CN111057662A - Long-acting quality-guaranteeing compound microbial agent and preparation method thereof - Google Patents
Long-acting quality-guaranteeing compound microbial agent and preparation method thereof Download PDFInfo
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- CN111057662A CN111057662A CN201911227345.XA CN201911227345A CN111057662A CN 111057662 A CN111057662 A CN 111057662A CN 201911227345 A CN201911227345 A CN 201911227345A CN 111057662 A CN111057662 A CN 111057662A
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 59
- 150000001875 compounds Chemical class 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 309
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 103
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 51
- 241000894006 Bacteria Species 0.000 claims abstract description 36
- 230000000243 photosynthetic effect Effects 0.000 claims abstract description 28
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 26
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 26
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 25
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 25
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 24
- 241000187395 Streptomyces microflavus Species 0.000 claims abstract description 24
- 241000186660 Lactobacillus Species 0.000 claims abstract description 22
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 22
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 14
- 241001148471 unidentified anaerobic bacterium Species 0.000 claims description 70
- 239000001963 growth medium Substances 0.000 claims description 65
- 241001148470 aerobic bacillus Species 0.000 claims description 43
- 230000001580 bacterial effect Effects 0.000 claims description 37
- 239000002054 inoculum Substances 0.000 claims description 27
- 239000002131 composite material Substances 0.000 claims description 24
- 240000001046 Lactobacillus acidophilus Species 0.000 claims description 21
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims description 21
- 230000003321 amplification Effects 0.000 claims description 21
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims description 21
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 21
- 239000011573 trace mineral Substances 0.000 claims description 19
- 235000013619 trace mineral Nutrition 0.000 claims description 19
- 230000000844 anti-bacterial effect Effects 0.000 claims description 13
- 239000003899 bactericide agent Substances 0.000 claims description 13
- 241000881860 Paenibacillus mucilaginosus Species 0.000 claims description 12
- 239000007633 bacillus mucilaginosus Substances 0.000 claims description 12
- 229910052796 boron Inorganic materials 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 12
- 229910052742 iron Inorganic materials 0.000 claims description 12
- 229910052725 zinc Inorganic materials 0.000 claims description 12
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 10
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 10
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 9
- 229940081969 saccharomyces cerevisiae Drugs 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 7
- 238000000265 homogenisation Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 229910052802 copper Inorganic materials 0.000 claims description 4
- 229910052748 manganese Inorganic materials 0.000 claims description 4
- 238000000034 method Methods 0.000 claims 3
- 230000005923 long-lasting effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 11
- 239000003337 fertilizer Substances 0.000 abstract description 10
- 244000005700 microbiome Species 0.000 abstract description 7
- 238000009360 aquaculture Methods 0.000 abstract description 4
- 244000144974 aquaculture Species 0.000 abstract description 4
- 238000003860 storage Methods 0.000 abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 31
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 24
- 239000008223 sterile water Substances 0.000 description 24
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 18
- 241000196324 Embryophyta Species 0.000 description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 239000008103 glucose Substances 0.000 description 12
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 12
- 235000019341 magnesium sulphate Nutrition 0.000 description 12
- 239000000126 substance Substances 0.000 description 11
- 239000002689 soil Substances 0.000 description 10
- 239000001888 Peptone Substances 0.000 description 9
- 108010080698 Peptones Proteins 0.000 description 9
- 229910000019 calcium carbonate Inorganic materials 0.000 description 9
- 229940041514 candida albicans extract Drugs 0.000 description 9
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 9
- 235000019319 peptone Nutrition 0.000 description 9
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 9
- 239000012138 yeast extract Substances 0.000 description 9
- 238000009630 liquid culture Methods 0.000 description 8
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 6
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 6
- 235000011130 ammonium sulphate Nutrition 0.000 description 6
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 6
- 235000019797 dipotassium phosphate Nutrition 0.000 description 6
- 239000011777 magnesium Substances 0.000 description 6
- 229940099596 manganese sulfate Drugs 0.000 description 6
- 235000007079 manganese sulphate Nutrition 0.000 description 6
- 239000011702 manganese sulphate Substances 0.000 description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 6
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- 235000017281 sodium acetate Nutrition 0.000 description 6
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- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 5
- 238000013329 compounding Methods 0.000 description 5
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 3
- -1 KH2PO40.5g Substances 0.000 description 3
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 3
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 235000019764 Soybean Meal Nutrition 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- 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 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 235000019270 ammonium chloride Nutrition 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 235000003891 ferrous sulphate Nutrition 0.000 description 3
- 239000011790 ferrous sulphate Substances 0.000 description 3
- 235000013312 flour Nutrition 0.000 description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 3
