CN112812990A - Compound microbial agent for excrement treatment and application and compounding method thereof - Google Patents
Compound microbial agent for excrement treatment and application and compounding method thereof Download PDFInfo
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 29
- 150000001875 compounds Chemical class 0.000 title claims abstract description 25
- 238000013329 compounding Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 14
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 61
- 241000186361 Actinobacteria <class> Species 0.000 claims abstract description 30
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 20
- 239000001963 growth medium Substances 0.000 claims description 61
- 238000011218 seed culture Methods 0.000 claims description 50
- 238000010563 solid-state fermentation Methods 0.000 claims description 46
- 238000000855 fermentation Methods 0.000 claims description 45
- 230000004151 fermentation Effects 0.000 claims description 45
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 30
- 239000001888 Peptone Substances 0.000 claims description 30
- 108010080698 Peptones Proteins 0.000 claims description 30
- 235000019319 peptone Nutrition 0.000 claims description 30
- 238000012258 culturing Methods 0.000 claims description 25
- 239000000758 substrate Substances 0.000 claims description 25
- 239000007787 solid Substances 0.000 claims description 24
- 241000186046 Actinomyces Species 0.000 claims description 23
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 21
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 21
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 21
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 20
- 238000012807 shake-flask culturing Methods 0.000 claims description 20
- 239000000725 suspension Substances 0.000 claims description 20
- 241000894006 Bacteria Species 0.000 claims description 15
- 235000015278 beef Nutrition 0.000 claims description 15
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims description 15
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims description 15
- 235000019838 diammonium phosphate Nutrition 0.000 claims description 15
- 238000001035 drying Methods 0.000 claims description 15
- 239000007788 liquid Substances 0.000 claims description 15
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 15
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 15
- 235000015097 nutrients Nutrition 0.000 claims description 15
- 238000004321 preservation Methods 0.000 claims description 12
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 11
- 238000009629 microbiological culture Methods 0.000 claims description 11
- 241001446247 uncultured actinomycete Species 0.000 claims description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- LLSDKQJKOVVTOJ-UHFFFAOYSA-L calcium chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Ca+2] LLSDKQJKOVVTOJ-UHFFFAOYSA-L 0.000 claims description 10
- 235000013312 flour Nutrition 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000011081 inoculation Methods 0.000 claims description 10
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 10
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 10
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 10
- 239000000463 material Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 235000010333 potassium nitrate Nutrition 0.000 claims description 10
- 235000013619 trace mineral Nutrition 0.000 claims description 10
- 239000011573 trace mineral Substances 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 9
- 239000000440 bentonite Substances 0.000 claims description 7
- 229910000278 bentonite Inorganic materials 0.000 claims description 7
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 claims description 7
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- 229910000357 manganese(II) sulfate Inorganic materials 0.000 claims description 6
- 229920001817 Agar Polymers 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 5
- 244000068988 Glycine max Species 0.000 claims description 5
- 235000010469 Glycine max Nutrition 0.000 claims description 5
- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 241000203775 Thermoactinomyces Species 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- 239000008272 agar Substances 0.000 claims description 5
- 239000001110 calcium chloride Substances 0.000 claims description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 5
- 235000011148 calcium chloride Nutrition 0.000 claims description 5
- 229910052799 carbon Inorganic materials 0.000 claims description 5
- 239000011248 coating agent Substances 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 235000015099 wheat brans Nutrition 0.000 claims description 5
- 239000002054 inoculum Substances 0.000 claims 6
- 239000002131 composite material Substances 0.000 claims 2
- 230000002550 fecal effect Effects 0.000 claims 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 244000000010 microbial pathogen Species 0.000 abstract description 5
- 239000003895 organic fertilizer Substances 0.000 abstract description 5
- 241000209094 Oryza Species 0.000 description 4
- 241000203780 Thermobifida fusca Species 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000004519 manufacturing process Methods 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
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/80—Separation, elimination or disposal of harmful substances during the treatment
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Life Sciences & Earth Sciences (AREA)
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- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Zoology (AREA)
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- Toxicology (AREA)
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- Treatment Of Sludge (AREA)
Abstract
The invention discloses a compound microbial agent for excrement treatment, which comprises thermophilic actinomycetes and bacillus licheniformis. The invention is applied to the biological treatment of excrement. Meanwhile, the invention also discloses a compounding method of the compound microbial agent for excrement treatment. According to the invention, the dry toilet excrement which is difficult to utilize and has heavy peculiar smell is fermented into a treated product with the pathogenic microorganism excrement coliform flora content meeting the harmless requirement of the organic fertilizer, the peculiar smell is reduced, and the total amount is reduced.
