite microbial inoculum is 8.0 x 1011-9.45×1011Per gram.
3. A method for preparing a composite functional microbial inoculum, which is characterized by comprising the step of carrying out continuous mixed culture on the Pseudomonas stutzeri VTCQ-1 and the Pseudomonas aeruginosa VTS-1 according to claim 1.
4. The method for preparing a complex functional bacterial agent according to claim 3, wherein the culture medium for continuous mixed culture comprises: 30-50g/L glucose, 12-17g/L sodium nitrate, 6-7g/L potassium dihydrogen phosphate, 4-5g/L disodium hydrogen phosphate and 1.5-2.5g/L peptone;
preferably, the pH of the culture medium is 7.0-7.2;
preferably, the culture medium is added with 15-20ml/L biodiesel.
5. The method for preparing the composite functional microbial inoculum according to claim 4, wherein the mixed bacterial liquid is obtained by the continuous mixed culture, the continuous mixed culture comprises two stages, the first stage is to firstly inoculate pseudomonas aeruginosa for culture, and the second stage is to inoculate pseudomonas stutzeri for mixed culture on the basis of the first stage culture;
preferably, the inoculation amount of the pseudomonas aeruginosa is 2% -5%;
preferably, the inoculum size of the pseudomonas stutzeri is 2% -5%.
6. The method for preparing a complex functional bacterial agent according to claim 5, wherein the culture temperature in the first stage is: the constant temperature is 37 ℃, the stirring speed is 150-3Min, the tank pressure is 0.03-0.05MPa, and the fermentation is continued for 51-56 h;
the culture temperature of the second stage is as follows: the constant temperature is 35 ℃, the stirring speed is 180-200rpm, and the air quantity is 0.6-0.7m3Min, the tank pressure is 0.03-0.05MPa, and the fermentation is continued for 22-24 h.
7. The method for preparing a complex functional bacterial agent according to claim 5, further comprising adding corn steep liquor dry powder to the mixed bacterial liquid and then performing spray drying;
preferably, the addition amount of the corn steep liquor dry powder is 170-190 g/L;
preferably, the conditions for spray drying are an inlet temperature of 110-120 ℃ and an outlet temperature of 50-60 ℃.
8. A method for in-situ remediation of contaminated soil, which comprises the step of remediating soil to be remediated with the complex functional microbial inoculum according to claim 1.
9. The method for in-situ remediation of contaminated soil according to claim 8, wherein the method comprises the steps of adding the complex microbial inoculum into water according to an addition concentration of 0.5 to 0.8 per thousand wt to prepare a microbial inoculum use solution, and then using the microbial inoculum use solution for soil remediation.
10. The method for in-situ remediation of contaminated soil according to claim 9, wherein the microbial inoculum use solution is used in an amount of 25-30% by volume of the contaminated soil.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210434957.1A CN114717152B (en) | 2022-04-24 | 2022-04-24 | Composite functional microbial agent taking pseudomonas stutzeri as carrier for in-situ remediation of polluted soil and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210434957.1A CN114717152B (en) | 2022-04-24 | 2022-04-24 | Composite functional microbial agent taking pseudomonas stutzeri as carrier for in-situ remediation of polluted soil and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114717152A true CN114717152A (en) | 2022-07-08 |
| CN114717152B CN114717152B (en) | 2024-01-09 |
Family
ID=82245217
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210434957.1A Active CN114717152B (en) | 2022-04-24 | 2022-04-24 | Composite functional microbial agent taking pseudomonas stutzeri as carrier for in-situ remediation of polluted soil and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114717152B (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2002122612A (en) * | 2002-08-23 | 2004-03-10 | Научно-исследовательский центр токсикологии и гигиенической регламентации биопрепаратов | The bacterial strain Pseudomonas stutzeri MEV-S1, used for the purification of soils, ground and surface water from oil and products of its processing |
| CN101177696A (en) * | 2007-11-05 | 2008-05-14 | 大庆沃太斯化工有限公司 | Industrial preparation method of rhamnolipid biological fermentation liquor |
| CN104276738A (en) * | 2014-09-29 | 2015-01-14 | 西安建筑科技大学 | Rapid harmless biological degradation method against waxy oil sludge |
| CN106635856A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Open culture method and application of pseudomonas stutzeri FSTB-5 |
| CN108102979A (en) * | 2018-02-06 | 2018-06-01 | 北京大学 | A kind of degradation bacteria strains JN5 of oily sludge petrochina hydro carbons and its application |