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 3
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 3
- 239000011572 manganese Substances 0.000 description 3
- 235000013923 monosodium glutamate Nutrition 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 3
- 229920000136 polysorbate Polymers 0.000 description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 229940073490 sodium glutamate Drugs 0.000 description 3
- JXKPEJDQGNYQSM-UHFFFAOYSA-M sodium propionate Chemical compound [Na+].CCC([O-])=O JXKPEJDQGNYQSM-UHFFFAOYSA-M 0.000 description 3
- 239000004324 sodium propionate Substances 0.000 description 3
- 229960003212 sodium propionate Drugs 0.000 description 3
- 235000010334 sodium propionate Nutrition 0.000 description 3
- 239000004455 soybean meal Substances 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 2
- 108010040201 Polymyxins Proteins 0.000 description 2
- 241000235342 Saccharomycetes Species 0.000 description 2
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 2
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 2
- 230000003042 antagnostic effect Effects 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 229910052698 phosphorus Inorganic materials 0.000 description 2
- 239000011574 phosphorus Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 238000011282 treatment Methods 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000001775 anti-pathogenic effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- RKLXDNHNLPUQRB-TVJUEJKUSA-N chembl564271 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]2C(C)SC[C@H](N[C@@H](CC(N)=O)C(=O)NC(=O)[C@@H](NC2=O)CSC1C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)NC(=C)C(=O)N[C@@H](CCCCN)C(O)=O)NC(=O)[C@H]1NC(=O)C(=C\C)/NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]2NC(=O)CNC(=O)[C@@H]3CCCN3C(=O)[C@@H](NC(=O)[C@H]3N[C@@H](CC(C)C)C(=O)NC(=O)C(=C)NC(=O)CC[C@H](NC(=O)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC=4C5=CC=CC=C5NC=4)CSC3)C(O)=O)C(C)SC2)C(C)C)C(C)SC1)C1=CC=CC=C1 RKLXDNHNLPUQRB-TVJUEJKUSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
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- 230000009467 reduction Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 108010082567 subtilin Proteins 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
- C09K17/42—Inorganic compounds mixed with organic active ingredients, e.g. accelerators
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Inorganic Chemistry (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Microbiology (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Soil Sciences (AREA)
- Materials Engineering (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to the technical field of agricultural microorganisms, ecological planting, aquaculture and microbial agents, and particularly relates to a long-acting quality-guaranteeing compound microbial agent and a preparation method thereof. The compound microbial agent comprises a liquid aerobic microbial agent and a liquid anaerobic microbial agent, wherein the liquid aerobic microbial agent comprises: liquid streptomyces microflavus, liquid bacillus subtilis, liquid bacillus amyloliquefaciens and liquid colloidal bacillus; the liquid anaerobic microbial inoculum comprises: liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid acidophilic lactobacillus agent. The compound microorganism liquid microbial inoculum and the preparation method thereof solve the problems of single strain, short storage life and unobvious fertilizer effect of the microorganism microbial inoculum.
Description
Technical Field
The invention relates to the technical field of agricultural microorganisms, ecological planting, aquaculture and microbial agents, and particularly relates to a long-acting quality-guaranteeing compound microbial agent and a preparation method thereof.
Background
At present, the phenomenon of chemical fertilizer abuse in the planting industry and the aquaculture industry of China is very common, and the excessive use of the chemical fertilizer leads to the overproof of harmful substances such as nitrite, ammonia nitrogen, hydrogen sulfide and the like in soil and water, so that the quality of agricultural products is reduced, the health of human bodies is threatened, and the ecological environment is seriously damaged.
In addition, unreasonable application of chemical fertilizers causes continuous reduction of soil conditions, which is not caused by insufficient soil fertility, but excessive nutrient components are consolidated in soil and cannot be absorbed and utilized by crops, and simultaneously, the soil state is greatly changed, so that the survival rate of beneficial microorganisms is greatly reduced. Therefore, it is necessary to add exogenous soil microorganisms. The soil conditioner can release nutrient elements condensed in soil, and can change the types of soil dominant floras through growth promotion and antagonism, so that beneficial floras can obtain the main place, and the soil ecology is recovered.
In recent years, various microbial fertilizers are abundant like bamboo shoots in spring after rain, but products with fertilizer efficiency obviously reaching or exceeding that of the fertilizers are rare, the main reasons are that microbial strains are single and the problems of crop growth cannot be solved, and the microbial inoculum has short storage life and unobvious fertilizer efficiency and is difficult to solve the problems of putrefaction, death of effective bacteria and short storage life of liquid microbial inoculum, namely single microbial inoculum and composite microbial inoculum.
Disclosure of Invention
The invention aims to provide a long-acting quality-guaranteeing composite microbial liquid inoculant and a preparation method thereof, and the composite microbial liquid inoculant and the preparation method thereof solve the problems of single microbial inoculant strain, short storage period and unobvious fertilizer effect.
In order to realize the aim, the invention provides a compound microbial agent with long-acting quality guarantee, the compound microbial liquid agent comprises a liquid aerobic agent and a liquid anaerobic agent,
the liquid aerobic bacterial agent comprises: liquid streptomyces microflavus, liquid bacillus subtilis, liquid bacillus amyloliquefaciens and liquid colloidal bacillus;
the liquid anaerobic microbial inoculum comprises: liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid acidophilic lactobacillus agent.
Further, the mass ratio of the liquid aerobic microbial inoculum to the liquid anaerobic microbial inoculum is 3: 7-7: 3.
