CN113716985A - Method for exciting organic waste to quickly become thoroughly decomposed and special exciting preparation thereof - Google Patents
Method for exciting organic waste to quickly become thoroughly decomposed and special exciting preparation thereof Download PDFInfo
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- CN113716985A CN113716985A CN202110355181.XA CN202110355181A CN113716985A CN 113716985 A CN113716985 A CN 113716985A CN 202110355181 A CN202110355181 A CN 202110355181A CN 113716985 A CN113716985 A CN 113716985A
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- 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
- C05F17/10—Addition or removal of substances other than water or air to or from the material during the treatment
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- 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
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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
- C12N1/14—Fungi; Culture media therefor
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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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- 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
Abstract
The invention discloses a method for exciting organic waste to be quickly decomposed and a special exciting preparation thereof. The organic waste compost stimulating agent comprises an independently packaged liquid component A and an independently packaged solid component B; the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum; the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacteria agent, 200-600 parts of biochemical fulvic acid raw powder, 300-600 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.00001-0.01 part of auxin. The mass ratio of the liquid component A packaged independently to the solid component B packaged independently is 1: (1.5-4). The organic waste compost exciting preparation provided by the invention can quickly raise compost temperature, increase high-temperature period, accelerate the increase of the microbial quantity of compost, promote compost maturity and shorten fermentation period.
Description
Technical Field
The invention belongs to the technical field of organic waste resource utilization, and particularly relates to a method for exciting organic waste to be quickly decomposed and a special exciting preparation thereof.
Background
The composting is one of important resource ways of organic wastes, most of the domestic organic wastes need to be fermented for 15-20 days when the composting reaches the harmless standard, the fermentation of the formed high-quality organic fertilizer generally needs 45-60 days, and the long period is the main bottleneck of the composting technology. Researches show that the fermentation period can be obviously shortened by adding caramel, polyethylene glycol, medical stone, waste lime, traditional Chinese medicine residues and the like. In comparison, inoculation with microbial decomposing inoculants is an effective and economical process.
The organic waste composting process shortens the composting period and achieves good composting effect, and the key points are how to quickly heat the compost body, and how to maintain the temperature in a high-temperature state for a certain time. At present, most of the existing compost fermentation inoculants can shorten a certain compost starting fermentation time compared with natural compost, but the high-temperature period is too short, the composting is not thorough, and the composting period is still longer.
Disclosure of Invention
The invention aims to provide a method for exciting organic waste to be quickly decomposed and a special exciting preparation thereof.
The invention provides an organic waste compost stimulating preparation, which comprises an independently packaged liquid component A and an independently packaged solid component B;
The independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacteria agent, 200-600 parts of biochemical fulvic acid raw powder, 300-600 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.00001-0.01 part of auxin.
The organic waste compost activating agent may be specifically any one of the following (1) to (5):
(1) comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacteria agent, 200-600 parts of biochemical fulvic acid raw powder, 300 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.00001-0.01 part of auxin;
(2) comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacterium agent, 200-600 parts of biochemical fulvic acid raw powder, 300-600 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.0001 part of auxin;
(3) Comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300-500 parts of phosphate-solubilizing bacterium agent, 300-400 parts of biochemical fulvic acid raw powder, 300 parts of biochar, 0.008-0.01 part of gibberellic acid solid preparation and 0.0001 part of auxin;
(4) comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300 parts of phosphate-solubilizing bacterium agent, 300 parts of biochemical fulvic acid raw powder, 300 parts of biochar, 0.008 part of gibberellic acid solid preparation and 0.0001 part of auxin;
(5) comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 500 parts of phosphate-solubilizing bacterium agent, 400 parts of biochemical fulvic acid raw powder, 300 parts of biochar, 0.01 part of gibberellic acid solid preparation and 0.0001 part of auxin;
In the organic waste compost activating preparation, the mass ratio of the independently packaged liquid component a to the independently packaged solid component B may be 1: (1.5-4), which can be 1: (2-3) and 1: 2 or 1: 3.
in the organic waste compost exciting preparation, the total effective viable count of the mixed zymocyte liquid of the bacillus subtilis and the trichoderma harzianum can be more than or equal to 1000 hundred million CFU/g, such as 1000 hundred million CFU/g.
