CN117535195A - Composite microbial agent for degrading antibiotics and pesticide residues and preparation method thereof - Google Patents
Composite microbial agent for degrading antibiotics and pesticide residues and preparation method thereof Download PDFInfo
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- CN117535195A CN117535195A CN202311549353.2A CN202311549353A CN117535195A CN 117535195 A CN117535195 A CN 117535195A CN 202311549353 A CN202311549353 A CN 202311549353A CN 117535195 A CN117535195 A CN 117535195A
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 67
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 66
- 239000002131 composite material Substances 0.000 title claims abstract description 51
- 239000003242 anti bacterial agent Substances 0.000 title claims abstract description 43
- 229940088710 antibiotic agent Drugs 0.000 title claims abstract description 43
- 239000000447 pesticide residue Substances 0.000 title claims abstract description 36
- 230000000593 degrading effect Effects 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 239000000575 pesticide Substances 0.000 claims abstract description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 24
- 229910001868 water Inorganic materials 0.000 claims abstract description 24
- 239000002689 soil Substances 0.000 claims abstract description 20
- 150000001875 compounds Chemical class 0.000 claims abstract description 18
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 14
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 14
- 241000881860 Paenibacillus mucilaginosus Species 0.000 claims abstract description 14
- 239000007633 bacillus mucilaginosus Substances 0.000 claims abstract description 14
- 241000563903 Bacillus velezensis Species 0.000 claims abstract description 10
- 241001673062 Achromobacter xylosoxidans Species 0.000 claims abstract description 9
- 241000588624 Acinetobacter calcoaceticus Species 0.000 claims abstract description 9
- 241000194105 Paenibacillus polymyxa Species 0.000 claims abstract description 9
- 241001460073 Trichoderma asperellum Species 0.000 claims abstract description 9
- 238000002156 mixing Methods 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 25
- 230000001580 bacterial effect Effects 0.000 claims description 20
- 239000001963 growth medium Substances 0.000 claims description 9
- 239000002068 microbial inoculum Substances 0.000 claims description 9
- 244000005700 microbiome Species 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 235000015278 beef Nutrition 0.000 claims description 6
- 230000003115 biocidal effect Effects 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 claims description 4
- 239000000152 carbamate pesticide Substances 0.000 claims description 4
- 238000005507 spraying Methods 0.000 claims description 4
- 150000003852 triazoles Chemical class 0.000 claims description 4
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 3
- 238000009630 liquid culture Methods 0.000 claims description 3
- 238000012807 shake-flask culturing Methods 0.000 claims description 3
- 241000251468 Actinopterygii Species 0.000 claims description 2
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 2
- 239000005822 Propiconazole Substances 0.000 claims description 2
- 239000011717 all-trans-retinol Substances 0.000 claims description 2
- 235000019169 all-trans-retinol Nutrition 0.000 claims description 2
- 239000006013 carbendazim Substances 0.000 claims description 2
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 2
- DUEPRVBVGDRKAG-UHFFFAOYSA-N carbofuran Chemical compound CNC(=O)OC1=CC=CC2=C1OC(C)(C)C2 DUEPRVBVGDRKAG-UHFFFAOYSA-N 0.000 claims description 2
- JLQUFIHWVLZVTJ-UHFFFAOYSA-N carbosulfan Chemical compound CCCCN(CCCC)SN(C)C(=O)OC1=CC=CC2=C1OC(C)(C)C2 JLQUFIHWVLZVTJ-UHFFFAOYSA-N 0.000 claims description 2
- CWFOCCVIPCEQCK-UHFFFAOYSA-N chlorfenapyr Chemical compound BrC1=C(C(F)(F)F)N(COCC)C(C=2C=CC(Cl)=CC=2)=C1C#N CWFOCCVIPCEQCK-UHFFFAOYSA-N 0.000 claims description 2
- 229910052801 chlorine Inorganic materials 0.000 claims description 2
- 239000000460 chlorine Substances 0.000 claims description 2
- RDYMFSUJUZBWLH-SVWSLYAFSA-N endosulfan Chemical compound C([C@@H]12)OS(=O)OC[C@@H]1[C@]1(Cl)C(Cl)=C(Cl)[C@@]2(Cl)C1(Cl)Cl RDYMFSUJUZBWLH-SVWSLYAFSA-N 0.000 claims description 2
- 239000003993 organochlorine pesticide Substances 0.000 claims description 2
- STJLVHWMYQXCPB-UHFFFAOYSA-N propiconazole Chemical compound O1C(CCC)COC1(C=1C(=CC(Cl)=CC=1)Cl)CN1N=CN=C1 STJLVHWMYQXCPB-UHFFFAOYSA-N 0.000 claims description 2
- 229940072172 tetracycline antibiotic Drugs 0.000 claims description 2
- FFSJPOPLSWBGQY-UHFFFAOYSA-N triazol-4-one Chemical compound O=C1C=NN=N1 FFSJPOPLSWBGQY-UHFFFAOYSA-N 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 3
- 238000013329 compounding Methods 0.000 abstract description 3
- 230000000052 comparative effect Effects 0.000 description 26
- 244000144972 livestock Species 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 230000015556 catabolic process Effects 0.000 description 7
- 238000006731 degradation reaction Methods 0.000 description 7
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 244000144977 poultry Species 0.000 description 4
- 239000012752 auxiliary agent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 210000003608 fece Anatomy 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 239000010871 livestock manure Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 239000004098 Tetracycline Substances 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 235000019364 tetracycline Nutrition 0.000 description 2
- 150000003522 tetracyclines Chemical class 0.000 description 2
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- 229960004475 chlortetracycline Drugs 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- 238000009264 composting Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 239000004021 humic acid Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000003120 macrolide antibiotic agent Substances 0.000 description 1
- 229940041033 macrolides Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000007660 quinolones Chemical class 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000005067 remediation Methods 0.000 description 1
- 238000003900 soil pollution Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
-
- 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
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
- B09C1/105—Reclamation of contaminated soil microbiologically, biologically or by using enzymes using fungi or plants
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/306—Pesticides
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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
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- C12R2001/05—Alcaligenes
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- C12R2001/12—Bacillus polymyxa ; Paenibacillus polymyxa
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- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- C12R2001/885—Trichoderma
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Abstract
The invention belongs to the field of microbial preparations, and discloses a composite microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof, wherein the composite microbial agent comprises the following main components: bacillus subtilis, bacillus methylotrophicus, acinetobacter calcoaceticus, trichoderma asperellum, paenibacillus polymyxa, bacillus mucilaginosus, and alcaligenes xylosoxidans; the preparation method is that after the corresponding various bacteria are cultured independently, the bacteria with the corresponding concentration are obtained, and the composite microbial agent is obtained after mixing. The invention can effectively degrade various antibiotics and residual pesticides by compounding various strains, and can effectively improve the soil and water body polluted by the antibiotics and the pesticides. The invention is suitable for preparing the compound microbial agent for degrading various antibiotics and residual pesticides, and is used for improving the soil and water body polluted by the antibiotics and the residual pesticides.
Description
Technical Field
The invention belongs to the field of microbial agents, and relates to a composite microbial agent, in particular to a composite microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof.
Background
In order to make the crops and livestock grow better, the growth vigor of the crops and livestock is promoted, and meanwhile, the diseases of the crops and the livestock are controlled. The common method is to use pesticides and various antibiotics, and although the pesticides and the antibiotics can promote the healthy growth of crops and livestock, the used pesticides and the antibiotics cannot be fully utilized by the crops and the livestock and enter soil and water bodies through various ways, so that the soil and the water bodies are polluted and the human health is endangered.
About 30 to 90 percent of antibiotics in the livestock and poultry breeding industry are not absorbed and utilized by animal internal organs, and are taken as raw medicines or
The metabolite form of the polypeptide is discharged out of the body through the feces and urine. Comprehensively analyzing the detection condition of livestock and poultry manure antibiotics at home and abroad, finding tetracyclines, sulfonamides, quinolones, macrolides and the like as antibiotics widely used in the livestock and poultry industry at present, and the highest detection concentration of the antibiotics can reach tens or even hundreds of mg/kg. In recent years, the investigation work of livestock and poultry manure antibiotics is gradually increased, and the highest detection concentration of the tetracycline aureomycin reaches 764.4mg/kg. Composting and anaerobic digestion are the main modes of harmless and recycling treatment of livestock manure at present.
In terms of pesticide residues, however, the pesticide residues pose a hazard to the ecological environment and human health. Biodegradation is an effective way for removing pesticide residues, and has the advantages of high efficiency, low cost, safety and the like compared with the traditional physical and chemical methods, and has been paid attention to in recent years.
The chemical pesticide has the advantages of quick effect, wide control spectrum, stable property, convenient storage and transportation, low price and the like, plays a great role in controlling crop diseases, insect pests and diseases, and the like, but the use of a large amount of pesticide also brings side effects of environmental pollution, killing natural enemies, generating drug resistance and the like, causing adverse effects of increased pesticide residues, reduced land fertility, reduced quality of agricultural products and the like, and particularly brings serious threat to human health because of long pesticide residue period with high toxicity and stable property and difficult degradation.
In the remediation of contaminated soil, bioremediation is attracting attention due to low treatment cost and no secondary pollution. In recent years, the application of microorganisms in agriculture is wider, and the microorganisms play an important role in degrading pesticide residues. The degradation of microorganisms is the most important and thorough purification of pesticides in the soil. Microorganisms having a degrading ability to pesticides include bacteria, fungi, etc. The bioremediation method for degrading pollutants by utilizing organisms or biological products has the advantages of no toxicity, no residue and the like, is a relatively cheap and effective method for eliminating pesticide residues or reducing the toxicity of the residual pesticide, and shows the inhibition effect on crop diseases and insect pests to different degrees when the microbial inoculum is used as a fertilizer.
Microorganisms are very abundant in metabolic types due to various kinds, and can thoroughly decompose pollutants into substances harmless to the environment and human bodies, such as carbon dioxide, water, inorganic compounds and the like. In addition, the microbial degradation pesticide has substrate specificity, and can only degrade one or a class of pesticides with similar structures, so that the repair cost for the pollution of the composite pesticide can be increased. On the other hand, the shelf life of the degradation microbial inoculum product is prolonged, and the stability of the field degradation and repair effect is further improved.
Therefore, it is necessary to provide a compound microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims to provide a composite microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof, so as to achieve the purpose of simultaneously degrading various antibiotics and pesticide residues in soil and/or water.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a compound microbial agent for degrading antibiotics and pesticide residues comprises the following main components:
bacillus subtilis, bacillus methylotrophicus, acinetobacter calcoaceticus, trichoderma asperellum, paenibacillus polymyxa, bacillus mucilaginosus, and alcaligenes xylosoxidans; the volume ratio of the bacterial liquid of each bacterial strain is 1-4:3-7:1-8:5-11:2-5:3-6:4-16, and the effective viable count in each bacterial liquid is more than or equal to 0.2 x 10 8 cfu/g；
The bacillus subtilis is ATCC6633, the bacillus methylotrophicus is CGMCC 1.12059, the acinetobacter calcoaceticus is ATCC23055, the trichoderma asperellum is TaspHu1, the paenibacillus polymyxa is CGMCC 7250, the bacillus mucilaginosus is CGMCC1.231, the alcaligenes xylosoxidans is ATCC27061, and the above strains can be purchased through conventional commercial channels;
preferably, the volume ratio of the strain liquid is 2:5:7:5:3:4:9;
as the limit of the invention, the total effective viable count of the compound microorganism bacterial agent is 1.24 x 10 9 ～5.56*10 10 cfu/g；
The effective viable count in each bacterial liquid is as follows:
bacillus subtilis: 3.0*10 8 ～4.0*10 9 cfu/g；
Methyl nutrition typeBacillus: 2.0*10 8 ～1.0*10 10 cfu/g；
Acinetobacter calcoaceticus: 4.0*10 8 ～2.0*10 10 cfu/g；
Trichoderma asperellum: 0.8*10 8 ～1.0*10 10 cfu/g；
Paenibacillus polymyxa: 2.0*10 8 ～6.0*10 9 cfu/g；
Bacillus mucilaginosus: 0.2*10 8 ～4.0*10 9 cfu/g；
Alcaligenes xylosoxidans: 0.4*10 8 ～1.6*10 9 cfu/g；
As another definition of the invention, the antibiotic is a tetracycline antibiotic;
as a third limitation of the present invention, the pesticide is at least one of a carbamate pesticide, an organochlorine pesticide, and a triazole pesticide;
as a further limitation of the present invention, the carbamate pesticide is at least one of carbendazim, carbosulfan, zineb, and furadan;
the organic chlorine pesticide is at least one of endosulfan and chlorfenapyr;
the triazole pesticide is at least one of triazolone and propiconazole
The invention also provides a preparation method of the composite microbial agent for degrading antibiotics and pesticide residues, which comprises the following steps:
(1) Activating strains: respectively inoculating various strains stored on the inclined plane to a beef extract peptone culture medium plate, and culturing for 24-36 h at the temperature of 30-32 ℃;
(2) Seed liquid culture: respectively picking the activated strains, inoculating the activated strains into a liquid beef extract peptone culture medium, and shake-flask culturing for 24-36 h at the temperature of 30-32 ℃;
(3) And (3) performing expansion culture: inoculating the seed liquid into a liquid fermentation culture medium according to the inoculum size of 1-10%, culturing for 12-24 h at 30-32 ℃, rotating at 180-260 rpm, and ventilating at 0.6-1.2V/V.min;
(4) Preparation of a microbial inoculum: respectively collecting the needed bacteriaThe bacterial strain is prepared into bacterial liquid, the bacterial liquid of each bacterial strain is mixed according to the volume ratio of 1-4:3-7:1-8:5-11:2-5:3-6:4-16, and the total effective viable count is 5.0 x 10 8 ～1.0*10 11 cfu/g of composite microbial agent;
the invention also provides application of the composite microbial agent for degrading antibiotics and pesticide residues, which is mainly applied to soil treatment or crop treatment;
the soil treatment is as follows: before ploughing, uniformly spraying the composite microbial agent on the ground, and then opening ridges and ploughing the ground;
the crop treatment is as follows: spraying the overground part of the crops in the growing period of the crops by using the composite microbial agent, and irrigating roots of the underground crops in the growing period;
the water treatment is as follows: before being put into fish fry, the volume ratio of the water body to the composite microbial agent is 13-24: 1, uniformly scattering the composite microorganism into the water body, and standing the water body for 24-72 h.
By adopting the technical scheme, compared with the prior art, the invention has the following beneficial effects:
(1) According to the composite microbial agent for degrading the antibiotics and the pesticide residues, 7 strains are selected, so that the strains can coexist well, a high-efficiency metabolic degradation system can be formed, and compared with a single strain, the efficiency of degrading the pesticides and the antibiotics can be obviously improved;
(2) The composite microbial agent for degrading antibiotics and pesticide residues provided by the invention can degrade antibiotics and pesticide residues in soil, and can degrade pesticide residues in water, improve water quality and facilitate storage.
In conclusion, the invention can effectively degrade various antibiotics and residual pesticides through the compounding of various strains, and can effectively improve the soil and water body polluted by the antibiotics and the pesticides.
The invention is suitable for preparing the compound microbial agent for degrading various antibiotics and residual pesticides, and is used for improving the soil and water body polluted by the antibiotics and the residual pesticides.
Detailed Description
Preferred embodiments of the present invention are described below. It should be understood that the preferred embodiments described herein are presented for purposes of illustration and understanding only, and are not intended to limit the invention.
Example 1A composite microbial agent for degrading antibiotic and pesticide residues
The embodiment provides a compound microbial agent for degrading antibiotics and pesticide residues, which comprises the following main strains: bacillus subtilis, methylotrophic bacillus, acinetobacter calcoaceticus, trichoderma asperellum, paenibacillus polymyxa, bacillus mucilaginosus and Alcaligenes xylosoxidans, wherein the volume ratio of the strains is 2:5:7:5:3:4:9
The total effective viable count of the composite microbial agent is 1.24 x 10 9 cfu/g；
The effective viable count in each bacterial liquid is as follows:
bacillus subtilis: 3.0*10 8 cfu/g；
Methylotrophic bacillus: 2.0*10 8 cfu/g；
Acinetobacter calcoaceticus: 4.0*10 8 cfu/g；
Trichoderma asperellum: 0.8*10 8 cfu/g；
Paenibacillus polymyxa: 2.0*10 8 cfu/g；
Bacillus mucilaginosus: 0.2*10 8 cfu/g；
Alcaligenes xylosoxidans: 0.4*10 8 cfu/g；
The formula of the microbial inoculum also comprises an auxiliary agent, wherein the main components of the microbial inoculum comprise 7 parts of diatomite, 2 parts of humic acid and 1 part of ammonium sulfate by weight;
the preparation method of the microbial inoculum comprises the following steps:
(1) Activating strains: respectively inoculating various strains stored on the inclined plane to a beef extract peptone culture medium plate, and culturing for 24-36 h at the temperature of 30-32 ℃;
(2) Seed liquid culture: respectively picking the activated strains, inoculating the activated strains into a liquid beef extract peptone culture medium, and shake-flask culturing for 24-36 h at the temperature of 30-32 ℃;
(3) And (3) performing expansion culture: inoculating the seed liquid into a liquid fermentation culture medium according to the inoculum size of 1-10%, culturing for 12-24 h at 30-32 ℃, rotating at 180-260 rpm, and ventilating at 0.6-1.2V/V.min;
(4) Preparation of a microbial inoculum: respectively collecting thallus of required strains to prepare bacterial liquid, mixing bacterial liquid of each strain with prepared auxiliary agent according to the volume ratio of 2:5:7:5:3:4:9 to obtain the total effective viable count of 1.24 x 10 9 ～5.56*10 10 cfu/g of composite microbial agent.
