CN116286552A - Compound microbial agent for degrading antibiotics and pesticide residues and preparation method thereof - Google Patents
Compound microbial agent for degrading antibiotics and pesticide residues and preparation method thereof Download PDFInfo
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- CN116286552A CN116286552A CN202310460036.7A CN202310460036A CN116286552A CN 116286552 A CN116286552 A CN 116286552A CN 202310460036 A CN202310460036 A CN 202310460036A CN 116286552 A CN116286552 A CN 116286552A
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- microbial agent
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- bacterial liquid
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 65
- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 64
- 239000003242 anti bacterial agent Substances 0.000 title claims abstract description 27
- 229940088710 antibiotic agent Drugs 0.000 title claims abstract description 27
- 239000000447 pesticide residue Substances 0.000 title claims abstract description 17
- 230000000593 degrading effect Effects 0.000 title claims abstract description 16
- 150000001875 compounds Chemical class 0.000 title abstract description 22
- 238000002360 preparation method Methods 0.000 title abstract description 11
- 239000002131 composite material Substances 0.000 claims abstract description 39
- 230000001580 bacterial effect Effects 0.000 claims abstract description 30
- 239000007788 liquid Substances 0.000 claims abstract description 27
- 239000000575 pesticide Substances 0.000 claims abstract description 24
- 241001673062 Achromobacter xylosoxidans Species 0.000 claims abstract description 22
- 241000588624 Acinetobacter calcoaceticus Species 0.000 claims abstract description 21
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 21
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 21
- 241000193388 Bacillus thuringiensis Species 0.000 claims abstract description 21
- 241001460073 Trichoderma asperellum Species 0.000 claims abstract description 21
- 229940097012 bacillus thuringiensis Drugs 0.000 claims abstract description 21
- 241000186046 Actinomyces Species 0.000 claims abstract description 19
- 239000002689 soil Substances 0.000 claims abstract description 14
- 241000894006 Bacteria Species 0.000 claims abstract description 10
- 150000003852 triazoles Chemical class 0.000 claims abstract description 7
- 239000003993 organochlorine pesticide Substances 0.000 claims abstract description 6
- 229940072172 tetracycline antibiotic Drugs 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 11
- 230000003115 biocidal effect Effects 0.000 claims description 9
- 238000005507 spraying Methods 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
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 claims description 3
- 239000006013 carbendazim Substances 0.000 claims description 3
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 claims description 3
- 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 3
- 238000003973 irrigation Methods 0.000 claims description 3
- 230000002262 irrigation Effects 0.000 claims description 3
- FFSJPOPLSWBGQY-UHFFFAOYSA-N triazol-4-one Chemical compound O=C1C=NN=N1 FFSJPOPLSWBGQY-UHFFFAOYSA-N 0.000 claims description 3
- 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
- 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
- 229960001952 metrifonate Drugs 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
- NFACJZMKEDPNKN-UHFFFAOYSA-N trichlorfon Chemical compound COP(=O)(OC)C(O)C(Cl)(Cl)Cl NFACJZMKEDPNKN-UHFFFAOYSA-N 0.000 claims description 2
- 230000015556 catabolic process Effects 0.000 abstract description 18
- 238000006731 degradation reaction Methods 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 5
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 abstract description 3
- 244000144972 livestock Species 0.000 description 9
- 238000009264 composting Methods 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
- 230000029087 digestion Effects 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 238000002156 mixing Methods 0.000 description 7
- 239000004098 Tetracycline Substances 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 210000003608 fece Anatomy 0.000 description 5
- 239000010871 livestock manure Substances 0.000 description 5
- 244000144977 poultry Species 0.000 description 5
- 235000019364 tetracycline Nutrition 0.000 description 5
- 150000003522 tetracyclines Chemical class 0.000 description 5
- 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 4
- 229960004475 chlortetracycline Drugs 0.000 description 4
- 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 4
- 235000019365 chlortetracycline Nutrition 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 229960002180 tetracycline Drugs 0.000 description 4
- 229930101283 tetracycline Natural products 0.000 description 4
- 239000004100 Oxytetracycline Substances 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 229960000625 oxytetracycline Drugs 0.000 description 3
- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[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 IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 description 3
- 235000019366 oxytetracycline Nutrition 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 description 3
- 241000186361 Actinobacteria <class> Species 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- YASYVMFAVPKPKE-UHFFFAOYSA-N acephate Chemical compound COP(=O)(SC)NC(C)=O YASYVMFAVPKPKE-UHFFFAOYSA-N 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 238000004064 recycling Methods 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000002154 agricultural waste Substances 0.000 description 1
- 239000003674 animal food additive Substances 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
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
- 239000002054 inoculum Substances 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
- 230000007246 mechanism Effects 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- NNKVPIKMPCQWCG-UHFFFAOYSA-N methamidophos Chemical class COP(N)(=O)SC NNKVPIKMPCQWCG-UHFFFAOYSA-N 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000001089 mineralizing effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000009304 pastoral farming Methods 0.000 description 1
- 238000009374 poultry farming Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 150000007660 quinolones Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 229940124530 sulfonamide Drugs 0.000 description 1
- 150000003456 sulfonamides Chemical class 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 229940040944 tetracyclines Drugs 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/32—Ingredients for reducing the noxious effect of the active substances to organisms other than pests, e.g. toxicity reducing compositions, self-destructing compositions
-
- 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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- 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
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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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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/05—Alcaligenes
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- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/075—Bacillus thuringiensis
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- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- C12R2001/885—Trichoderma
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention relates to the technical field of microbial agents, in particular to a composite for degrading antibiotics and pesticide residuesA microbial agent and its preparation method are provided. The application discloses a compound microbial agent for degrading antibiotics and pesticide residues, wherein the microbial agent consists of the following bacteria: bacillus subtilis, bacillus thuringiensis, acinetobacter calcoaceticus, trichoderma asperellum, actinomyces and alcaligenes xylosoxidans; the volume ratio of the bacterial liquid of each bacterial strain is 1:1:1:1:1:1, 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 thuringiensis is ATCC10792, the Acinetobacter calcoaceticus is ATCC23055, the trichoderma asperellum is TaspHu1, the actinomyces is ATCC27044, and the alcaligenes xylosoxidans is ATCC27061. The composite microbial agent prepared by the application has obvious degradation effect on carbamate pesticides, organochlorine pesticides, triazole pesticides and tetracycline antibiotics in soil.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to a composite microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof.
Background
With the continuous development of intensive livestock breeding industry, antibiotics are widely used as feed additives to control livestock diseases and promote the growth of livestock. Most antibiotics used in livestock and poultry raising industry cannot be completely absorbed by a passive body, but are discharged out of the body along with animal manure in the form of original shape or metabolites, and then enter soil and water body through various ways, so that great threat is brought to human health.
About 30-90% of antibiotics in livestock and poultry farming are not absorbed and utilized by animal internal organs, and are discharged outside the body through feces and urine in the form of raw medicines or metabolites thereof. 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.
1. Composting process
Composting refers to the process of mineralizing, humifying and innocuously converting organic matters into decomposed fertilizers by utilizing the decomposition effect of microorganisms under the high-temperature condition. Composting is generally classified into aerobic composting and anaerobic composting. The aerobic composting has the characteristics of high temperature, capability of better killing pathogenic microorganisms, short composting time, high efficiency and the like, and is widely applied to the treatment of livestock and poultry manure;
2. anaerobic digestion treatment
Anaerobic digestion refers to the process of finally producing carbon dioxide and methane under the action of microbial decomposition and conversion of organic matters under anaerobic conditions. Anaerobic digestion can be classified into: low temperature anaerobic digestion (below 25 ℃), medium temperature anaerobic digestion (30-45 ℃) and high temperature anaerobic digestion (50-65 ℃). The anaerobic digestion technology is widely used for recycling industrial and agricultural wastes due to the advantages of capability of treating high-concentration organic wastewater, easiness in operation, capability of generating clean energy and the like.
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. For example, most pesticide microorganism degradation metabolism mechanisms including acephate are not completely resolved at present, pure culture microorganisms obtained through screening cannot completely degrade the acephate, and intermediate metabolites of methamidophos with stronger toxicity are generated, so that development and utilization of degradation microorganisms are greatly limited.
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 overcome the defects of the prior art, the invention aims to provide a composite microbial agent for degrading antibiotics and pesticide residues and a preparation method thereof.
The technical scheme adopted for solving the technical problems is as follows:
the first object of the invention is to provide a compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis, bacillus thuringiensis, acinetobacter calcoaceticus, trichoderma asperellum, actinomyces and alcaligenes xylosoxidans;
the bacillus subtilis is ATCC6633, the bacillus thuringiensis is ATCC10792, the Acinetobacter calcoaceticus is ATCC23055, the trichoderma asperellum is TaspHu1, the actinomyces is ATCC27044, and the alcaligenes xylosoxidans is ATCC27061. The effective viable count in each bacterial liquid is more than or equal to 0.2 x 108cfu/g;
further, the total effective viable count of the compound microbial agent is 4.0 x 10 8 -2.0*10 10 cfu/g; the effective viable bacteria of each strain are composed of the following components:
further, the antibiotic is a tetracycline antibiotic. Specifically, one or more of oxytetracycline, tetracycline and aureomycin can be selected.
Further, the pesticide is one or more of carbamate pesticide, organochlorine pesticide and triazole pesticide.
Further, the carbamate pesticide is one or more of carbendazim, carbosulfan, zineb and furadan.
Further, the organic chlorine pesticide is one or two of endosulfan and trichlorfon.
Furthermore, the triazole pesticide is one or two of triazolone and propiconazole.
The second object of the present invention is to provide a method for preparing the composite microbial agent, wherein the corresponding bacteria are independently culturedThen obtaining strains with corresponding concentration, mixing to obtain the effective viable count of 4.0 x 10 8 -2.0*10 10 cfu/g of composite microbial agent.
The third object of the present invention is to provide a method for using the composite microbial agent, wherein the composite microbial agent is used for soil treatment or crop treatment. Further, 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;
still further, the crop treatment is: the composite microbial agent is used for spraying the overground part of crops in the growing period of the crops, and the root irrigation treatment can be carried out on underground crops in the growing period.
The beneficial effects of the invention are as follows:
the composite microbial agent is prepared by mixing bacillus subtilis, bacillus thuringiensis, acinetobacter calcoaceticus, trichoderma asperellum, actinomycetes and alcaligenes xylosoxidans, and 6 strains are selected, so that the strains can coexist better, a high-efficiency metabolic degradation system can be formed, and compared with a single strain, the efficiency of degrading pesticides and antibiotics can be obviously improved; the test results show that the compound microbial agent has obvious degradation effect on carbamate pesticides, organochlorine pesticides, triazole pesticides and tetracycline antibiotics in soil.
Detailed Description
The invention will be further illustrated by the following examples, which are not intended to limit the scope of the invention, in order to facilitate the understanding of those skilled in the art.
The purchasing of part of raw materials in the application is as follows: bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomycetes ATCC27044 and Alcaligenes xylosoxidans ATCC27061 are all commercial strains. Namely, the bacillus subtilis/bacillus thuringiensis/Acinetobacter calcoaceticus/actinomyces/trichoderma asperellum/alcaligenes xylosoxidans belong to known strains and can be purchased from commercial paths such as CGMCC, ATCC and the like. Seed culture and fermenter culture of each strain of the present application are conventional culture methods in the art;
the fermentation process parameters of bacillus subtilis, bacillus thuringiensis, acinetobacter calcoaceticus, trichoderma asperellum, actinomyces and alcaligenes xylosoxidans strains are shown in the table 1 below, or the fermentation process parameters can be cultured according to the prior art, so that the bacterial liquid with the effective viable count meeting the requirements can be obtained.
Table 1: different culture conditions of several types of single strains are as follows
The bacillus subtilis ATCC6633, the bacillus thuringiensis ATCC10792, the acinetobacter calcoaceticus ATCC23055, the trichoderma asperellum TaspHu1, the actinomyces ATCC27044 and the alcaligenes xylosoxidans ATCC27061 are respectively and independently cultured, after the thalli grow to the end of a log phase, the bacterial liquid is collected and concentrated, and the concentrated bacterial liquid is uniformly mixed according to the volume ratio of 1.5-2:1.3-1.5:1:1:1.4-1.6:1.2-1.4. Can also be quickly prepared into freeze-dried bacterial powder by freeze-drying technology, the survival rate is 75-80%, and the total effective viable count is more than or equal to 4.0 x 10 8 -2.0*10 10 cfu/g. In the freeze drying technology, the protective agent is skimmed milk. The freeze drying technology is directly adopted in the prior art.
Example 1
A compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and Alcaligenes xylosoxidans ATCC27061.
The total effective viable count of the compound microbial agent is 4.0 x 10 8 cfu/g; the total effective viable count of each strain is as follows:
the preparation method of the composite microbial agent comprises the steps of respectively and independently culturing bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and alcaligenes xylosoxidans ATCC27061, collecting bacterial liquid after the bacterial liquid grows to the end of a log phase, concentrating, and uniformly mixing the concentrated bacterial liquid according to the volume ratio of 1.5:1.3:1:1.4:1.2.
Example 2
A compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and Alcaligenes xylosoxidans ATCC27061.
The total effective viable count of the compound microbial agent is 2.0 x 10 10 cfu/g; the total effective viable count of each strain is as follows:
the preparation method of the composite microbial agent comprises the steps of respectively and independently culturing bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and alcaligenes xylosoxidans ATCC27061, collecting bacterial liquid after the bacterial liquid grows to the end of a log phase, concentrating, and uniformly mixing the concentrated bacterial liquid according to the volume ratio of 2:1.5:1:1.6:1.4.
Example 3
A compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and Alcaligenes xylosoxidans ATCC27061.
The total effective viable count of the compound microbial agent is 1.0 x 10 10 cfu/g; the total effective viable count of each strain is as follows:
alcaligenes xylosoxidans 1.0 x 10 9 cfu/g。
The preparation method of the composite microbial agent comprises the steps of respectively and independently culturing bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and alcaligenes xylosoxidans ATCC27061, collecting bacterial liquid after the bacterial liquid grows to the end of a log phase, concentrating, and uniformly mixing the concentrated bacterial liquid according to the volume ratio of 1.7:1.4:1:1.5:1.3.
Example 4
A compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and Alcaligenes xylosoxidans ATCC27061.
The total effective viable count of the compound microbial agent is 8.0 x 10 8 cfu/g; the total effective viable count of each strain is as follows:
the preparation method of the composite microbial agent comprises the steps of respectively and independently culturing bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and alcaligenes xylosoxidans ATCC27061, collecting bacterial liquid after the bacterial liquid grows to the end of a log phase, concentrating, and uniformly mixing the concentrated bacterial liquid according to the volume ratio of 1.5:1.5:1:1.4:1.4.
Example 5
A compound microbial agent for degrading antibiotics and pesticide residues, which consists of the following bacteria:
bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and Alcaligenes xylosoxidans ATCC27061.
The total effective viable count of the compound microbial agent is 4.0 x 10 9 cfu/g; the total effective viable count of each strain is as follows:
the preparation method of the composite microbial agent comprises the steps of respectively and independently culturing bacillus subtilis ATCC6633, bacillus thuringiensis ATCC10792, acinetobacter calcoaceticus ATCC23055, trichoderma asperellum TaspHu1, actinomyces ATCC27044 and alcaligenes xylosoxidans ATCC27061, collecting bacterial liquid after the bacterial liquid grows to the end of a log phase, concentrating, and uniformly mixing the concentrated bacterial liquid according to the volume ratio of 2:1.3:1:1.6:1.2.
Degradation test:
the composite microbial agent prepared in examples 1 to 5 is applied to soil containing pesticides or antibiotics, and then the pesticide/antibiotic content in the soil is measured, and the degradation rate of the pesticide/antibiotic is calculated.
Pesticide/antibiotic detection method: detection is performed as described in "optimization of the method for detecting six and two drips in food" with reference to Ren Xingquan et al (2018); or other pesticide/antibiotic detection methods in the prior art are adopted for detection.
Degradation rate/% = (pre-degradation content after degradation content)/pre-degradation content 100%
1. Pesticide degradation
In the pesticide degradation test, preparing the compound microbial agent (bacterial liquid) in the embodiment 1-5 according to the inoculation amount of 5%, and diluting the bacterial liquid to the mass concentration of 0.05% in sequence; the treatment area at the test site was 2m 2 Each group of medicines is 10g, uniformly sprayed on the surface of a test ground and turned over; the treatment groups are respectively treated by the compound microbial agent 3 days after the corresponding pesticides are applied, the dosage of the compound microbial agent is 1L per square meter, and the compound microbial agent is turned over every other day. After the compound microbial agent is treated for 3 days, sampling is carried out for detection.
Test results: after 3 days, the concentrations of carbamate pesticides (carbendazim), organochlorine pesticides (endosulfan) and triazole pesticides (triazolone) in the soil are reduced by 75.3-80.1%, 93.5-95.6% and 65.3-69.4%, and the composite microbial agent shows obvious degradation effect of antibiotics.
2. Antibiotic degradation
In the antibiotic degradation test, according to the inoculum size of 5% and 10%, the bacterial liquid is mixed with soil, and the mixture is placed in a constant temperature incubator for standing and light-shielding cultivation for 10 days at 30 ℃, and after the cultivation is finished, a sample is collected, and the antibiotic content is measured.
Test results: the composite microbial agent described in the embodiment 1-5 is inoculated in soil, so that the degradation of various antibiotics can be effectively promoted, the concentrations of oxytetracycline, tetracycline and aureomycin in 5% of the inoculation treatment group are reduced by 44.3-45.5%, 27.6-31.2%, 86.8-89.2%, the concentrations of oxytetracycline, tetracycline and aureomycin in 10% of the inoculation treatment group are reduced by 50.1-52.3%, 32.4-34.6% and 90.3-92.5%, and the composite microbial agent shows obvious degradation effects of the antibiotics.
Therefore, in the specific use of the composite microbial agents prepared in examples 1 to 5, the use method may be: the composite microbial agent is used for soil treatment or crop treatment. The soil treatment is as follows: before ploughing, uniformly spraying a composite microbial agent (the mass concentration is 0.05 percent in sequence) on the ground, and then opening ridges and ploughing the ground; the crop treatment is as follows: the composite microbial agent is used for spraying the overground part of crops in the growing period of the crops, and the root irrigation treatment can be carried out on underground crops in the growing period.
The above embodiments are preferred embodiments of the present invention, and besides, the present invention may be implemented in other ways, and any obvious substitution is within the scope of the present invention without departing from the concept of the present invention.
Claims (10)
1. The composite microbial agent for degrading antibiotics and pesticide residues is characterized by comprising the following bacteria:
bacillus subtilis, bacillus thuringiensis, acinetobacter calcoaceticus, trichoderma asperellum, actinomyces and alcaligenes xylosoxidans; the volume ratio of the bacterial liquid of each bacterial strain is 1:1:1:1:1, 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 thuringiensis is ATCC10792, the Acinetobacter calcoaceticus is ATCC23055, the trichoderma asperellum is TaspHu1, the actinomyces is ATCC27044, and the alcaligenes xylosoxidans is ATCC27061.
2. The composite microbial agent according to claim 1, wherein the total effective viable count of the composite microbial agent is 4.0 x 10 8 -2.0*10 10 cfu/g; the effective viable count in each bacterial liquid is as follows:
3. the composite microbial agent of claim 1, wherein the antibiotic is a tetracycline antibiotic.
4. The composite microbial agent according to claim 1, wherein the pesticide is one or more of a carbamate pesticide, an organochlorine pesticide, and a triazole pesticide.
5. The composite microbial agent according to claim 4, wherein the carbamate pesticide is one or more of carbendazim, carbosulfan, zineb and furadan.
6. The composite microbial agent according to claim 4, wherein the organochlorine pesticide is one or both of endosulfan and trichlorfon.
7. The composite microbial agent according to claim 4, wherein the triazole pesticide is one or both of triazolone and propiconazole.
8. The method for preparing a composite microbial agent according to claim 1, wherein the corresponding strains are cultured to obtain strains with corresponding concentrations, and the strains are mixed to obtain a total effective viable count of 4.0 x 10 8 -2.0*10 10 cfu/g of composite microbial agent.
9. The application method of the composite microbial agent for degrading antibiotics and pesticide residues is characterized in that the composite microbial agent disclosed in any one of claims 1-7 or the composite microbial agent prepared by claim 8 is subjected to soil treatment or crop treatment.
10. The method of use according to claim 9, wherein the soil treatment is: 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: the composite microbial agent is used for spraying the overground part of crops in the growing period of the crops, and the root irrigation treatment can be carried out on underground crops in the growing period.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117535195A (en) * | 2023-11-20 | 2024-02-09 | 宁夏回族自治区食品检测研究院 | Composite microbial agent for degrading antibiotics and pesticide residues and preparation method thereof |
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| WO1996033821A1 (en) * | 1995-04-24 | 1996-10-31 | Shaw Edward A | Remediation of pesticide-contaminated soil |
| CN106434430A (en) * | 2016-09-07 | 2017-02-22 | 中山市润泽生物科技有限公司 | Compound microbial agent for degrading antibiotic and pesticide residues as well as preparation and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO1996033821A1 (en) * | 1995-04-24 | 1996-10-31 | Shaw Edward A | Remediation of pesticide-contaminated soil |
| CN106434430A (en) * | 2016-09-07 | 2017-02-22 | 中山市润泽生物科技有限公司 | Compound microbial agent for degrading antibiotic and pesticide residues as well as preparation and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117535195A (en) * | 2023-11-20 | 2024-02-09 | 宁夏回族自治区食品检测研究院 | Composite microbial agent for degrading antibiotics and pesticide residues and preparation method thereof |
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