In the mixed fermentation bacterial liquid of the bacillus subtilis and the trichoderma harzianum, the ratio (CFU ratio) of the bacillus subtilis to the trichoderma harzianum is not less than 10: 1, as 10: 1.
in the organic waste compost exciting preparation, the mixed fermentation bacteria liquid of bacillus subtilis and trichoderma harzianum can be prepared by the following steps: and inoculating the seed solution of the bacillus subtilis and the seed solution of the trichoderma harzianum into a composite fermentation culture medium, and finishing fermentation culture.
Specifically, the composite fermentation medium substrate can be prepared by the following steps: mixing the following components with the balance of water according to the mass percentage of 100 percent: 1.5% of corn flour, 0.5% of corn steep liquor, 5% of corn straw powder, 1.5% of starch, 2% of medium-temperature soybean cake powder, 0.6% of fish meal, and FeSO 4 0.1％，MnSO4 0.1％，KH2PO4 0.1％，MgSO40.05％，CuSO40.05 percent to obtain a mixed solution; regulating and controlling the pH value of the mixed solution to 7.0 to obtain the composite fermentation medium substrate.
Further, the inoculation amount of the seed solution of the bacillus subtilis and the inoculation amount of the seed solution of the trichoderma harzianum can be 5-10 percent, such as 5 percent;
further, the steps of the fermentation culture may be as follows: firstly, fermenting and culturing for 24-48 h (such as 24h or 48h) under the condition that the temperature is 32-35 ℃ (such as 32 ℃) and the stirring speed is 150-180 rpm (such as 180 rpm); then fermenting and culturing for 48-72 h (such as 60-72 h, 60h or 72h) under the condition that the temperature is 25-28 ℃ (such as 26-28 ℃, 26 ℃ or 28 ℃) and the stirring speed is 120-150 rpm (such as 150 rpm).
In the organic waste compost exciting preparation, the phosphate solubilizing bacterium agent can be a bacillus megaterium solid bacterium agent.
In the organic waste compost exciting preparation, the number of effective viable bacteria in the phosphate solubilizing bacteria agent can be more than or equal to 100 hundred million CFU/g, such as 100 hundred million CFU/g.
In the organic waste compost exciting preparation, the biochemical fulvic acid raw powder is a product generated by taking plants such as straws and/or turf as raw materials and performing multi-stage fermentation by using different floras, and specifically can be commercially available biochemical fulvic acid raw powder. In the raw biochemical fulvic acid powder, the mass percentage of the biochemical fulvic acid can be more than or equal to 90%.
In the organic waste compost exciting preparation, the biochar can be straw biochar.
In the organic waste compost exciting preparation, the gibberellic acid solid preparation can be gibberellic acid crystal powder. In the gibberellic acid solid preparation, the content of gibberellic acid in the gibberellic acid solid preparation can be 75% by mass.
In the organic waste compost stimulating agent, the auxin may be indoleacetic acid.
The invention further provides application of the organic waste compost exciting preparation in preparation of products for exciting quick decomposition of organic wastes.
In the above application, the organic waste may be at least one of agricultural organic waste, forestry organic waste and domestic organic waste, such as at least one of crop straw, landscaping grass, livestock and poultry manure, kitchen waste, and the like.
The invention also provides a method for exciting the organic waste to be quickly decomposed by the organic waste exciting preparation, which comprises the following steps: and composting the organic waste added with the exciting agent, and quickly decomposing the organic waste after composting.
In the above method, the amount of the exciting agent added may be 1% to 5%, specifically 3% to 5%, 3% or 5% of the amount of the organic waste.
In the method, the organic waste can be at least one of agricultural organic waste, forestry organic waste and domestic organic waste, such as at least one of crop straws, landscaping grass, livestock and poultry manure, kitchen waste and the like.
The invention has the following beneficial effects:
the organic waste compost exciting preparation provided by the invention can quickly raise compost temperature, increase high-temperature period, accelerate the increase of the microbial quantity of compost, promote compost maturity and shorten fermentation period.
Drawings
FIG. 1 is a temperature profile of different treatments in example 1, wherein T represents the challenge preparation of the present invention and CK represents a commercially available decomposing inoculant.
FIG. 2 is a graph showing the temperature change of different treatments in example 2.