Description
Technical Field
The invention relates to the field of excrement treatment, in particular to a compound microbial agent for excrement treatment and an application and compounding method thereof.
Background
Many parts of the world lack water sources, and drinking water necessary for life is difficult and becomes a luxury when used as toilet water; meanwhile, as the natural degradation period of excrement is long, the survival time and the survival quantity of harmful bacteria are increased, and particularly in alpine regions, the environment is harmed if the harmful bacteria are not treated correspondingly. The excrement is fermented by the compound microbial agent, so that water for a toilet can be saved, water resources are saved, and the fermented excrement of the dry toilet is effectively controlled in pathogenic microorganism content and can be better converted into organic fertilizer required by agriculture.
Disclosure of Invention
The invention aims to provide a compound microbial agent for excrement treatment.
The invention also provides a compound microbial agent for excrement treatment, which is applied to the biological treatment of excrement.
The invention also provides a compounding method of the compound microbial agent for excrement treatment.
The invention has the innovation points that the dry toilet excrement which is difficult to utilize and has heavy peculiar smell is fermented into a treated product with the pathogenic microorganism excrement coliform flora content meeting the harmless requirement of the organic fertilizer, the peculiar smell is reduced and the total amount is reduced, meanwhile, the water in the excrement can be evaporated under the heat production effect of the fermentation process, the electric energy is saved, the water resource does not need to be reused, and the purpose of saving and utilizing the resources is achieved.
In order to achieve the purpose, the technical scheme of the invention is as follows: a compound microbial agent for treating excrement comprises thermophilic actinomycetes Thermobifida fusca and Bacillus licheniformis, wherein the thermophilic actinomycetes Thermobifida fusca is preserved in China general microbiological culture Collection center (China general microbiological culture Collection management Committee) at 11 months and 09 days 2020, the preservation address is No. 3 of West Lu No. 1 of the sunward area of Beijing, and the preservation number is CGMCC No. 21143; bacillus licheniformis is preserved in China general microbiological culture Collection center on 09.11.2020, with the preservation address of No. 3 Siro-1 of Beijing republic of south china, and the preservation number of CGMCC No. 21144; the compounding ratio of the thermophilic actinomycete Thermobifida fusca to the Bacillus licheniformis is 1: 5-7. According to the invention, the dry toilet excrement which is difficult to utilize and has heavy peculiar smell is fermented into a treated product with the pathogenic microorganism excrement coliform flora content meeting the harmless requirement of the organic fertilizer, the peculiar smell is reduced, and the total amount is reduced.
Furthermore, the number of effective viable bacteria of the Thermoactinomyces is more than or equal to 2 multiplied by 108cfu/g, the effective viable count of the bacillus licheniformis is more than or equal to 2 multiplied by 108 cfu/g。
A compound microbial preparation for treating excrement is used for biological treatment of excrement.
A compound method of a compound microbial agent for excrement treatment comprises the following steps:
(1) solid state fermentation: respectively carrying out solid state fermentation on the thermophilic actinomyces and the bacillus licheniformis;
(2) and (3) drying: drying the solid fermented thermophilic actinomycetes and bacillus licheniformis at the temperature of 70 ℃, and obtaining thermophilic actinomycetes products and bacillus licheniformis products with the water content of less than 10 percent after drying;
(3) mixing and crushing: mixing the thermophilic actinomyces product and the bacillus licheniformis product according to the proportion of 1: 5-7 to form a mixed product, crushing the mixed product, and adding an auxiliary material according to the proportion of the bacteria content meeting the requirement to obtain a finished product, wherein the auxiliary material is 150-400 meshes of bentonite.
Further, the compound microbial agent for treating the excrement has the bacterium content of 1.0 multiplied by 10 in the fresh substrate of the thermophilic actinomycete product10cfu/g, the bacteria content in the fresh substrate of the fermentation product of the bacillus licheniformis is 2.5 multiplied by 1010 cfu/g。
Further, the bentonite is 300 mesh.
Further, the solid state fermentation of the actinomyces thermophilus adopts the following steps:
(1) and (3) shake flask culture: inoculating the thermophilic actinomycete strain into a shake flask culture medium from a preserved inclined plane, standing, and performing activated culture for 12-14 h at the temperature of 55-60 ℃; the formula of the shake flask culture medium comprises peptone, glucose and water, wherein the mass ratio of the peptone to the glucose to the water is 25:15:1000, and the pH value is 7.0-7.5;
(2) seed culture: inoculating the thermophilic actinomyces in the shake flask culture medium into a seed culture medium according to the inoculation amount of 2.0-3.0%, and culturing at the rotation speed of 30-40 r/min and the temperature of 55-60 ℃ for 10-12 h to obtain a seed solution; counting the thermophilic actinomycetes in the seed liquid, wherein the formula of the seed culture medium comprises peptone, beef extract and water, the mass ratio of the peptone to the beef extract to the water is 10-15: 1000, and the pH value is 7.0-7.5;
(3) solid state fermentation: inoculating the seed solution into a first solid fermentation medium for fermentation, and culturing for 4-5 days at 55-60 ℃ to obtain a fermentation product of the thermophilic actinomyces, wherein the formula of the solid fermentation medium is to use wheat bran as a carbon source and a fermentation substrate, add 2-3% of peptone, 1.5-2.0% of NaCl and 0.4-0.5% of nutrient solution, and have an initial pH of 7.0-7.5.
Further, the nutrient solution consists of KNO3, K2HPO4, MgSO4 and FeSO4, and the mass ratio of KNO3, K2HPO4, MgSO4 and FeSO4 is 0.12:0.075:0.15: 0.0001.
Further, the solid-state fermentation of the bacillus licheniformis adopts the following steps:
(1) first-order seed culture: inoculating the bacillus licheniformis strain into a primary seed culture medium from a preserved inclined plane, culturing for 2-3 d, and washing off spores to prepare a primary spore suspension; the formula of the primary seed culture medium comprises 0.5% of peptone, 0.3% of beef extract, 97.2% -97.7% of trace element liquid and 1.5% -2% of agar, the pH is adjusted to be 7.0-8.0, and the trace element liquid comprises CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O, MnSO4.H2O, CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O and MnSO4.H2O, and the mass ratio of the components is 0.5: 0.2: 3: 0.1: 0.1;
(2) secondary seed culture: coating the primary spore suspension on a secondary seed culture medium for culturing for 2-3 d, and washing off spores to prepare secondary spore suspension; the secondary seed culture medium is the same as the primary seed culture medium;
(3) solid state fermentation: inoculating the secondary spore suspension into a second solid state fermentation culture medium according to the inoculation amount of 103-104 cfu/g matrix, and culturing for 5-6 days, wherein the solid state fermentation temperature is 60-65 ℃, and the initial pH value is 7.0-8.0, so as to obtain a bacillus licheniformis fermentation product; the second solid fermentation medium is prepared by compounding corn flour, soybean flour and rice hull into a solid fermentation substrate according to a ratio of 32:3:65, dissolving CaCl2, KH2PO4, (NH4)2HPO4, MgSO4 and MnSO4 according to a ratio of 0.04:0.075:0.05:0.15:0.01, and adding the dissolved substances into the solid fermentation substrate to form an inorganic salt nutrient solution with a final concentration of 1.0%.
The invention has the beneficial effects that:
1. according to the invention, the dry toilet excrement which is difficult to utilize and has heavy peculiar smell is fermented into a treated product with the pathogenic microorganism excrement coliform flora content meeting the harmless requirement of the organic fertilizer, the peculiar smell is reduced, and the total amount is reduced.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below.
Example 1: a compound microbial agent for treating excrement comprises thermophilic actinomycetes and bacillus licheniformis, wherein the thermophilic actinomycetes is preserved in China general microbiological culture Collection center (CGMCC) at 11-09.2020, and the preservation number is CGMCC No. 21143; the bacillus licheniformis is preserved in China general microbiological culture Collection center (CGMCC) at 09.11.2020, with the preservation number of CGMCC No. 21144; the compounding ratio of the thermophilic actinomyces and the bacillus licheniformis is 1:5, and the effective viable count of the thermophilic actinomyces is more than or equal to 2 multiplied by 108cfu/g, the effective viable count of the bacillus licheniformis is more than or equal to 2 multiplied by 108 cfu/g。
Example 2: a compound microbial agent for treating excrement comprises thermophilic actinomycetes and bacillus licheniformis, wherein the thermophilic actinomycetes is preserved in China general microbiological culture Collection center (CGMCC) at 11-09.2020, and the preservation number is CGMCC No. 21143; the bacillus licheniformis is preserved in China general microbiological culture Collection center (CGMCC) at 09.11.2020, with the preservation number of CGMCC No. 21144; the compounding ratio of the thermophilic actinomyces and the bacillus licheniformis is 1:6, and the effective viable count of the thermophilic actinomyces is more than or equal to 2 multiplied by 108cfu/g, the effective viable count of the bacillus licheniformis is more than or equal to 2 multiplied by 108 cfu/g。
Example 3: a compound microbial agent for treating excrement comprises thermophilic actinomycetes and bacillus licheniformis, wherein the thermophilic actinomycetes is preserved in China general microbiological culture Collection center (CGMCC) at 11-09.2020, and the preservation number is CGMCC No. 21143; the bacillus licheniformis is preserved in China general microbiological culture Collection center (CGMCC) at 09.11.2020, with the preservation number of CGMCC No. 21144; the compounding ratio of the thermophilic actinomyces and the bacillus licheniformis is 1:7, and the effective viable count of the thermophilic actinomyces is more than or equal to 2 multiplied by 108cfu/g, the effective viable count of the bacillus licheniformis is more than or equal to 2 multiplied by 108 cfu/g。
Example 4: a compound microbial preparation for treating excrement is used for biological treatment of excrement.
Example 5: one kind is usedThe compounding method of the compound microbial agent for treating excrement comprises the following steps: solid state fermentation: respectively carrying out solid state fermentation on the thermophilic actinomyces and the bacillus licheniformis; and (3) drying: drying the solid fermented thermophilic actinomycetes and bacillus licheniformis at the temperature of 70 ℃, and obtaining thermophilic actinomycetes products and bacillus licheniformis products with the water content of less than 10 percent after drying; mixing and crushing: mixing the thermophilic actinomyces product and the bacillus licheniformis product according to the proportion of 1:5 to form a mixed product, crushing the mixed product, and adding an auxiliary material according to the proportion that the bacteria content meets the requirement to obtain a finished product, wherein the auxiliary material is 150-mesh bentonite. The content of the fresh substrate of the thermophilic actinomycete product is 1.0 multiplied by 1010cfu/g, the bacteria content in the fresh substrate of the fermentation product of the bacillus licheniformis is 2.5 multiplied by 1010 cfu/g。
Solid-state fermentation and shake-flask culture of thermophilic actinomycetes: inoculating the thermophilic actinomycete strain into a shake flask culture medium from a preserved inclined plane, standing, and performing activated culture for 14 hours at the temperature of 55 ℃; the formula of the shake flask culture medium comprises peptone, glucose and water, wherein the mass ratio of the peptone to the glucose to the water is 25:15:1000, and the pH value is 7.0; seed culture: inoculating the thermophilic actinomycetes in the shake flask culture medium into a seed culture medium according to the inoculation amount of 2.0%, and culturing for 12 hours at the rotation speed of 30 r/min and the temperature of 55 ℃ to obtain a seed solution; counting the thermophilic actinomycetes in the seed liquid, wherein the formula of a seed culture medium comprises peptone, beef extract and water, the mass ratio of the peptone to the beef extract to the water is 10:10:1000, and the pH value is 7.0; solid state fermentation: inoculating the seed solution into a first solid state fermentation culture medium for fermentation, and culturing for 5 days at 55 ℃ to obtain a fermentation product of the thermophilic actinomycete, wherein the formula of the solid state fermentation culture medium comprises wheat bran serving as a carbon source and a fermentation substrate, 2% of peptone, 1.5% of NaCl and 0.4% of nutrient solution with the initial pH of 7.0, the nutrient solution consists of KNO3, K2HPO4, MgSO4 and FeSO4, and the mass ratio of KNO3, K2HPO4, MgSO4 and FeSO4 is 0.12:0.075:0.15: 0.0001.
B, solid-state fermentation of the bacillus licheniformis, primary seed culture: inoculating the bacillus licheniformis strain from the preserved slant to a primary seed culture medium for culturing for 2 d, and washing off spores to prepare a primary spore suspension; the first-level seed culture medium is prepared from 0.5% peptone, 0.3% beef extract, 97.7% trace element liquid and 1.5% agar, the pH is adjusted to 7.0, and the trace element liquid comprises CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O, MnSO4.H2O, CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O and MnSO4.H2O, and the mass ratio of the components is 0.5: 0.2: 3: 0.1: 0.1; secondary seed culture: coating the primary spore suspension on a secondary seed culture medium for culturing for 2 d, and washing off spores to prepare a secondary spore suspension; the secondary seed culture medium is the same as the primary seed culture medium. Solid state fermentation: inoculating the secondary spore suspension into a second solid state fermentation culture medium according to the inoculation amount of 103 cfu/g matrix, and culturing for 5d, wherein the solid state fermentation temperature is 65 ℃, and the initial pH value is 7.0, so as to obtain a bacillus licheniformis fermentation product; the second type solid fermentation medium is prepared by compounding corn flour, soybean flour and rice hull into a solid fermentation substrate according to a ratio of 32:3:65, dissolving CaCl2, KH2PO4, (NH4)2HPO4, MgSO4 and MnSO4 according to a ratio of 0.04:0.075:0.05:0.15:0.01, and adding the dissolved substances into the solid fermentation substrate to form an inorganic salt nutrient solution with a final concentration of 1.0%.
Example 6: a compound method of a compound microbial agent for excrement treatment comprises the following steps: solid state fermentation: respectively carrying out solid state fermentation on the thermophilic actinomyces and the bacillus licheniformis; and (3) drying: drying the solid fermented thermophilic actinomycetes and bacillus licheniformis at the temperature of 70 ℃, and obtaining thermophilic actinomycetes products and bacillus licheniformis products with the water content of less than 10 percent after drying; mixing and crushing: mixing the thermophilic actinomyces product and the bacillus licheniformis product according to the proportion of 1:6 to form a mixed product, crushing the mixed product, and adding an auxiliary material according to the proportion that the bacteria content meets the requirement to obtain a finished product, wherein the auxiliary material is 300-mesh bentonite. The content of the fresh substrate of the thermophilic actinomycete product is 1.0 multiplied by 1010cfu/g, the bacteria content in the fresh substrate of the fermentation product of the bacillus licheniformis is 2.5 multiplied by 1010 cfu/g。
Solid-state fermentation and shake-flask culture of thermophilic actinomycetes: inoculating the thermophilic actinomycete strain into a shake flask culture medium from a preserved inclined plane, standing, and performing activated culture for 13 hours at the temperature of 57 ℃; the formula of the shake flask culture medium comprises peptone, glucose and water, wherein the mass ratio of the peptone to the glucose to the water is 25:15:1000, and the pH value is 7.2; seed culture: inoculating the thermophilic actinomycetes in the shake flask culture medium into a seed culture medium according to the inoculation amount of 2.5%, and culturing for 11 h at the rotation speed of 45 r/min and the temperature of 57 ℃ to obtain a seed solution; counting the thermophilic actinomycetes in the seed liquid, wherein the formula of a seed culture medium comprises peptone, beef extract and water, the mass ratio of the peptone to the beef extract to the water is 12:12:1000, and the pH value is 7.0; solid state fermentation: inoculating the seed solution into a first solid state fermentation culture medium for fermentation, and culturing for 4.5 days at 57 ℃ to obtain a fermentation product of the Thermoactinomyces, wherein the formula of the solid state fermentation culture medium comprises wheat bran serving as a carbon source and a fermentation substrate, and is added with 2.5% of peptone, 1.8% of NaCl and 0.45% of nutrient solution, the initial pH is 7.2, the nutrient solution consists of KNO3, K2HPO4, MgSO4 and FeSO4, and the mass ratio of KNO3, K2HPO4, MgSO4 and FeSO4 is 0.12:0.075:0.15: 0.0001.
B, solid-state fermentation of the bacillus licheniformis, primary seed culture: inoculating the Bacillus licheniformis strain from the preserved slant to a primary seed culture medium for culturing for 2.5 d, and washing off spores to obtain a primary spore suspension; the first-level seed culture medium is prepared from 0.5% peptone, 0.3% beef extract, 97.5% trace element liquid and 1.7% agar, the pH is adjusted to 7.5, and the trace element liquid comprises CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O, MnSO4.H2O, CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O and MnSO4.H2O, and the mass ratio of the components is 0.5: 0.2: 3: 0.1: 0.1; secondary seed culture: coating the primary spore suspension on a secondary seed culture medium for culturing for 2.5 d, and washing off spores to prepare secondary spore suspension; the secondary seed culture medium is the same as the primary seed culture medium. Solid state fermentation: inoculating the secondary spore suspension into a second solid state fermentation culture medium according to the inoculation amount of 103.5 cfu/g matrix, and culturing for 5.5d, wherein the solid state fermentation temperature is 63 ℃, and the initial pH value is 7.5, so as to obtain a bacillus licheniformis fermentation product; the second solid fermentation medium is prepared by compounding corn flour, soybean flour and rice hull into a solid fermentation substrate according to a ratio of 32:3:65, dissolving CaCl2, KH2PO4, (NH4)2HPO4, MgSO4 and MnSO4 according to a ratio of 0.04:0.075:0.05:0.15:0.01, and adding the dissolved substances into the solid fermentation substrate to form an inorganic salt nutrient solution with a final concentration of 1.0%.
Example 7: compound microbial agent for excrement treatmentThe preparation method comprises the following steps: solid state fermentation: respectively carrying out solid state fermentation on the thermophilic actinomyces and the bacillus licheniformis; and (3) drying: drying the solid fermented thermophilic actinomycetes and bacillus licheniformis at the temperature of 70 ℃, and obtaining thermophilic actinomycetes products and bacillus licheniformis products with the water content of less than 10 percent after drying; mixing and crushing: mixing the thermophilic actinomyces product and the bacillus licheniformis product according to the proportion of 1:7 to form a mixed product, crushing the mixed product, and adding an auxiliary material according to the proportion that the bacteria content meets the requirement to obtain a finished product, wherein the auxiliary material is 400-mesh bentonite; the content of the fresh substrate of the thermophilic actinomycete product is 1.0 multiplied by 1010cfu/g, the bacteria content in the fresh substrate of the fermentation product of the bacillus licheniformis is 2.5 multiplied by 1010 cfu/g。
Solid-state fermentation and shake-flask culture of thermophilic actinomycetes: inoculating the thermophilic actinomycete strain into a shake flask culture medium from a preserved inclined plane, standing, and performing activated culture for 12 hours at the temperature of 60 ℃; the formula of the shake flask culture medium comprises peptone, glucose and water, wherein the mass ratio of the peptone to the glucose to the water is 25:15:1000, and the pH value is 7.5; seed culture: inoculating the thermophilic actinomycetes in the shake flask culture medium into a seed culture medium according to the inoculation amount of 3.0%, and culturing for 10 hours at the rotating speed of 40 r/min and the temperature of 60 ℃ to obtain a seed solution; counting the thermophilic actinomycetes in the seed liquid, wherein the formula of a seed culture medium comprises peptone, beef extract and water, the mass ratio of the peptone to the beef extract to the water is 15:15:1000, and the pH value is 7.5; solid state fermentation: inoculating the seed solution into a first solid state fermentation culture medium for fermentation, and culturing for 4 days at 60 ℃ to obtain a fermentation product of the Thermoactinomyces, wherein the formula of the solid state fermentation culture medium is to use wheat bran as a carbon source and a fermentation substrate, 3% of peptone, 2.0% of NaCl and 0.5% of nutrient solution are added, the initial pH is 7.5, the nutrient solution consists of KNO3, K2HPO4, MgSO4 and FeSO4, and the mass ratio of KNO3, K2HPO4, MgSO4 and FeSO4 is 0.12:0.075:0.15: 0.0001.
B, solid-state fermentation of the bacillus licheniformis, primary seed culture: inoculating the bacillus licheniformis strain from the preserved slant to a primary seed culture medium for culturing for 3 d, and washing off spores to prepare a primary spore suspension; the formula of the primary seed culture medium comprises 0.5% of peptone, 0.3% of beef extract, 97.2% of trace element liquid and 2% of agar, the pH is adjusted to 8.0, and the trace element liquid comprises CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O, MnSO4.H2O, CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O and MnSO4.H2O, and the mass ratio of the CaCl2.2O to the K2HPO4 to the NH4 to the 2HPO4 to the MgSO4.7H2O to the MnSO4.H: 0.2: 3: 0.1: 0.1; secondary seed culture: coating the primary spore suspension on a secondary seed culture medium for culturing for 3 d, and washing off spores to prepare a secondary spore suspension; the secondary seed culture medium is the same as the primary seed culture medium. Solid state fermentation: inoculating the secondary spore suspension into a second solid state fermentation culture medium according to the inoculation amount of 104 cfu/g matrix, and culturing for 6d, wherein the solid state fermentation temperature is 60 ℃, and the initial pH value is 8.0, so as to obtain a bacillus licheniformis fermentation product; the second type solid fermentation medium is prepared by compounding corn flour, soybean flour and rice hull into a solid fermentation substrate according to a ratio of 32:3:65, dissolving CaCl2, KH2PO4, (NH4)2HPO4, MgSO4 and MnSO4 according to a ratio of 0.04:0.075:0.05:0.15:0.01, and adding the dissolved substances into the solid fermentation substrate to form an inorganic salt nutrient solution with a final concentration of 1.0%.
The described embodiments are only some embodiments of the invention, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Claims (9)
1. A compound microbial agent for treating excrement is characterized by comprising thermophilic actinomycetes and bacillus licheniformis, wherein the thermophilic actinomycetes is preserved in China general microbiological culture Collection center (CGMCC) at 11/09/2020 with the preservation number of CGMCC No. 21143; the bacillus licheniformis is preserved in China general microbiological culture Collection center (CGMCC) at 09.11.2020, with the preservation number of CGMCC No. 21144; the compounding ratio of the thermophilic actinomyces to the bacillus licheniformis is 1: 5-7.
2. The complex microbial inoculant for excrement treatment according to claim 1, wherein the effective viable count of said Thermoactinomyces is 2 x 10 or more8cfu/g, the effective viable count of the bacillus licheniformis is more than or equal to 2 multiplied by 108 cfu/g。
3. The complex microbial inoculant for fecal management according to claim 1 applied to the biological treatment of fecal excrement.
4. A compounding method of a compound microbial agent for excrement treatment is characterized by comprising the following steps:
(1) solid state fermentation: respectively carrying out solid state fermentation on the thermophilic actinomyces and the bacillus licheniformis;
(2) and (3) drying: drying the solid fermented thermophilic actinomycetes and bacillus licheniformis at the temperature of 70 ℃, and obtaining thermophilic actinomycetes products and bacillus licheniformis products with the water content of less than 10 percent after drying;
(3) mixing and crushing: mixing the thermophilic actinomyces product and the bacillus licheniformis product according to the proportion of 1: 5-7 to form a mixed product, crushing the mixed product, and adding an auxiliary material according to the proportion of the bacteria content meeting the requirement to obtain a finished product, wherein the auxiliary material is 150-400 meshes of bentonite.