Further, the mass parts of the liquid streptomyces microflavus, the liquid bacillus subtilis, the liquid bacillus amyloliquefaciens and the liquid colloidal bacillus are 1-2: 1-3: 1-3: 1 to 3.
Further, the mass parts of the liquid photosynthetic bacteria, the liquid saccharomyces cerevisiae agent, the liquid plant lactobacillus agent and the liquid lactobacillus acidophilus agent are 1-3: 1-3: 1-5: 1 to 5.
Furthermore, the content of aerobic bacteria in each liquid aerobic bacterial agent is more than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic bacterial agent is more than or equal to 2 hundred million/ml.
Furthermore, the microbial agent also comprises trace elements, wherein the trace elements are selected from any one or more of B, Zn, Mn, Cu, Fe and Co.
The invention also provides a preparation method of the compound microbial agent with long-acting quality guarantee, which comprises the following steps:
respectively culturing 4 aerobic bacteria of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens to prepare 4 liquid aerobic bacteria agents;
culturing 4 kinds of anaerobic photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria to prepare a mixed liquid anaerobic microbial agent;
slowly injecting the 4 liquid aerobic bactericides into the composite liquid anaerobic bactericides, simultaneously adding trace elements, uniformly stirring, standing, and obtaining the composite microbial bactericides after homogenization is stable.
Further, the mass parts of the liquid streptomyces microflavus, the liquid bacillus subtilis, the liquid bacillus amyloliquefaciens and the liquid colloidal bacillus are 1-2: 1-3: 1-3: 1 to 3.
Further, the step of preparing the liquid aerobic bacterial agent comprises the following steps:
respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 100-140 r/min and pH value of 6.0-7.5 to obtain 5 primary aerobic bacteria liquids;
and then respectively taking 4 primary aerobic bacterial liquids, inoculating corresponding strains into the 4 liquid aerobic bacterial culture media, and carrying out secondary amplification culture on the strains at the temperature of 33-37 ℃ and the speed of 100-140 r/min to obtain 4 secondary aerobic bacterial liquids, wherein the inoculation amount of each primary aerobic bacterial liquid is 0.7-1.3% of the mass of the liquid aerobic bacterial culture medium.
Further, the step of preparing the liquid anaerobic microbial inoculum comprises the following steps:
respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains to the 4 liquid anaerobic bacteria culture media, standing at the temperature of 30-37 ℃, and performing primary amplification culture of the strains to obtain 3 primary anaerobic bacteria liquids;
respectively inoculating 4 kinds of first-stage anaerobic bacteria liquid into 4 kinds of liquid anaerobic bacteria culture media to obtain corresponding strains, standing at the temperature of 30-37 ℃, and performing second-stage amplification culture on the strains to obtain 4 kinds of second-stage anaerobic bacteria liquid, wherein the inoculation amount of each first-stage anaerobic bacteria liquid is 0.7-1.3% of the mass of the liquid anaerobic bacteria culture medium;
according to the following steps, 1-3 parts by mass of liquid photosynthetic bacteria, liquid saccharomyces cerevisiae microbial inoculum, liquid plant lactobacillus microbial inoculum and liquid lactobacillus acidophilus microbial inoculum: 1-3: 1-5: and (3) directly and uniformly mixing 4 secondary anaerobic bacteria liquids in a proportion of 1-5 to obtain the composite liquid anaerobic microbial inoculum.
Advantageous effects
The beneficial microbe is added into soil and culture water to decompose pollutant, purify water, increase dissolved oxygen and reduce fertilizer consumption greatly.
The photosynthetic bacteria belong to independent nutrient microorganisms, can convert absorbed substances into bioactive substances such as saccharides, amino acids, vitamins and the like, and the synthesized substances can provide available substance nutrition for lactobacillus, saccharomycetes and bacillus and promote the growth of the lactobacillus, the saccharomycetes and the bacillus. Meanwhile, the independent culture of each flora ensures the normal propagation and development of the flora, so that the flora can reach the optimal growth node and then is compounded. After the compounding is finished, the flora is influenced by certain condition difference, the growth and the propagation of the single flora are slowed down, the non-antagonistic symbiotic environmental conditions are formed, and the preservation time under the normal temperature condition is greatly prolonged. Meanwhile, the compounding of the aerobic bacteria and the anaerobic bacteria can meet the requirement that at least one kind of bacteria can play the normal proliferation and secretion functions under certain extreme conditions, so that the normal effect is ensured.
The number of effective viable bacteria is large, the content of a single flora is not lower than 2 hundred million/ml, each flora can play different functions, and the effective viable bacteria and the single flora are mutually coordinated and proliferated, so that the functions are comprehensive;
the growth conditions of each flora are different, and the excellent symbiosis is realized under the condition that the floras are not antagonistic to each other. However, because the growth conditions are not optimal, the growth of each flora is inhibited to a certain extent, so that the development speed is slowed down, multiple bacteria are coordinated and symbiotic with each other, and the shelf life of the bacteria is prolonged from half a year to one year;
the proliferation and growth of the flora need a proper environment, and the compounding of the aerobic bacteria and the anaerobic bacteria can ensure that one or all of the aerobic bacteria and the anaerobic bacteria can normally play a role under the aerobic condition, the anaerobic condition and the facultative anaerobic condition, thereby ensuring the efficacy of the bacteria. Under aerobic conditions, the liquid bacillus subtilis can be normally propagated and developed and secrete active substances such as subtilin, polymyxin and the like to inhibit pathogenic bacteria. Under anaerobic or dark conditions, photosynthetic bacteria degrade toxic substances such as nitrite or sulfide.
Detailed Description
The following describes in detail embodiments of the present invention.
The composite microbial liquid inoculum comprises 4 liquid aerobic inocula cultured by a liquid culture medium and 4 liquid anaerobic inoculas cultured by the liquid culture medium, and the mass ratio of the liquid aerobic inoculas to the liquid anaerobic inoculas is 3: 7-7: 3.
The liquid aerobic bacterial agent comprises: liquid streptomyces microflavus agent, liquid bacillus subtilis agent, liquid bacillus amyloliquefaciens agent and liquid colloidal bacillus agent.
The liquid anaerobic microbial inoculum comprises: liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid acidophilic lactobacillus agent.
The liquid aerobic bacterial agent comprises: 1-2 parts by mass of a liquid streptomyces microflavus microbial inoculum; 1-3 parts of a liquid bacillus subtilis preparation; 1-3 parts of liquid bacillus amyloliquefaciens and 1-3 parts of liquid colloidal bacillus agent.
The liquid anaerobic microbial inoculum comprises: 1-3 parts by mass of liquid photosynthetic bacteria; 1-3 parts by mass of a liquid saccharomyces cerevisiae microbial inoculum; 1-5 parts by mass of a liquid plant lactobacillus agent; 1-5 parts of liquid lactobacillus acidophilus.
Wherein, the content of aerobic bacteria in each liquid aerobic bacteria agent is more than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic bacteria agent is more than or equal to 2 hundred million/ml.
The microbial agent also comprises trace elements, wherein the trace elements are selected from any one or more of B, Zn, Mn, Cu, Fe and Co.
A preparation method of a compound microbial agent with long-acting quality guarantee comprises the following steps:
1. respectively culturing 4 aerobic bacteria of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens to prepare 4 liquid aerobic bacteria agents,
(1) respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 100-140 r/min and pH value of 6.0-7.5 to obtain 5 primary aerobic bacteria liquids;
(2) respectively inoculating 4 kinds of first-stage aerobic bacterial liquid into 4 kinds of liquid aerobic bacterial culture media to obtain corresponding strains, and performing second-stage amplification culture on the strains at the temperature of 33-37 ℃ and the speed of 100-140 r/min to obtain 4 kinds of second-stage aerobic bacterial liquids, wherein the inoculation amount of each first-stage aerobic bacterial liquid is 0.7-1.3% of the quality of the liquid aerobic bacterial culture medium;
2. culturing 4 kinds of anaerobic photosynthetic bacteria, Saccharomyces cerevisiae, Lactobacillus plantarum and Lactobacillus acidophilus to obtain mixed liquid anaerobic bacteria,
(1) respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains to the 4 liquid anaerobic bacteria culture media, standing at the temperature of 30-37 ℃, and performing primary amplification culture on the strains to obtain 3 primary anaerobic bacteria liquids;
(2) respectively inoculating 4 kinds of first-stage anaerobic bacteria liquid into 4 kinds of liquid anaerobic bacteria culture media to obtain corresponding strains, standing at the temperature of 30-37 ℃, and performing second-stage amplification culture on the strains to obtain 4 kinds of second-stage anaerobic bacteria liquid, wherein the inoculation amount of each first-stage anaerobic bacteria liquid is 0.7-1.3% of the mass of the liquid anaerobic bacteria culture medium;
(3) according to the following steps, 1-3 parts by mass of liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid lactobacillus acidophilus agent: 1-3: 1-5: 1-5, directly and uniformly mixing 4 secondary anaerobic bacteria liquids to obtain a composite liquid anaerobic microbial inoculum;
3. and (2) mixing 4 liquid aerobic bactericides according to the mass parts of 1-2: 1-3: 1-3: slowly injecting the mixture into the composite liquid anaerobic microbial inoculum according to the proportion of 1-3, and simultaneously adding trace elements, wherein the trace elements are selected from any one or more of B, Zn, Mn, Cu, Fe and Co. Stirring uniformly, standing, and obtaining the composite microbial inoculum after homogenization and stabilization.
The components of the culture medium:
1.4 aerobic liquid microbial inoculum culture medium
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
B, bacillus amyloliquefaciens: 5g of sodium glutamate, 15g of glucose, KH2PO40.5g, Mg SO40.2g, MnSO42.0 Mg and 1L of sterile water.
B, Bacillus mucilaginosus: 24g of glucose, 1g of ammonium sulfate, 9g of bean cake, 2g of calcium carbonate, 2g of magnesium sulfate, 4g of monopotassium phosphate, 0.1g of ferric chloride and 1L of sterile water.