| CN108546659A (en) * | 2018-04-12 | 2018-09-18 | 陕西省微生物研究所 | A kind of alkane and degrading polycyclic aromatic hydrocarbons composite bacteria agent and preparation method thereof |
| EP3626683A1 (en) * | 2018-09-07 | 2020-03-25 | INDIAN OIL CORPORATION Ltd. | A process for bio-sludge reduction in hydrocarbon refinery effluent treatment plant through microbial interventions |
-
2022
- 2022-04-24 CN CN202210434957.1A patent/CN114717152B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2002122612A (en) * | 2002-08-23 | 2004-03-10 | Научно-исследовательский центр токсикологии и гигиенической регламентации биопрепаратов | The bacterial strain Pseudomonas stutzeri MEV-S1, used for the purification of soils, ground and surface water from oil and products of its processing |
| CN101177696A (en) * | 2007-11-05 | 2008-05-14 | 大庆沃太斯化工有限公司 | Industrial preparation method of rhamnolipid biological fermentation liquor |
| CN104276738A (en) * | 2014-09-29 | 2015-01-14 | 西安建筑科技大学 | Rapid harmless biological degradation method against waxy oil sludge |
| CN106635856A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Open culture method and application of pseudomonas stutzeri FSTB-5 |
| CN108102979A (en) * | 2018-02-06 | 2018-06-01 | 北京大学 | A kind of degradation bacteria strains JN5 of oily sludge petrochina hydro carbons and its application |
| CN108546659A (en) * | 2018-04-12 | 2018-09-18 | 陕西省微生物研究所 | A kind of alkane and degrading polycyclic aromatic hydrocarbons composite bacteria agent and preparation method thereof |
| EP3626683A1 (en) * | 2018-09-07 | 2020-03-25 | INDIAN OIL CORPORATION Ltd. | A process for bio-sludge reduction in hydrocarbon refinery effluent treatment plant through microbial interventions |
Non-Patent Citations (1)
| Title |
|---|
| V. P. JAYACHANDRAN ET AL.: "Degradation of 3-chlorobenzoate and phenol singly and in mixture by a mixed culture of two ortho-pathway-following Pseudomonas strains", 《J IND MICROBIOL BIOTECHNOL》, vol. 36, pages 219 - 227, XP019665551 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114717152B (en) | 2024-01-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104312938B (en) | Pseudomonas putida strain and fungicide and application of pseudomonas putida strain | |
| CN101050435A (en) | Solid microbe agent for degrading petroleum pollution, and petroleum products, and preparation method | |
| CN103103142B (en) | Staphylococcus cohnii and applications thereof | |
| US8163534B2 (en) | Estrogenic substance degradable microorganism and use thereof | |
| CN112725240B (en) | Acinetobacter livelii, and microbial inoculum and application thereof | |
| CN107299066B (en) | Preparation method and degradation treatment method of microbial degradation liquid containing oily sludge | |
| CN108277175A (en) | 2,4 dinitrotoluene (DNT) sulfonate efficient degrading bacterial strain Microbacterium sp.X3 and its application | |
| KR20000034035A (en) | Bioaugmentation of oil contaminated soil by microbial composition which can decompose hydrocarbons derived from petroleum | |
| CN101935631B (en) | Ralstoniasp. and application thereof in bioremediation of petroleum-contaminated saline-alkali soil | |
| CN114854626A (en) | Pseudomonas strain for degrading polycyclic aromatic hydrocarbon pollutants and application thereof | |
| CN103103141B (en) | Kocuria palustris strain and applications thereof | |
| CN117025490A (en) | Strain and microbial inoculum for soil remediation and application thereof | |
| CN106119150A (en) | A kind of dephosphorization microbial bacterial agent and its preparation method and application | |
| CN114717152B (en) | Composite functional microbial agent taking pseudomonas stutzeri as carrier for in-situ remediation of polluted soil and application thereof | |
| CN109554303B (en) | Microbacterium aurantiacus and preparation and application thereof | |
| CN103381418B (en) | Method for processing tobacco waste or organic fluorine wastewater | |
| CN115786179A (en) | Bacterial strain for degrading o-dichlorobenzene and application thereof | |
| CN112646742B (en) | Activated sludge culture agent and application thereof | |
| CN105670965B (en) | Strain with iron reduction capacity and application thereof | |
| CN114262679A (en) | Autotrophic denitrifying bacteria and rapid propagation process thereof | |
| CN106399200A (en) | Alcaligenes sp. and application thereof to high-salt high-polymer waste water | |
| CN111944719A (en) | Dioxane degrading bacterium IS20 as well as preparation method and application thereof | |
| CN107828692B (en) | Terres tarum and preparation and application of microbial agent thereof | |
| CN101074423A (en) | Globular bacillus and its use | |
| CN109666613A (en) | A kind of facultative autotrophy has rhizobium and its application of nitrate reduction ferrous oxidation function |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |