CN112063557A - Perishable garbage low-temperature phase change water-producing degradation microbial inoculum and microbial liquid preparation method - Google Patents
Perishable garbage low-temperature phase change water-producing degradation microbial inoculum and microbial liquid preparation method Download PDFInfo
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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
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/341—Consortia of bacteria
Abstract
The invention relates to a degrading microbial inoculum, in particular to a perishable garbage low-temperature phase-change water-producing degrading microbial inoculum and a preparation method of microbial inoculum, and belongs to the technical field of microorganisms. The microbial fertilizer comprises a compound microorganism, wherein the compound microorganism comprises the following raw materials in parts by mass: 5-6 parts of clostridium difficile, 5-6 parts of ephedra, 9-11 parts of geobacillus, 9-11 parts of acetobacter, 34-40 parts of clostridium, 24-30 parts of clostridium thiolyticum and 5-6 parts of bacteroides. The method comprises the following steps: preparing microbial fermentation liquid → freeze-dried powder, microbial liquid or mixture of microbial liquid and solid auxiliary material. The treated wastewater reaches the discharge standard, and the functions of reducing garbage and protecting the environment are achieved.
Description
Technical Field
The invention relates to a degrading microbial inoculum, in particular to a perishable garbage low-temperature phase-change water-producing degrading microbial inoculum and a preparation method of microbial inoculum, and belongs to the technical field of microorganisms.
Background
With the rapid development of social economy and the acceleration of urbanization in China, the yield of municipal domestic garbage is continuously increased, and the perishable garbage in the domestic garbage accounts for about half. Perishable rubbish, also can wet rubbish or kitchen garbage, the kitchen waste that produces in the production processes such as food and beverage operator, unit dining room is generally referred to and the perishable rubbish that produces in the family life mainly includes: leftovers, stems and leaves, meat entrails, husks and peels, and the like. The perishable garbage contains a large amount of starch, protein, grease, plant fiber and the like, is rich in nutrition, and if the perishable garbage is not treated well, the perishable garbage is easy to cause mass propagation of harmful organisms such as germs, mosquitoes and the like, and threatens human health and urban ecological environment. At present, the perishable garbage is treated mainly by a landfill method, an incineration method, a composting method and a high-temperature anaerobic digestion method. Although the landfill method, the incineration method and the composting method are simpler to treat, the landfill method, the incineration method and the composting method are easy to cause secondary pollution to soil, water and air. The high-temperature anaerobic digestion method is a more advanced treatment mode, but the method has the defects of high technical threshold, large equipment capital investment and overhigh operation energy consumption cost, and is not suitable for all situations.
Disclosure of Invention
The invention mainly solves the defects in the prior art, and provides a perishable waste low-temperature phase-change water degradation microbial inoculum and a microbial liquid preparation method, wherein perishable waste is crushed and then put into kitchen waste treatment equipment, then a low-temperature oxygen-consuming microbial inoculum is added, in the process, swill carrying the microbial inoculum enters a sewage treatment system to be converted into wastewater which reaches the standard and is discharged, and the perishable waste is converted into harmless water and carbon dioxide under the decomposition action of microorganisms.
The technical problem of the invention is mainly solved by the following technical scheme:
the perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises compound microorganisms, wherein the compound microorganisms comprise the following raw materials in parts by mass:
5-6 parts of clostridium difficile, 5-6 parts of ephedra, 9-11 parts of geobacillus, 9-11 parts of acetobacter, 34-40 parts of clostridium, 24-30 parts of clostridium thiolyticum and 5-6 parts of bacteroides.
Preferably, the compound microorganism comprises the following raw materials in parts by mass: 5 parts of clostridium difficile, 5 parts of ephedra difficile, 9 parts of geobacillus, 11 parts of acetobacter, 34 parts of clostridium, 30 parts of clostridium thiolyticum and 6 parts of bacteroides.
Preferably, the compound microorganism comprises the following raw materials in parts by mass: 5 parts of clostridium difficile, 6 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 35 parts of clostridium, 28 parts of clostridium thiolyticum and 6 parts of bacteroides.
Preferably, the compound microorganism comprises the following raw materials in parts by mass: 6 parts of clostridium difficile, 5 parts of ephedra difficile, 11 parts of geobacillus, 9 parts of acetobacter, 38 parts of clostridium, 26 parts of clostridium thiogenes and 5 parts of bacteroides.
Preferably, the compound microorganism comprises the following raw materials in parts by mass: 6 parts of clostridium difficile, 5 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 40 parts of clostridium, 24 parts of clostridium thiolyticum and 5 parts of bacteroides.
Preferably, the compound microorganism exists in the form of freeze-dried powder, microorganism liquid or a mixture of the microorganism liquid and solid auxiliary materials.
Preferably, the weight ratio of the solid auxiliary material to the compound microorganism is 4: 1.
A preparation method of a perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
culturing irregular clostridium on an intensified clostridium culture medium at 25 ℃ under the condition of slant culture, then culturing by secondary seed liquid, fermenting and culturing until the concentration of clostridium reaches 1-2 multiplied by 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 1-2 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth culture medium at 28-30 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: culturing Acetobacter on acetic acid bacteria culture medium at 30-32 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until bacteria count reaches 1-2 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 28-30 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
6) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes in Clostridium enrichment medium at 36-38 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
7) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 30-35 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 1-2 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
Preferably, the solid auxiliary material is prepared from peptone and beta dextrin according to the ratio of 1: 1 are mixed.
Preferably, the reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 1000mL of distilled water, and adjusting the pH value to be 6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.0-7.2;
mixing the above formulas, adding distilled water, dissolving and mixing, adjusting pH to 7.0-7.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate, 1000mL of distilled water and pH 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
The english trade name of clostridium difficile is: clostridium irregular;
the English trade name of the irregular Ephedra distachya is: asaccharospora irregular;
the english trade name of the genus terrestris is: terrispobacter petrilerarius;
the english trade name of acetobacter is: acetobacter sp;
the english trade name of clostridium is: clostridium sp.;
the english trade name of clostridium thiogenes is: clostridium sulfoxigenes;
the English trade name of Bacteroides is: bacterioides xylanolyticus sp;
the English commodity of the reinforced clostridia culture medium is simply called as: RCM;
the English commodity of the thioglycolate fluid medium is simply called: FT;
the english trade name of resazurin is: resazurin;
the microbial inoculum prepared by fermentation has the characteristics of special fermentation smell, stable state, cooperative symbiosis, no pathogenic bacteria, small application addition amount, safety, environmental protection and the like. The method can quickly play a role in a low-temperature oxygen consumption environment when put into a bin of perishable garbage treatment equipment, the garbage reduction rate exceeds 90 percent, and perishable garbage with high fat, high salt and high heat is converted into carbon dioxide and water which are environment-friendly. The microbial inoculum can greatly save the energy consumption of equipment and has no secondary pollution in operation.
The invention provides a perishable garbage low-temperature phase change water-producing degradation microbial inoculum and a preparation method of microbial inoculum, and the treated wastewater reaches the discharge standard, thereby playing the functions of reducing garbage and protecting environment.
Detailed Description
The technical scheme of the invention is further specifically described by the following embodiments.
Example 1: the perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises compound microorganisms, wherein the compound microorganisms comprise the following raw materials in parts by mass:
5 parts of clostridium difficile, 5 parts of ephedra difficile, 9 parts of geobacillus, 11 parts of acetobacter, 34 parts of clostridium, 30 parts of clostridium thiolyticum and 6 parts of bacteroides.
The compound microorganism exists in the form of freeze-dried powder, microorganism bacterium liquid or mixture of the microorganism bacterium liquid and solid auxiliary materials.
The weight ratio of the solid auxiliary materials to the composite microorganisms is 4: 1.
A preparation method of a perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
clostridium difficile in strongPerforming slant culture and secondary seed liquid culture on a Clostridium difficile culture medium at the ambient temperature of 25 ℃, and performing fermentation culture until the concentration of the bacteria reaches 1 × 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 1 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth at 28 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 3 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: culturing Acetobacter on acetic acid bacteria culture medium at 30 deg.C, performing first-stage slant culture, inoculating to triangular flask, performing shaking second-stage liquid culture until bacteria count reaches 1 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 28 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3 × 108Per mL;
7) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes in Clostridium enrichment medium at 36 deg.C, inoculating to triangular flask, performing oscillation second-stage liquid culture until the number of bacteria reaches 3 × 108Per mL;
8) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 30 deg.C, inoculating into triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 1 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
The solid auxiliary materials comprise peptone and beta dextrin according to the proportion of 1: 1 are mixed.
The reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 1000mL of distilled water, and adjusting the pH value to be 6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.0;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.0, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate, 1000mL of distilled water and pH 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
Example 2:
the perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises compound microorganisms, wherein the compound microorganisms comprise the following raw materials in parts by mass:
5 parts of clostridium difficile, 6 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 35 parts of clostridium, 28 parts of clostridium thiolyticum and 6 parts of bacteroides.
The compound microorganism exists in the form of freeze-dried powder, microorganism bacterium liquid or mixture of the microorganism bacterium liquid and solid auxiliary materials.
The weight ratio of the solid auxiliary materials to the composite microorganisms is 4: 1.
A preparation method of a perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
culturing irregular clostridium on an intensified clostridium culture medium at 25 ℃ under the condition of slant culture, then culturing by secondary seed liquid, fermenting and culturing until the concentration of clostridium reaches 2 multiplied by 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 2 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth at 29 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: culturing Acetobacter on acetic acid bacteria culture medium at 31 deg.C, performing first-stage slant culture, inoculating into triangular flask, performing shaking second-stage liquid culture until bacteria count reaches 2 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 29 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
8) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes on Clostridium enrichment medium at 37 deg.C, inoculating to triangular flask, performing oscillation second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
9) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 33 deg.C, inoculating into triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 2 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
The solid auxiliary materials comprise peptone and beta dextrin according to the proportion of 1: 1 are mixed.
The reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 1000mL of distilled water, and adjusting the pH value to be 6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.1;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate and 1000mL of distilled water, and adjusting the pH value to 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
Example 3:
the perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises compound microorganisms, wherein the compound microorganisms comprise the following raw materials in parts by mass:
6 parts of clostridium difficile, 5 parts of ephedra difficile, 11 parts of geobacillus, 9 parts of acetobacter, 38 parts of clostridium, 26 parts of clostridium thiogenes and 5 parts of bacteroides.
The compound microorganism exists in the form of freeze-dried powder, microorganism bacterium liquid or mixture of the microorganism bacterium liquid and solid auxiliary materials.
The weight ratio of the solid auxiliary materials to the composite microorganisms is 4: 1.
A preparation method of a perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
culturing irregular clostridium on an intensified clostridium culture medium at 25 ℃ under the condition of slant culture, then culturing by secondary seed liquid, fermenting and culturing until the concentration of clostridium reaches 2 multiplied by 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 2 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth at 30 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: culturing Acetobacter on acetic acid bacteria culture medium at 32 deg.C, performing first-stage slant culture, inoculating to triangular flask, performing shake second-stage liquid culture until bacteria count reaches 2 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 30 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
9) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes on Clostridium enrichment medium at 38 deg.C, inoculating to triangular flask, performing oscillation second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
10) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 35 deg.C, inoculating into triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 2 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
The solid auxiliary materials comprise peptone and beta dextrin according to the proportion of 1: 1 are mixed.
The reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 1000mL of distilled water, and adjusting the pH value to be 6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate and 1000mL of distilled water, and adjusting the pH value to 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
Example 4:
the perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises compound microorganisms, wherein the compound microorganisms comprise the following raw materials in parts by mass:
6 parts of clostridium difficile, 5 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 40 parts of clostridium, 24 parts of clostridium thiolyticum and 5 parts of bacteroides.
The compound microorganism exists in the form of freeze-dried powder, microorganism bacterium liquid or mixture of the microorganism bacterium liquid and solid auxiliary materials.
The weight ratio of the solid auxiliary materials to the composite microorganisms is 4: 1.
A preparation method of a perishable garbage low-temperature phase change water-producing degradation microbial inoculum comprises the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
culturing irregular clostridium on an intensified clostridium culture medium at 25 ℃ under the condition of slant culture, then culturing by secondary seed liquid, fermenting and culturing until the concentration of clostridium reaches 2 multiplied by 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 2 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth at 30 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: culturing Acetobacter on acetic acid bacteria culture medium at 31 deg.C, performing first-stage slant culture, inoculating into triangular flask, performing shaking second-stage liquid culture until bacteria count reaches 1-2 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 29 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
10) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes in Clostridium enrichment medium at 36 deg.C, inoculating to triangular flask, performing oscillation second-stage liquid culture until the number of bacteria reaches 4 × 108Per mL;
11) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 35 deg.C, inoculating into triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 1 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
The solid auxiliary materials comprise peptone and beta dextrin according to the proportion of 1: 1 are mixed.
The reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 1000mL of distilled water, and adjusting the pH value to be 6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate, 1000mL of distilled water and pH 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
Claims (10)
1. The low-temperature phase change water-producing and degrading microbial inoculum for perishable garbage is characterized in that: the microbial fertilizer comprises a compound microorganism, wherein the compound microorganism comprises the following raw materials in parts by mass:
5-6 parts of clostridium difficile, 5-6 parts of ephedra, 9-11 parts of geobacillus, 9-11 parts of acetobacter, 34-40 parts of clostridium, 24-30 parts of clostridium thiolyticum and 5-6 parts of bacteroides.
2. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 1, characterized in that: the composite microorganism comprises the following raw materials in parts by mass: 5 parts of clostridium difficile, 5 parts of ephedra difficile, 9 parts of geobacillus, 11 parts of acetobacter, 34 parts of clostridium, 30 parts of clostridium thiolyticum and 6 parts of bacteroides.
3. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 1, characterized in that: the composite microorganism comprises the following raw materials in parts by mass: 5 parts of clostridium difficile, 6 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 35 parts of clostridium, 28 parts of clostridium thiolyticum and 6 parts of bacteroides.
4. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 1, characterized in that: the composite microorganism comprises the following raw materials in parts by mass: 6 parts of clostridium difficile, 5 parts of ephedra difficile, 11 parts of geobacillus, 9 parts of acetobacter, 38 parts of clostridium, 26 parts of clostridium thiogenes and 5 parts of bacteroides.
5. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 1, characterized in that: the composite microorganism comprises the following raw materials in parts by mass: 6 parts of clostridium difficile, 5 parts of ephedra difficile, 10 parts of geobacillus, 10 parts of acetobacter, 40 parts of clostridium, 24 parts of clostridium thiolyticum and 5 parts of bacteroides.
6. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 1, characterized in that: the compound microorganism exists in the form of freeze-dried powder, microorganism bacterium liquid or mixture of the microorganism bacterium liquid and solid auxiliary materials.
7. A perishable waste low temperature phase change water degradation microbial inoculum, according to claim 6, characterized in that: the weight ratio of the solid auxiliary materials to the composite microorganisms is 4: 1.
8. The preparation method of the perishable waste low-temperature phase change water degradation microbial inoculum according to claim 1 or 7, which is characterized by comprising the following steps:
preparing a microbial fermentation broth independently:
1) the preparation method of the irregular clostridium microbial fermentation liquor comprises the following steps:
culturing irregular clostridium on an intensified clostridium culture medium at 25 ℃ under the condition of slant culture, then culturing by secondary seed liquid, fermenting and culturing until the concentration of clostridium reaches 1-2 multiplied by 108Per mL;
2) the preparation method of the irregular ephedra fungus microbial fermentation liquor comprises the following steps:
culturing irregular Ephedra sinica Stapf on reinforced Clostridium culture medium at 25 deg.C under slant, culturing with secondary seed liquid, fermenting until the concentration reaches 1-2 × 108Per mL;
3) the preparation method of the fermentation liquor of the Cladosporium microorganisms comprises the following steps:
culturing Cladosporium on nutrient broth culture medium at 28-30 deg.C under first-stage slant, inoculating into triangular flask, and performing shaking second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
4) the preparation method of the acetobacter microorganism fermentation liquor comprises the following steps: acetobacter in AcetobacterPerforming first-stage slant culture at 30-32 deg.C, inoculating to triangular flask, performing shaking second-stage liquid culture until the number of bacteria reaches 1-2 × 108Per mL;
5) the preparation method of the fermentation liquor of the clostridium microorganism comprises the following steps:
performing first-stage slant culture of Clostridium on nutrient broth culture medium at 28-30 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
6) the preparation method of the thioclostridium microbial fermentation liquor comprises the following steps:
performing first-stage slant culture of Clostridium thiogenes in Clostridium enrichment medium at 36-38 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 3-4 × 108Per mL;
7) the preparation method of the bacteroides microorganism fermentation liquor comprises the following steps:
performing first-stage slant culture of Bacteroides on thioglycollate fluid culture medium at 30-35 deg.C, inoculating to triangular flask, performing shake second-stage liquid culture until the number of bacteria reaches 1-2 × 108Per mL;
(II) preparing freeze-dried powder, microbial bacteria liquid or a mixture of the microbial bacteria liquid and solid auxiliary materials:
1) freeze-drying powder preparation:
mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished, and preparing freeze-dried powder according to a conventional method in the field;
2) preparing a microbial liquid:
and (2) mixing various composite microbial fermentation liquids prepared in the step (one) according to the mass ratio of the claim 1 after fermentation is finished to prepare the composite microbial liquid.
3) Preparing a mixture of the microbial liquid and the solid auxiliary materials:
and (2) mixing the various composite microbial fermentation liquids prepared in the step (one) according to the mass percentage ratio after fermentation is finished to prepare a composite microbial liquid, and uniformly mixing 1 part of the composite microbial liquid and 4 parts of solid auxiliary materials to prepare a mixture of the microbial liquid and the solid auxiliary materials.
9. The method for preparing the perishable waste low-temperature phase change water degradation microbial inoculum according to claim 8, wherein the method comprises the following steps: the solid auxiliary materials comprise peptone and beta dextrin according to the proportion of 1: 1 are mixed.
10. The method for preparing the perishable waste low-temperature phase change water degradation microbial inoculum according to claim 8, wherein the method comprises the following steps:
the reinforced clostridium culture medium is prepared by mixing the following raw materials:
10.0g of peptone, 10.0g of beef powder, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride, 1000mL of distilled water and pH6.8 +/-0.1;
mixing the above formula, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.1, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the nutrient broth culture medium is prepared by mixing the following raw materials:
5g of peptone, 30g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and adjusting the pH value to 7.0-7.2;
mixing the above formulas, adding distilled water, dissolving and mixing, adjusting pH to 7.0-7.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the acetic acid bacteria culture medium is prepared by mixing the following raw materials:
100g of glucose, 10g of yeast extract, 20g of calcium carbonate, 1000mL of distilled water and pH 6.8;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8, sterilizing in autoclave under 0.12MPa for 20 min, and cooling;
the clostridium enrichment medium is prepared by mixing the following raw materials:
10.0g of beef powder, 10.0g of monthly peptone, 3.0g of yeast powder, 5.0g of glucose, 1.0g of soluble starch, 5.0g of sodium chloride, 3.0g of sodium acetate, 0.5g of L-cysteine hydrochloride and 0.5g of agar, and adjusting the pH value to be 6.8 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 6.8 + -0.2, sterilizing in autoclave at 0.12MPa for 20 min, and cooling;
the thioglycollate fluid culture medium is prepared by mixing the following raw materials:
trypticase digest casein peptone 15.0g, L-cystine 0.5g, anhydrous glucose 5.0g, yeast extract 5.0g, sodium chloride 2.5g, sodium thioglycolate 0.5g, resazurin 1.0mL with the concentration of 0.1%, distilled water 1000mL, and the pH value is adjusted to 7.1 +/-0.2;
mixing the above formulas, adding distilled water, dissolving and mixing uniformly, adjusting pH to 7.1 + -0.2, sterilizing in autoclave under 0.12MPa for 20 min, and cooling.
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