Detailed Description
The organic waste compost stimulating agent provided by the invention comprises an independently packaged liquid component A and an independently packaged solid component B; the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum; the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacteria agent, 200-600 parts of biochemical fulvic acid raw powder, 300-600 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.00001-0.01 part of auxin. The inventive activator can prolong the high temperature period of compost, to accelerate the organic waste to be decomposed, to shorten the composting period.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Bacillus subtilis, ATCC63501 in the following examples;
trichoderma harzianum, ACCC 30371;
bacillus megaterium, having phosphate solubilizing functionality; the product name of the bacillus megaterium solid microbial inoculum purchased from the vigorous biotechnology limited company in the North sea is bacillus megaterium, and the effective viable count is 100 hundred million CFU/g.
The fulvic acid is purchased from Touchfang Songzhi technical limited company, and the product name is biochemical fulvic acid raw powder, English is abbreviated as: BFA; BFA is more than or equal to 90 percent; the pH value is 6-8; the source is as follows: straw and turf.
Biochar was purchased from the environment protection science and technology of south Henan Lize and is named as biochar.
The gibberellic acid solid preparation is purchased from Shanghai Tongri Biotech limited, and has a product name of gibberellic acid, and the content of effective components is as follows: 75%, dosage form: and (4) crystallizing the powder.
Auxin was purchased from Henan Shenyu Biotech Co., Ltd and is available under the product name indoleacetic acid.
Example 1 preparation of priming formulations
Activating bacillus subtilis, and culturing in a seed culture medium: the broth culture medium comprises the following components: 10.0g of beef extract, 10.0g of peptone, 10.0g of glucose, 5.0g of NaCl, 1.0L of distilled water and pH 7.0.
Trichoderma harzianum activation, seed culture medium: the PDA culture medium has the following formula: 1.0L of potato extract, 20.0g of glucose and natural pH. Wherein, the potato extract: removing peel of potato 200g, cutting into small pieces, adding water 1.0L, boiling for 30min, filtering to remove potato pieces, and adding filtrate to 1.0L.
The formula of the composite fermentation medium comprises: 1.5 percent of corn flour, 0.5 percent of corn steep liquor,5% of corn stalk powder, 1.5% of starch, 2% of medium-temperature soybean cake powder, 0.6% of fish meal and FeSO4 0.1％，MnSO4 0.1％，KH2PO4 0.1％，MgSO4 0.05％， CuSO40.05%, make up the filtrate to 1.0L, and adjust the pH to 7.0.
First, preparation method
The challenge formulation was prepared as follows:
1. preparation of the liquid component a packaged separately:
b, bacillus subtilis seed liquid:
activating strains: culturing the bacillus subtilis strain in an activation culture medium for later use;
seed liquid: oscillating the activated bacillus subtilis strain at 37 ℃ for 12h to obtain a bacillus subtilis seed solution after culture;
trichoderma harzianum seed liquid:
activating strains: activating trichoderma harzianum for later use;
seed liquid: shaking the activated trichoderma harzianum strain at 30 ℃ for 24h, and culturing to obtain trichoderma harzianum seed liquid;
mixing zymophyte liquid: performing high-temperature sterilization on a composite fermentation culture medium, then respectively inoculating a bacillus subtilis seed solution and a trichoderma harzianum seed solution according to the inoculation amount of 5% when the temperature is reduced to be below 30 ℃, performing fermentation culture for 48 hours under the conditions that the temperature is 32 ℃ and the stirring speed is 180rpm, and performing fermentation culture for 72 hours under the conditions that the temperature is 26 ℃ and the stirring speed is 150 rpm; after the fermentation culture is finished, concentrating the fermentation liquor until the total effective viable count is more than or equal to 1000 hundred million CFU/g and the ratio of the bacillus subtilis to the trichoderma harzianum is more than or equal to 10:1, thus obtaining the mixed fermentation bacterial liquid.
2. Preparation of separately packaged solid component B:
mixing the following components in parts by weight: 300 parts of bacillus megatherium solid microbial inoculum, 300 parts of biochemical fulvic acid raw powder, 300 parts of charcoal, 0.008 part of gibberellic acid crystal powder and 0.0001 part of indoleacetic acid to obtain an independently packaged solid component B.
Second, formula
This example elicited formulation comprised an individually packaged liquid component a and an individually packaged solid component B;
the mass ratio of the liquid component A packaged independently to the solid component B packaged independently is 1: 2;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum; the total effective viable count of the mixed fermentation bacteria liquid of the bacillus subtilis and the trichoderma harzianum is 1000 hundred million CFU/g, wherein the ratio of the bacillus subtilis to the trichoderma harzianum is 10: 1;
the independently packaged component B consists of the following components in parts by weight:
300 parts of bacillus megatherium solid microbial inoculum, 300 parts of biochemical fulvic acid raw powder, 300 parts of charcoal, 0.008 part of gibberellic acid crystal powder and 0.0001 part of indoleacetic acid.
Third, the effect of use
Vegetable straws are used as main raw materials for carrying out composting tests. The tomato straw is taken from a vegetable greenhouse of agriculture and forestry academy of sciences of Beijing; the garden greening grass is collected from gardens around the Beijing academy of agriculture and forestry; the cow dung is collected from a cattle farm in the ecological agriculture technology garden of China farm machinery institute in the Pingtang district of Beijing. The tomato stems and the lawn grass are crushed and cut into pieces with the length of 3-5 cm. The mass ratio of the raw materials is vegetable straw: grass: and (3) the cow dung is 4:1:2, and the C/N of the mixed material needs to be controlled to be 25-35.
When the composting is started, the humidity is adjusted to about 60 percent. Two treatments are set, three treatments are carried out in parallel, and the stimulating agent of the invention and 1 commercially available decomposed inoculant (high-efficiency strains such as bacillus, trichoderma viride and saccharomycetes) are added (the adding proportion is 5 percent of the mass of the composting raw materials). And (5) comparing the rotten effect. 6 equal portions of the starting material are placed in 6 incubators 1050X 550X 500 mm. The compost box is internally provided with intermittent oxygen to ensure an aerobic fermentation process. The temperature of the contents was measured every 24 hours. Samples (about 200g) were collected from 5 different locations of each vessel during different temperatures for analysis, with the samples located about 25cm below the surface. After each sampling, the five samples were thoroughly mixed to form a homogeneous mixed sample.
Making a temperature change curve, as shown in figure 1, and the result shows that under the same condition, the treatment of adding the excitation preparation of the invention is advanced by 4 days and enters a high temperature period compared with the treatment of a commercial decomposition agent; and the high temperature period lasts long.
The method adopts a real-time fluorescent quantitative PCR technology to analyze the 16s rDNA of the compost sample, and comprises the following specific steps:
(1) extracting total DNA of organic materials: total DNA of the digested organic materials Using the Power MaxSoil DNA Isolation Kit from MOBIO, USA, 0.25g of each sample was weighed and total DNA of soil was extracted according to the Kit instructions.
(2) The 16S rRNA gene was determined using SYBR Green quantitative PCR method and the reaction was performed on an ABI 7500 Real-time PCR (ABI, USA) instrument.
RealTime PCR sample detection
1. The operation process is as follows:
1) and preparing a Realtime PCR reaction system for all the DNA samples. The system is configured as follows:
the solution was mixed by flicking the bottom of the tube and centrifuged briefly at 5000 rpm.
2) Sample application
a. 18ul of the mixture was added to each well corresponding to the 96-PCR plate.
b. The corresponding 2. mu.l of DNA was added.
c. Sealing Film Sealing Film was carefully glued on and briefly mixed by centrifugation.
d. The prepared PCR plate was placed on ice before setting up the PCR program.
3) Standard product configuration
3.1PCR amplification
Using the sample provided by the client as a template, and carrying out PCR amplification by using the target gene primer. The reaction system is shown in table 1:
TABLE 1 reaction System
| Composition (I) | Volume (μ l) |
| Target gene upstream primer (10. mu.M) | 1 |
| Target gene downstream primer (10. mu.M) | 1 |
| No. 1 template | 1 |
| 2× |
25 |
| Water (W) | 22 |
| In |
50 |
The amplification condition is 30 cycles of 94 ℃ 30S, 55 ℃ 30S and 72 ℃ 30S after pre-denaturation at 94 ℃ for 5min, and extension is carried out at 72 ℃ for 10min after the last 1 cycle is completed. After the reaction is finished, the amplification result is checked by using 1% agarose gel electrophoresis, the target fragment is recovered by agarose gel, and the steps are shown in the instructions of the agarose gel DNA recovery kit in century.
3.2 cloning of TA
The recovered PCR products were subjected to TA ligation, and the ligation system is shown in Table 2:
TABLE 2 connection System
| Composition (I) | Volume (μ l) |
| Recovered target fragments | 4 |
| T vector | 1 |
| 2× |
5 |
| In |
10 |
The cells were ligated at 22 ℃ for about 4 hours, and transformed into competent cells DH5 alpha, according to the protocol for use with DH5 alpha competent cells of the well-known century.
3.3 colony PCR identification of Positive clones
White colonies were picked, suspended in 10. mu.l of sterile water, and 1. mu.l was used as a template for colony PCR, and the rest was stored at 4 ℃ in a colony PCR system shown in Table 3:
TABLE 3 colony PCR System
| Composition (I) | Volume (μ l) |
| Target gene upstream primer (10. mu.M) | 0.2 |
| Target gene downstream primer (10. mu.M) | 0.2 |
| Form panel | 1 |
| 2× |
5 |
| Water (W) | 3.6 |
| In |
10 |
The amplification condition is 30 cycles of 94 ℃ 30S, 55 ℃ 30S and 72 ℃ 30S after pre-denaturation at 94 ℃ for 5min, and extension is carried out at 72 ℃ for 10min after the last 1 cycle is completed.
After completion of the reaction, the amplification result was checked by electrophoresis on 1% agarose gel.
3.4 extraction of plasmids
Plasmids were extracted as absolute quantitative standards.
The plasmid standard is 101-105And (3) performing 10-fold gradient dilution, and taking 2ul of each gradient as a template to establish a standard curve.
4) And placing the 96-PCR plate on a Realtime PCR instrument for PCR reaction.
The following procedure was followed: at 95 ℃ for 30 seconds; 40 PCR cycles (95 ℃, 5 sec; 60 ℃, 40 sec (fluorescence collection)). In order to establish the melting curve of the PCR product, after the amplification reaction is finished, the temperature is controlled according to the formula (95 ℃, 10 seconds, 60 ℃, 60 seconds, 95 ℃, 15 seconds); and slowly heated from 60 ℃ to 99 ℃ (instrument auto-run-Ramp Rate 0.05 ℃/sec).
The results of the experiment are shown in table 4. Two composting systems of excitant and commercial decomposing inoculantThere is a significant difference in the trend of the gene abundance change of the system. In the fertilizer pile added with the stimulating preparation of the invention, the copy number of 16s rDNA is 2.37 multiplied by 1011～ 8.68×1011Per gram of sample. The copy number of 16s rDNA per gram of sample was 1.36X 10 under the action of a commercial decomposition agent10～ 1.58×1011. The result shows that the bacteria generated in the running process of the compost of the exciting preparation are more, and the decomposition speed of the organic matters is faster than that of the compost of the commercial decomposing inoculant.
TABLE 4 copy number of 16s rDNA during different temperature periods for two composting systems, booster and commercial composter
| Treatment of | Commercial Maturity agent/average (/ 10)10) | Challenge formulation/mean (/ 10)11) |
| Period of temperature rise | 8.73 | 8.28 |
| High temperature period | 15.86 | 5.39 |
| Period of temperature reduction | 7.41 | 8.67 |
| Rotten period | 1.36 | 2.37 |
Example 2 preparation of priming formulations
Activating bacillus subtilis, and culturing in a seed culture medium: the broth culture medium comprises the following components: 10.0g of beef extract, 10.0g of peptone, 10.0g of glucose, 5.0g of NaCl, 1.0L of distilled water and pH 7.0.
Trichoderma harzianum activation, seed culture medium: the PDA culture medium has the following formula: 1.0L of potato extract, 20.0g of glucose and natural pH. Wherein, the potato extract: removing peel of potato 200g, cutting into small pieces, adding water 1.0L, boiling for 30min, filtering to remove potato pieces, and adding filtrate to 1.0L.
The formula of the composite fermentation medium comprises: 1.5% of corn flour, 0.5% of corn steep liquor, 5% of corn straw powder, 1.5% of starch, 2% of medium-temperature soybean cake powder, 0.6% of fish meal, and FeSO4 0.1％，MnSO4 0.1％，KH2PO4 0.1％，MgSO4 0.05％， CuSO40.05%, make up the filtrate to 1.0L, and adjust the pH to 7.0.
First, preparation method
1. Preparation of the liquid component a packaged separately:
b, bacillus subtilis seed liquid:
activating strains: culturing the bacillus subtilis strain in an activation culture medium for later use;
seed liquid: oscillating the activated bacillus subtilis strain at 37 ℃ for 12h to obtain a bacillus subtilis seed solution after culture;
trichoderma harzianum seed liquid:
activating strains: activating trichoderma harzianum for later use;
seed liquid: shaking the activated trichoderma harzianum strain at 30 ℃ for 24h, and culturing to obtain trichoderma harzianum seed liquid;
mixing zymophyte liquid: performing high-temperature sterilization on a composite fermentation culture medium, then respectively inoculating a bacillus subtilis seed solution and a trichoderma harzianum seed solution according to the inoculation amount of 5% when the temperature is reduced to be below 30 ℃, performing fermentation culture for 24 hours under the conditions that the temperature is 32 ℃ and the stirring speed is 180rpm, and performing fermentation culture for 60 hours under the conditions that the temperature is 28 ℃ and the stirring speed is 150 ℃; after the fermentation culture is finished, concentrating the fermentation liquor until the total effective viable count is more than or equal to 1000 hundred million CFU/g and the ratio of the bacillus subtilis to the trichoderma harzianum is more than or equal to 10:1, thus obtaining the mixed fermentation bacterial liquid.
2. Preparation of separately packaged solid component B:
mixing the following components in parts by weight: 500 parts of bacillus megatherium solid microbial inoculum, 400 parts of biochemical fulvic acid raw powder, 300 parts of charcoal, 0.01 part of gibberellic acid crystal powder and 0.0001 part of indoleacetic acid.
Second, formula
This example elicited formulation comprised an individually packaged liquid component a and an individually packaged solid component B;
the mass ratio of the liquid component A packaged independently to the solid component B packaged independently is 1: 3;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum; the total effective viable count of the mixed fermentation bacteria liquid of the bacillus subtilis and the trichoderma harzianum is 1000 hundred million CFU/g, wherein the ratio of the bacillus subtilis to the trichoderma harzianum is 10: 1;
the independently packaged component B consists of the following components in parts by weight:
500 parts of bacillus megatherium solid microbial inoculum, 400 parts of biochemical fulvic acid raw powder, 300 parts of charcoal, 0.01 part of gibberellic acid crystal powder and 0.0001 part of indoleacetic acid.
Third, the effect of use
The composting test is carried out by taking garden waste as a main raw material. The garden waste is obtained from agriculture and forestry academy of sciences of Beijing to make green pruning of branches and fallen leaves; the kitchen was collected from the worker canteen of the academy of agriculture and forestry, Beijing. The dried branches are crushed and cut into pieces with the length of 2-4 cm. The mass ratio of the raw materials is garden waste: and (3) when the kitchen is 7:3, controlling the C/N of the mixed material to be 25-35.
When the composting is started, the humidity is adjusted to about 60 percent. Two treatments are set, three treatments are carried out in parallel, and the stimulating preparation of the invention and 1 commercially available decomposed microbial inoculum (bacillus subtilis, bacillus licheniformis, lactobacillus and the like) are added (the adding proportion is 3 percent of the mass of the composting raw materials). And (5) comparing the rotten effect. 6 equal portions of the starting material are placed in 6 incubators 1050X 550X 500 mm. The compost box is internally provided with intermittent oxygen to ensure an aerobic fermentation process. The temperature of the contents was measured every 24 hours. Samples (about 200g) were collected from 5 different locations of each vessel during different temperatures for analysis, with the samples located about 25cm below the surface. After each sampling, the five samples were thoroughly mixed to form a homogeneous mixed sample.
Making a temperature change curve, as shown in fig. 2, and the result shows that under the same condition, the treatment of adding the excitation preparation of the invention is advanced by 4 days compared with the treatment of a commercial decomposition agent and enters a high temperature period; and the high temperature period lasts long.
The 16s rDNA of the compost sample is analyzed by real-time fluorescent quantitative PCR technology, and the specific steps are the same as example 1.
The results of the experiment are shown in Table 5. The gene abundance change trends of the two composting systems of the excitant preparation and the commercial decomposing inoculant are remarkably different. In the fertilizer pile added with the stimulating preparation of the invention, the copy number of 16s rDNA is 1.07 multiplied by 10 11～ 4.38×1011Per gram of sample. The copy number of 16s rDNA per gram of sample was 1.56X 10 under the action of a commercial decomposition agent10～ 1.33×1011. The result shows that the bacteria generated in the running process of the compost of the exciting preparation are more, and the decomposition speed of the organic matters is faster than that of the compost of the commercial decomposing inoculant.
TABLE 5 copy number of 16s rDNA during different temperature periods for two composting systems, spiked formulation and commercial decomposing agent
| Treatment of | Commercial Maturity agent/average (/ 10)10) | Challenge formulation/mean (/ 10)11) |
| Period of temperature rise | 5.73 | 4.38 |
| High temperature period | 13.32 | 2.35 |
| Period of temperature reduction | 4.41 | 4.27 |
| Rotten period | 1.56 | 1.07 |
Claims (10)
1. An organic waste compost stimulating agent, which is characterized in that: comprises an independently packaged liquid component A and an independently packaged solid component B;
the independently packaged liquid component A is a mixed fermentation bacterial liquid of bacillus subtilis and trichoderma harzianum;
the independently packaged solid component B consists of the following components in parts by weight: 300-600 parts of phosphate-solubilizing bacteria agent, 200-600 parts of biochemical fulvic acid raw powder, 300-600 parts of biochar, 0.005-0.015 part of gibberellic acid solid preparation and 0.00001-0.01 part of auxin.
2. The organic waste compost stimulating formulation of claim 1, wherein: the mass ratio of the independently packaged liquid component A to the independently packaged solid component B is 1: (1.5-4).
3. An organic waste compost stimulating formulation as claimed in claim 1 or claim 2 wherein: the total effective viable count of the mixed fermentation bacteria liquid of the bacillus subtilis and the trichoderma harzianum is more than or equal to 1000 hundred million CFU/g; and/or the presence of a gas in the gas,
in the mixed fermentation bacterial liquid of the bacillus subtilis and the trichoderma harzianum, the ratio of the bacillus subtilis to the trichoderma harzianum is more than or equal to 10: 1.
4. an organic waste compost stimulating formulation as claimed in any of claims 1 to 3 wherein: the mixed fermentation bacterial liquid of the bacillus subtilis and the trichoderma harzianum is prepared by the following steps: inoculating the seed solution of the bacillus subtilis and the seed solution of the trichoderma harzianum into a composite fermentation culture medium, and finishing fermentation culture;
the composite fermentation medium substrate is prepared by the following steps: mixing the following components with the balance of water according to the mass percentage of 100 percent: 1.5% of corn flour, 0.5% of corn steep liquor, 5% of corn straw powder, 1.5% of starch, 2% of medium-temperature soybean cake powder, 0.6% of fish meal, and FeSO4 0.1％，MnSO4 0.1％，KH2PO4 0.1％，MgSO40.05％，CuSO40.05 percent to obtain a mixed solution; regulating and controlling the pH value of the mixed solution to 7.0 to obtain the composite fermentation medium substrate.
5. The organic waste compost stimulating formulation of claim 4, wherein: the inoculation amount of the seed liquid of the bacillus subtilis and the inoculation amount of the seed liquid of the trichoderma harzianum are both 5-10%; and/or the presence of a gas in the gas,
the fermentation culture steps are as follows: firstly, fermenting and culturing for 24-48 h under the conditions that the temperature is 32-35 ℃ and the stirring speed is 150-180 rpm; and then fermenting and culturing for 48-72 h under the conditions that the temperature is 25-28 ℃ and the stirring speed is 120-150 rpm.
6. An organic waste compost stimulating formulation as claimed in any of claims 1 to 5 wherein: the phosphate solubilizing bacterium agent is a bacillus megaterium solid agent; and/or the presence of a gas in the gas,
the number of effective viable bacteria in the phosphate solubilizing bacteria agent is more than or equal to 100 hundred million CFU/g.
7. An organic waste compost stimulating formulation as claimed in any of claims 1 to 6 wherein: in the biochemical fulvic acid raw powder, the mass percentage content of biochemical fulvic acid is more than or equal to 90%; and/or the presence of a gas in the gas,
in the gibberellic acid solid preparation, the mass percentage of the gibberellic acid is 75%; and/or the presence of a gas in the gas,
the auxin is indoleacetic acid.
8. Use of an organic waste compost challenge formulation as claimed in any one of claims 1 to 7 in the manufacture of a product for stimulating rapid decomposition of organic waste.
9. A method of stimulating rapid decomposition of organic waste using a stimulating formulation as defined in any one of claims 1 to 7, comprising the steps of: and composting the organic waste added with the exciting agent, and quickly decomposing the organic waste after composting.
10. The method for stimulating rapid decomposition of organic waste according to claim 9, wherein: the adding amount of the excitation preparation is 1-5% of the mass of the organic waste.
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