5. The method for compounding a complex microbial inoculant for excrement treatment according to claim 4, wherein the bacteria content in the fresh substrate of the Thermoactinomyces thermophilus product is 1.0 x 1010cfu/g, the bacteria content in the fresh substrate of the fermentation product of the bacillus licheniformis is 2.5 multiplied by 1010 cfu/g。
6. The method for compounding a complex microbial inoculant for excrement treatment according to claim 4, wherein the bentonite is 300 meshes.
7. The method for compounding the composite microbial inoculant for excrement treatment according to claim 4, wherein the solid-state fermentation of the thermophilic actinomyces comprises the following steps:
(1) and (3) shake flask culture: inoculating the thermophilic actinomycete strain into a shake flask culture medium from a preserved inclined plane, standing, and performing activated culture for 12-14 h at the temperature of 55-60 ℃; the formula of the shake flask culture medium comprises peptone, glucose and water, wherein the mass ratio of the peptone to the glucose to the water is 25:15:1000, and the pH value is 7.0-7.5;
(2) seed culture: inoculating the thermophilic actinomyces in the shake flask culture medium into a seed culture medium according to the inoculation amount of 2.0-3.0%, and culturing at the rotation speed of 30-40 r/min and the temperature of 55-60 ℃ for 10-12 h to obtain a seed solution; counting the thermophilic actinomycetes in the seed liquid, wherein the formula of the seed culture medium comprises peptone, beef extract and water, the mass ratio of the peptone to the beef extract to the water is 10-15: 1000, and the pH value is 7.0-7.5;
(3) solid state fermentation: inoculating the seed solution into a first solid fermentation medium for fermentation, and culturing for 4-5 days at 55-60 ℃ to obtain a fermentation product of the thermophilic actinomyces, wherein the formula of the solid fermentation medium is to use wheat bran as a carbon source and a fermentation substrate, add 2-3% of peptone, 1.5-2.0% of NaCl and 0.4-0.5% of nutrient solution, and have an initial pH of 7.0-7.5.
8. The method for compounding the compound microbial inoculant for the treatment of excrement according to claim 7, wherein the nutrient solution consists of KNO3, K2HPO4, MgSO4 and FeSO4, and the mass ratio of KNO3, K2HPO4, MgSO4 and FeSO4 is 0.12:0.075:0.15: 0.0001.
9. The compounding method of the composite microbial agent for excrement treatment according to claim 4, wherein the Bacillus licheniformis solid state fermentation adopts the following steps:
(1) first-order seed culture: inoculating the bacillus licheniformis strain into a primary seed culture medium from a preserved inclined plane, culturing for 2-3 d, and washing off spores to prepare a primary spore suspension; the formula of the primary seed culture medium comprises 0.5% of peptone, 0.3% of beef extract, 97.2% -97.7% of trace element liquid and 1.5% -2% of agar, the pH is adjusted to be 7.0-8.0, and the trace element liquid comprises CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O, MnSO4.H2O, CaCl2.2H2O, K2HPO4, (NH4)2HPO4, MgSO4.7H2O and MnSO4.H2O, and the mass ratio of the components is 0.5: 0.2: 3: 0.1: 0.1;
(2) secondary seed culture: coating the primary spore suspension on a secondary seed culture medium for culturing for 2-3 d, and washing off spores to prepare secondary spore suspension; the secondary seed culture medium is the same as the primary seed culture medium;
(3) solid state fermentation: inoculating the secondary spore suspension into a second solid state fermentation culture medium according to the inoculation amount of 103-104 cfu/g matrix, and culturing for 5-6 days, wherein the solid state fermentation temperature is 60-65 ℃, and the initial pH value is 7.0-8.0, so as to obtain a bacillus licheniformis fermentation product; the second solid fermentation medium is prepared by compounding corn flour, soybean flour and rice hull into a solid fermentation substrate according to a ratio of 32:3:65, dissolving CaCl2, KH2PO4, (NH4)2HPO4, MgSO4 and MnSO4 according to a ratio of 0.04:0.075:0.05:0.15:0.01, and adding the dissolved substances into the solid fermentation substrate to form an inorganic salt nutrient solution with a final concentration of 1.0%.
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