2.4 anaerobic liquid microbial inoculum culture medium
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Brewing yeast: 60g of peptone, 30g of maltose, 40.5g of MgSO40, 40.01g of ZnSO40, 1g of KCl and 1L of sterile water.
Plant lactobacillus: 30g of sucrose, 50g of yeast extract, 5g of sodium acetate, 2g of dipotassium phosphate, 2g of diammonium hydrogen citrate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 2g of tween-801 m L, 2g of calcium carbonate and 1L of sterile water.
Lactobacillus acidophilus: 61g of glucose, 40g of peptone, 43.56g of Na2HPO43, 0.44g of citric acid, 6g of MgS 043 g potato extract, 801 g of Tween and 1L of sterile water.
Example 1
The compound microbial agent comprises 4 liquid aerobic microbial agents cultured by a liquid culture medium and 4 liquid anaerobic microbial agents cultured by the liquid culture medium, wherein the mass ratio of the liquid aerobic microbial agents to the liquid anaerobic microbial agents is 2: 3.
The liquid aerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
2 parts of liquid streptomyces microflavus microbial inoculum; 3 parts of liquid bacillus subtilis preparation; 2 parts of liquid bacillus amyloliquefaciens and 3 parts of liquid colloidal bacillus agent.
The liquid anaerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
3 parts of liquid photosynthetic bacteria; 2 parts of liquid saccharomyces cerevisiae microbial inoculum; 5 parts of liquid plant lactobacillus; 5 parts of liquid lactobacillus acidophilus.
Wherein, the content of aerobic bacteria in each liquid aerobic bacteria agent is more than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic bacteria agent is more than or equal to 2 hundred million/ml. The microbial liquid inoculum also comprises trace elements, wherein the trace elements are four or 3 of B, Zn, Fe and Co.
The preparation method comprises the following steps:
1. respectively culturing 4 aerobic bacteria to obtain 4 liquid aerobic bacteria agents,
(1) respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH value of 6.5-7.0 to obtain 4 primary aerobic bacteria liquids;
(2) then respectively taking 4 primary aerobic bacterial liquids, inoculating corresponding strains into 4 liquid aerobic bacterial culture media, and carrying out secondary expansion culture on the strains at 33-37 ℃ and 120r/min to obtain 4 secondary aerobic bacterial liquids, wherein the inoculation amount of each primary aerobic bacterial liquid is 1.0 percent of the mass of the liquid aerobic bacterial culture medium;
2.4 kinds of anaerobes are cultured to prepare a mixed liquid anaerobe agent
(1) Respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains into the 4 liquid anaerobic bacteria culture media, standing at the temperature of 33-37 ℃, and performing primary amplification culture on the strains to obtain 3 primary anaerobic bacteria liquid;
(2) then 4 kinds of first-stage anaerobic bacteria liquid are respectively taken to inoculate corresponding strains in 4 kinds of liquid anaerobic bacteria culture media,
standing at 33-37 ℃, and performing secondary amplification culture on strains to obtain 4 secondary anaerobic bacteria liquids, wherein the inoculation amount of each primary anaerobic bacteria liquid is 1.0% of the mass of the liquid anaerobic bacteria culture medium;
(3) directly and uniformly mixing the 4 kinds of second-stage anaerobic bacteria liquid according to the formula proportion of the technical scheme to obtain a composite liquid anaerobic microbial inoculum;
3. slowly injecting the 4 liquid aerobic bactericides into the composite liquid anaerobic bactericides according to the proportion, and simultaneously adding trace elements, wherein the trace elements are selected from any one or a mixture of at least two of B, Zn, Fe and Co. Stirring uniformly, standing, and obtaining the compound microbial agent after homogenization is stable.
The components of the culture medium:
1.4 aerobic liquid microbial inoculum culture medium
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
B, bacillus amyloliquefaciens: 5g of sodium glutamate, 15g of glucose, KH2PO40.5g, Mg SO40.2g, MnSO42.0 Mg and 1L of sterile water.
B, Bacillus mucilaginosus: 24g of glucose, 1g of ammonium sulfate, 9g of bean cake, 2g of calcium carbonate, 2g of magnesium sulfate, 4g of monopotassium phosphate, 0.1g of ferric chloride and 1L of sterile water.
2.4 anaerobic liquid microbial inoculum culture medium
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Brewing yeast: 60g of peptone, 30g of maltose, 40.5g of MgSO40, 40.01g of ZnSO40, 1g of KCl and 1L of sterile water.
Plant lactobacillus: 30g of sucrose, 50g of yeast extract, 5g of sodium acetate, 2g of dipotassium phosphate, 2g of diammonium hydrogen citrate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 2g of tween-801 m L, 2g of calcium carbonate and 1L of sterile water.
Lactobacillus acidophilus: 61g of glucose, 40g of peptone, 43.56g of Na2HPO43, 0.44g of citric acid, 6g of MgS 043 g potato extract, 801 g of Tween and 1L of sterile water.
Example 2
The compound microbial agent comprises 4 liquid aerobic microbial agents cultured by a liquid culture medium and 4 liquid anaerobic microbial agents cultured by the liquid culture medium, wherein the mass ratio of the liquid aerobic microbial agents to the liquid anaerobic microbial agents is 10: 11.
The liquid aerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
1 part by mass of a liquid streptomyces microflavus microbial inoculum; 3 parts of liquid bacillus subtilis preparation; 3 parts of liquid bacillus amyloliquefaciens and 3 parts of liquid colloidal bacillus agent.
The liquid anaerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
2 parts of liquid photosynthetic bacteria; 3 parts of a liquid saccharomyces cerevisiae microbial inoculum; 3 parts of liquid plant lactobacillus; 3 parts of liquid lactobacillus acidophilus.
Wherein, the content of aerobic bacteria in each liquid aerobic bacteria agent is more than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic bacteria agent is more than or equal to 2 hundred million/ml. The microbial liquid inoculum also comprises trace elements which are B, Zn, Fe and Co 4 or a mixture of 3 of the B, Zn, Fe and Co.
The preparation method comprises the following steps:
1. respectively culturing 4 aerobic bacteria to obtain 4 liquid aerobic bacteria agents,
(1) respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH value of 6.5-7.0 to obtain 4 primary aerobic bacteria liquids;
(2) then respectively taking 4 primary aerobic bacterial liquids, inoculating corresponding strains into 4 liquid aerobic bacterial culture media, and carrying out secondary expansion culture on the strains at 33-37 ℃ and 120r/min to obtain 4 secondary aerobic bacterial liquids, wherein the inoculation amount of each primary aerobic bacterial liquid is 1.0 percent of the mass of the liquid aerobic bacterial culture medium;
2.4 kinds of anaerobes are cultured to prepare a mixed liquid anaerobe agent
(1) Respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains into the 4 liquid anaerobic bacteria culture media, standing at the temperature of 33-37 ℃, and performing primary amplification culture on the strains to obtain 4 primary anaerobic bacteria liquid;
(2) then 4 kinds of first-stage anaerobic bacteria liquid are respectively taken to inoculate corresponding strains in 4 kinds of liquid anaerobic bacteria culture media,
standing at 33-37 ℃, and performing secondary amplification culture on strains to obtain 4 secondary anaerobic bacteria liquids, wherein the inoculation amount of each primary anaerobic bacteria liquid is 1.0% of the mass of the liquid anaerobic bacteria culture medium;
(3) directly and uniformly mixing the 4 kinds of second-stage anaerobic bacteria liquid according to the formula proportion of the technical scheme to obtain a composite liquid anaerobic microbial inoculum;
3. slowly injecting the 4 liquid aerobic bactericides into the composite liquid anaerobic bactericides according to the proportion, and simultaneously adding trace elements, wherein the trace elements are selected from any one or a mixture of at least two of B, Zn, Fe and Co. Stirring uniformly, standing, and obtaining the compound microbial agent after homogenization is stable.
Example 3
The compound microbial agent comprises 5 liquid aerobic microbial agents cultured by a liquid culture medium and 3 liquid anaerobic microbial agents cultured by the liquid culture medium, wherein the mass ratio of the liquid aerobic microbial agents to the liquid anaerobic microbial agents is 7: 10.
The liquid aerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
1 part by mass of a liquid streptomyces microflavus microbial inoculum; 2 parts of liquid bacillus subtilis preparation by mass; 2 parts of liquid bacillus amyloliquefaciens and 2 parts of liquid colloidal bacillus agent.
The liquid anaerobic microbial inoculum consists of the following microbial inoculum in parts by mass:
2 parts of liquid photosynthetic bacteria; 2 parts of liquid saccharomyces cerevisiae microbial inoculum; 3 parts of liquid plant lactobacillus; 3 parts of liquid lactobacillus acidophilus.
Wherein, the content of aerobic bacteria in each liquid aerobic bacteria agent is more than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic bacteria agent is more than or equal to 2 hundred million/ml. The microbial liquid inoculum also comprises trace elements which are B, Zn, Fe and Co 4 or a mixture of 3 of the B, Zn, Fe and Co.