EXAMPLES 2-6 Compound microbial agent for degrading antibiotics and pesticide residues
Examples 2 to 6 respectively provide a compound microbial agent for degrading antibiotics and pesticide residues, the main components and the preparation method of the compound microbial agent are the same as those of example 1, the difference is only that the volume ratio of each strain, the total effective viable count of the compound microbial agent and the effective viable count in each bacterial liquid are shown in table 1.
Table 1:
comparative example 1
The composite microbial agent of the comparative example 1 and the preparation method thereof are the same as those of the example 1, except that no auxiliary agent is added in the comparative example;
comparative example 2
The composite microbial agent of this comparative example 2 and the preparation method thereof are the same as those of example 1, except that no Bacillus subtilis was added in this comparative example;
comparative example 3
The compound microbial agent of this comparative example 3 and the preparation method thereof are the same as those of example 1, except that Bacillus methylotrophicus is not added in this comparative example;
comparative example 4
The composite microbial agent of comparative example 4 and the preparation method thereof are the same as those of example 1, except that Bacillus mucilaginosus is not added in the comparative example;
comparative example 5
The composite microbial agent of this comparative example 5 and the method for producing the same are the same as in example 1, except that this comparative example was not added with Bacillus subtilis and Bacillus methylotrophicus;
comparative example 6
The composite microbial agent of this comparative example 6 and the preparation method thereof are the same as those of example 1, except that the present comparative example is free of Bacillus subtilis and Bacillus mucilaginosus;
comparative example 7
The composite microbial agent of comparative example 7 and the preparation method thereof are the same as those of example 1, except that Bacillus mucilaginosus and Bacillus methylotrophicus are not added in this comparative example;
comparative example 8
The composite microbial agent of this comparative example 8 and the preparation method thereof are the same as in example 1, except that bacillus subtilis, bacillus mucilaginosus and bacillus methylotrophicus are not added in this comparative example;
test examples
According to the actual soil and water pollution environments, 15 groups of polluted soil and water models mixed with antibiotics and residual pesticides are respectively established, the polluted soil and water are correspondingly treated by using the composite microbial agents related to examples 1-6 and comparative examples 1-8, and the rest 1 groups of polluted soil and water environments are taken as blank comparative examples, and the specific test results are shown in table 2.
Table 2:
the test result shows that the compound microbial agent obtained by compounding according to the volume ratio in the embodiment 1 has the best effect on degrading antibiotics and pesticide residues in soil and water, and simultaneously shows that the effect of the compound microbial agent is obtained by the mutual coordination of three strains, namely bacillus subtilis, bacillus mucilaginosus and bacillus methylotrophicus; the three strains can mutually promote and strengthen the degradation effect on antibiotics and pesticides in soil and water.
The foregoing describes one embodiment of the present invention in detail, but the description is only a preferred embodiment of the present invention and should not be construed as limiting the scope of the invention. All equivalent changes and modifications within the scope of the present invention are intended to be covered by the present invention.
Claims (10)
1. A compound microbial agent for degrading antibiotics and pesticide residues is characterized in that: the main components of the compound microbial agent comprise:
bacillus subtilis, bacillus methylotrophicus, acinetobacter calcoaceticus, trichoderma asperellum, paenibacillus polymyxa, bacillus mucilaginosus, and alcaligenes xylosoxidans; the volume ratio of the bacterial liquid of each bacterial strain is 1-4:3-7:1-8:5-11:2-5:3-6:4-16, and the effective viable count in each bacterial liquid is more than or equal to 0.2 x 10 8 cfu/g。
2. The composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the composite microbial agent is characterized in that: the main components of the compound microbial agent comprise:
bacillus subtilis, bacillus methylotrophicus, acinetobacter calcoaceticus, trichoderma asperellum, paenibacillus polymyxa, bacillus mucilaginosus and alcaligenes xylosoxidans, wherein the volume ratio of bacterial liquid of each strain is 2:5:7:5:3:4:9.
3. The composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the composite microbial agent is characterized in that: the total effective viable count of the compound microbial agent is 1.24 x 10 9 ～5.56*10 10 cfu/g; the effective viable count in each bacterial liquid is as follows:
bacillus subtilis: 3.0*10 8 ～4.0*10 9 cfu/g；
Methylotrophic bacillus: 2.0*10 8 ～1.0*10 10 cfu/g；
Acinetobacter calcoaceticus: 4.0*10 8 ～2.0*10 10 cfu/g；
Trichoderma asperellum: 0.8*10 8 ～1.0*10 10 cfu/g；
Paenibacillus polymyxa: 2.0*10 8 ～6.0*10 9 cfu/g；
Bacillus mucilaginosus: 0.2*10 8 ～4.0*10 9 cfu/g；
Alcaligenes xylosoxidans: 0.4*10 8 ～1.6*10 9 cfu/g。
4. The composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the composite microbial agent is characterized in that: the antibiotic is tetracycline antibiotics.
5. The composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the composite microbial agent is characterized in that: the pesticide is at least one of carbamate pesticide, organochlorine pesticide and triazole pesticide.
6. The composite microbial agent for degrading antibiotic and pesticide residues according to claim 5, wherein the composite microbial agent is characterized in that: the carbamate pesticide is at least one of carbendazim, carbosulfan, zineb and furadan.
7. The composite microbial agent for degrading antibiotic and pesticide residues according to claim 5, wherein the composite microbial agent is characterized in that: the organic chlorine pesticide is at least one of endosulfan and chlorfenapyr.
8. The composite microbial agent for degrading antibiotic and pesticide residues according to claim 5, wherein the composite microbial agent is characterized in that: the triazole pesticide is at least one of triazolone and propiconazole.
9. The method for preparing the composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the method is characterized by comprising the following steps: the method comprises the following steps:
(1) Activating strains: respectively inoculating various strains stored on the inclined plane to a beef extract peptone culture medium plate, and culturing for 24-36 h at the temperature of 30-32 ℃;
(2) Seed liquid culture: respectively picking the activated strains, inoculating the activated strains into a liquid beef extract peptone culture medium, and shake-flask culturing for 24-36 h at the temperature of 30-32 ℃;
(3) And (3) performing expansion culture: inoculating the seed liquid into a liquid fermentation culture medium according to the inoculum size of 1-10%, culturing for 12-24 h at 30-32 ℃, rotating at 180-260 rpm, and ventilating at 0.6-1.2V/V.min;
(4) Preparation of a microbial inoculum: respectively collecting thallus of required strains to prepare bacterial liquid, mixing bacterial liquid of each strain according to the volume ratio of 1-4:3-7:1-8:5-11:2-5:3-6:4-16 to obtain the total effective viable count of 5.0 x 10 8 ～1.0*10 11 cfu/g of composite microbial agent.
10. The use of the composite microbial agent for degrading antibiotics and pesticide residues according to claim 1, wherein the composite microbial agent is characterized in that: the compound microbial agent is used for soil treatment, crop treatment or water treatment;
the soil treatment is as follows: before ploughing, uniformly spraying the composite microbial agent on the ground, and then opening ridges and ploughing the ground;
the crop treatment is as follows: spraying the overground part of the crops in the growing period of the crops by using the composite microbial agent, and irrigating roots of the underground crops in the growing period;
the water treatment is as follows: before being put into fish fry, the composite microorganism is uniformly scattered into the water body according to the volume ratio of the water body to the composite microorganism microbial inoculum of 13-24:1, and the water body is kept stand for 24-72 h.
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