The preparation method comprises the following steps:
1. respectively culturing 4 aerobic bacteria to obtain 4 liquid aerobic bacteria agents,
(1) respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 120r/min and pH value of 6.5-7.0 to obtain 4 primary aerobic bacteria liquids;
(2) then respectively taking 4 primary aerobic bacterial liquids, inoculating corresponding strains into 4 liquid aerobic bacterial culture media, and carrying out secondary expansion culture on the strains at 33-37 ℃ and 120r/min to obtain 4 secondary aerobic bacterial liquids, wherein the inoculation amount of each primary aerobic bacterial liquid is 1.0 percent of the mass of the liquid aerobic bacterial culture medium;
2.4 kinds of anaerobes are cultured to prepare a mixed liquid anaerobe agent
(1) Respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains into the 4 liquid anaerobic bacteria culture media, standing at the temperature of 33-37 ℃, and performing primary amplification culture on the strains to obtain 4 primary anaerobic bacteria liquid;
(2) then 4 kinds of first-stage anaerobic bacteria liquid are respectively taken to inoculate corresponding strains in 4 kinds of liquid anaerobic bacteria culture media,
standing at 33-37 ℃, and performing secondary amplification culture on strains to obtain 4 secondary anaerobic bacteria liquids, wherein the inoculation amount of each primary anaerobic bacteria liquid is 1.0% of the mass of the liquid anaerobic bacteria culture medium;
(3) directly and uniformly mixing the 4 kinds of second-stage anaerobic bacteria liquid according to the formula proportion of the technical scheme to obtain a composite liquid anaerobic microbial inoculum;
3. slowly injecting the 4 liquid aerobic bactericides into the composite liquid anaerobic bactericides according to the proportion, and simultaneously adding trace elements, wherein the trace elements are selected from any one or a mixture of at least two of B, Zn, Fe and Co. Stirring uniformly, standing, and obtaining the compound microbial agent after homogenization is stable.
The components of the culture medium:
1.4 aerobic liquid microbial inoculum culture medium
Streptomyces microflavus: KNO 31 g, 20g of soluble starch, 40.5g of K2HPO40, 40.5g of MgSO40, 0.5g of NaCl, 40.01g of FeSO40 and 1L of sterile water.
B, bacillus subtilis: 40g of soybean meal, 20g of corn flour, 15g of glucose, 3g of dipotassium phosphate, 1.5g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate, 0.35g of ammonium sulfate, 0.2g of yeast extract powder, 0.2g of manganese sulfate, 0.1g of ferrous sulfate, 0.1g of calcium carbonate and 1L of sterile water.
B, bacillus amyloliquefaciens: 5g of sodium glutamate, 15g of glucose, KH2PO40.5g, Mg SO40.2g, MnSO42.0 Mg and 1L of sterile water.
B, Bacillus mucilaginosus: 24g of glucose, 1g of ammonium sulfate, 9g of bean cake, 2g of calcium carbonate, 2g of magnesium sulfate, 4g of monopotassium phosphate, 0.1g of ferric chloride and 1L of sterile water.
2.4 anaerobic liquid microbial inoculum culture medium
Photosynthetic bacteria: 1g of ammonium chloride, 1g of sodium bicarbonate, 1g of sodium acetate, 1g of sodium chloride, 0.2g of dipotassium hydrogen phosphate, 0.2g of sodium propionate, 0.2g of magnesium sulfate, 0.2g of peptone, 0.1g of yeast extract and 1L of sterile water.
Brewing yeast: 60g of peptone, 30g of maltose, 40.5g of MgSO40, 40.01g of ZnSO40, 1g of KCl and 1L of sterile water.
Plant lactobacillus: 30g of sucrose, 50g of yeast extract, 5g of sodium acetate, 2g of dipotassium phosphate, 2g of diammonium hydrogen citrate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 2g of tween-801 m L, 2g of calcium carbonate and 1L of sterile water.
Lactobacillus acidophilus: 61g of glucose, 40g of peptone, 43.56g of Na2HPO43, 0.44g of citric acid, 6g of MgS 043 g potato extract, 801 g of Tween and 1L of sterile water.
Example 4
Application of microbial agent in aquaculture
And (3) test treatment: set 2 parallel test groups, 3 treatments per group, control: no product is used; t1: 1.5ppm for the first 3 times and 1.0ppm for the second 2 times; t2: the preparation is administered every 10 days at 3.0ppm for the first 3 times and 2.0ppm for the second 2 times.
Measurement indexes are as follows: and measuring indexes such as water ammonia nitrogen, nitrite, chemical oxygen demand, total phosphorus and the like every 7 days.
TABLE 1 amount of inoculum
TABLE 2 Effect on Ammonia Nitrogen (mg/L) in ponds
TABLE 3 Effect on nitrite in ponds
TABLE 4 Effect on Pond COD
TABLE 5 Effect on Total phosphorus in ponds
In the invention, different floras are compounded to have multiple functions, such as bacillus subtilis secreting anti-pathogenic bacteria such as polymyxin and the like, and inhibiting soil-borne diseases caused by harmful floras; the photosynthetic bacteria can degrade harmful substances such as nitrite, sulfide and the like in the water body and purify the water quality; the lactobacillus plantarum can produce acid in large quantity, so that the pH value in water is stable and not increased, and the produced acidic substances can degrade heavy metals and the like. Through reasonable proportioning and compounding, the multifunctional effect is exerted while the antagonism among different floras is avoided, and the compounding effect of multiple strains is shown in table 6.
TABLE 6 comparison of the Using effects of the culture Water
Claims (10)
1. A composite microbial agent with long-acting quality guarantee is disclosed, the composite microbial liquid agent comprises a liquid aerobic agent and a liquid anaerobic agent,
the method is characterized in that the liquid aerobic bacterial agent comprises the following components: liquid streptomyces microflavus, liquid bacillus subtilis, liquid bacillus amyloliquefaciens and liquid colloidal bacillus;
the liquid anaerobic microbial inoculum comprises: liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid acidophilic lactobacillus agent.
2. The long-acting quality-guaranteeing compound microbial inoculant according to claim 1, wherein the mass ratio of the liquid aerobic inoculant to the liquid anaerobic inoculant is 3: 7-7: 3.
3. The long-acting quality-guaranteeing compound microbial inoculant according to claim 1, wherein the mass parts of the liquid streptomyces microflavus inoculant, the liquid bacillus subtilis inoculant, the liquid bacillus amyloliquefaciens inoculant and the liquid colloidal bacillus inoculant are (1-2): 1-3: 1-3: 1 to 3.
4. The long-acting quality-guaranteeing compound microbial inoculant according to claim 1, wherein the mass parts of the liquid photosynthetic bacteria, the liquid saccharomyces cerevisiae inoculant, the liquid plant lactobacillus inoculant and the liquid lactobacillus acidophilus inoculant are 1-3: 1-3: 1-5: 1 to 5.
5. The long-acting quality-guaranteeing compound microbial inoculant according to claim 1, wherein the content of aerobic bacteria in each liquid aerobic inoculant is greater than or equal to 2 hundred million/ml, and the content of anaerobic bacteria in each liquid anaerobic inoculant is greater than or equal to 2 hundred million/ml.
6. The long-acting quality-guaranteeing composite microbial agent according to claim 1, further comprising trace elements selected from any one or more of B, Zn, Mn, Cu, Fe and Co.
7. A preparation method of a compound microbial agent with long-acting quality guarantee is characterized by comprising the following steps:
respectively culturing 4 aerobic bacteria of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens to prepare 4 liquid aerobic bacteria agents;
culturing 4 kinds of anaerobic photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria to prepare a mixed liquid anaerobic microbial agent;
slowly injecting the 4 liquid aerobic bactericides into the composite liquid anaerobic bactericides, simultaneously adding the trace elements, uniformly stirring, standing, and obtaining the composite microbial bactericides after homogenization is stable.
8. The preparation method of the compound microbial agent with long-acting quality guarantee according to claim 7, wherein the mass parts of the liquid streptomyces microflavus agent, the liquid bacillus subtilis agent, the liquid bacillus amyloliquefaciens agent and the liquid colloidal bacillus agent are 1-2: 1-3: 1-3: 1 to 3.
9. The method for preparing a long-acting quality-guaranteeing composite microbial inoculant according to claim 7, wherein the step of preparing the liquid aerobic inoculant comprises the following steps:
respectively preparing liquid aerobic bacteria culture media of bacillus mucilaginosus, streptomyces microflavus, bacillus subtilis and bacillus amyloliquefaciens, respectively inoculating corresponding strains into the 4 liquid aerobic bacteria culture media, and performing primary amplification culture on the strains under the conditions of 33-37 ℃, 100-140 r/min and pH value of 6.0-7.5 to obtain 5 primary aerobic bacteria liquids;
and then respectively taking 4 primary aerobic bacterial liquids, inoculating corresponding strains into the 4 liquid aerobic bacterial culture media, and carrying out secondary amplification culture on the strains at the temperature of 33-37 ℃ and the speed of 100-140 r/min to obtain 4 secondary aerobic bacterial liquids, wherein the inoculation amount of each primary aerobic bacterial liquid is 0.7-1.3% of the mass of the liquid aerobic bacterial culture medium.
10. The method for preparing a long-lasting quality-guaranteeing composite microbial inoculant according to claim 7, wherein the step of preparing the liquid anaerobic inoculant comprises the steps of:
respectively preparing photosynthetic bacteria, saccharomyces cerevisiae, lactobacillus plantarum and lactobacillus acidophilus liquid anaerobic bacteria culture media, respectively inoculating corresponding strains into the 4 liquid anaerobic bacteria culture media, standing at the temperature of 30-37 ℃, and performing primary amplification culture on the strains to obtain 3 primary anaerobic bacteria liquid;
respectively inoculating 4 kinds of first-stage anaerobic bacteria liquid into 4 kinds of liquid anaerobic bacteria culture media to obtain corresponding strains, standing at the temperature of 30-37 ℃, and performing second-stage amplification culture on the strains to obtain 4 kinds of second-stage anaerobic bacteria liquid, wherein the inoculation amount of each first-stage anaerobic bacteria liquid is 0.7-1.3% of the mass of the liquid anaerobic bacteria culture medium;
according to the following steps, 1-3 parts by mass of liquid photosynthetic bacteria, liquid saccharomyces cerevisiae agent, liquid plant lactobacillus agent and liquid lactobacillus acidophilus agent: 1-3: 1-5: and (3) directly and uniformly mixing 4 secondary anaerobic bacteria liquids in a proportion of 1-5 to obtain the composite liquid anaerobic microbial inoculum.
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|---|---|---|---|---|
| CN113249262A (en) * | 2021-05-31 | 2021-08-13 | 中诚国联（河南）生物科技有限公司 | Compound microbial agent for preventing and treating diseases and application thereof |
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| CN111057662B (en) | 2022-01-28 |
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