Initiating compound microbial agent for activating aerobic compost carbon nitrogen emission reduction and application thereof

Abstract

The invention discloses a detonating compound microbial agent for activating aerobic composting carbon and nitrogen emission reduction and application thereof, and belongs to the technical field of microbial agent preparation. The preparation method comprises the following steps: screening bacillus subtilis, pseudomonas aeruginosa and gamma-aminobutyric acid to form a compost detonating agent, using chlorella, spirulina platensis and staphylococcus pasteurii to form a phycomycete symbiont as an auxiliary agent, and using a mixture of corncob powder and zeolite powder as a strain carrier; the prepared detonating compound microbial agent for activating aerobic composting carbon nitrogen emission reduction has the advantages of promoting carbon nitrogen emission reduction, being small in addition amount, high in composting efficiency and high in environment adaptability, is suitable for fermentation of organic matter such as crop straw and livestock and poultry manure, is wide in application range and has wide market application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial agent preparation, and in particular relates to an explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting and an application thereof. Background Art

[0002] Aerobic composting is the process by which microorganisms decompose and stabilize organic matter in materials under aerobic conditions to produce decomposed fertilizer. As an effective way to recycle waste resources, aerobic composting not only produces large quantities of nitrogen, phosphorus, and potassium compounds that can be absorbed by plants, but also synthesizes humus, an important active organic matter that improves soil fertility.

[0003] The traditional aerobic composting process is a physiological and biochemical process involving the natural microbial community in the compost. Although composting crop straw, livestock and poultry manure, and kitchen waste is an ideal way to recycle waste organic matter, many problems still exist. These include insufficient degradation of macromolecular organic matter, large amounts of greenhouse gas emissions during the composting process, and low maturity. Among them, greenhouse gases produced during the composting process mainly come from the decomposition of organic matter. The process of microbial decomposition of organic matter is also the main process for releasing carbon dioxide (CO2); the degradation, nitrification, and denitrification of organic nitrogen will produce nitrous oxide (N2O). Currently, there are many studies on microbial agents that promote aerobic composting, but there are also many shortcomings. For example, by adding exogenous microorganisms, the number of microbial communities in the initial stage of composting can be increased and the community structure can be improved, thereby accelerating the decomposition of organic matter, accelerating the increase in pile temperature, extending the high temperature period, and shortening the composting time. However, many bacterial agents currently on the market have a single strain or microalgae technology and poor adaptability to the composting environment, which makes the microbial strains in them unable to play a role in the composting process or even become ineffective, and they cannot effectively inhibit the emissions of carbon dioxide (CO2), nitrous oxide (N2O) and ammonia (NH3) produced during the composting process.

[0004] At present, how to develop an efficient bacterial agent that can promote carbon and nitrogen emission reduction, has a good enzymatic decomposition effect on mainstream organic matter in aerobic composting, such as crop straw, livestock and poultry manure, and kitchen waste, and at the same time has a good inhibitory effect on greenhouse gas emissions generated during aerobic composting, is a technical problem that needs to be solved urgently. Summary of the Invention

[0005] To address the above technical issues, the present invention proposes an explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting, and its application. This explosive composite microbial agent can be added to specific composting processes, promoting rapid temperature rise in aerobic composting and accelerating the decomposition of organic matter while effectively reducing emissions of carbon dioxide (CO2), nitrous oxide (N2O), and ammonia (NH3) during aerobic composting. Suitable for the fermentation of organic matter such as straw and livestock and poultry manure, this product is suitable for large-scale applications.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] Technical Solution 1: An explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting, comprising the following raw materials: a compost initiator, an adjuvant, and an agent carrier; wherein,

[0008] The compost detonator consists of Bacillus subtilis, Pseudomonas aeruginosa and gamma-aminobutyric acid;

[0009] The auxiliary agent is composed of Chlorella vulgaris, Spirulina platensis and Staphylococcus pasteurianus;

[0010] The bacterial agent carrier is a mixture of corn cob powder and zeolite powder.

[0011] The present invention screens Bacillus subtilis, Pseudomonas aeruginosa and gamma-aminobutyric acid to form a compost initiator, uses Chlorella vulgaris, Spirulina platensis and Staphylococcus pasteurianus to form an algae-bacteria symbiont as an auxiliary agent, and accelerates the activation of microbial activity through the joint action of multiple algae and bacteria, effectively promoting the decomposition of organic matter, shortening the aerobic composting time, promoting the maturity of aerobic composting, improving the quality of compost products, and reducing the emissions of carbon dioxide (CO2), nitrous oxide (N2O) and ammonia (NH3) during the composting process. It not only improves the resource utilization efficiency of organic waste, but also promotes the virtuous cycle of the ecological environment. This method provides strong support for the scientific management of composting and the sustainable development of agricultural clean production.

[0012] Furthermore, the mass ratio of the Bacillus subtilis, Pseudomonas aeruginosa, and γ-aminobutyric acid is 2:(1.1-1.5):1.

[0013] The high-intensity metabolic activity of Bacillus subtilis and Pseudomonas aeruginosa during the composting process will generate a large amount of heat energy to quickly heat up the compost. At the same time, they secrete a variety of enzymes with strong stress resistance and good antibacterial effect, which decompose macromolecular organic matter such as protein and cellulose and convert them into small molecular compounds such as amino acids and sugars and inorganic salts. These products are not only easy to be further utilized by other microorganisms, but also increase the nutrient content and availability of the compost and promote the composting process. γ-aminobutyric acid acts as a signal molecule to activate the microorganisms in the compost, promote the growth and reproduction of microorganisms, accelerate the decomposition and conversion of organic matter in the composting process, regulate the metabolic activity of microorganisms, and improve the fertilizer efficiency of the compost. The present invention selects Bacillus subtilis, Pseudomonas aeruginosa and γ-aminobutyric acid to form a compost initiator. The three components can promote a significant increase in the compost fermentation rate through synergistic action, which is conducive to more complete fermentation and a shortened composting cycle.

[0014] Furthermore, the effective viable counts of Bacillus subtilis and Pseudomonas aeruginosa were both 1×10 10 cfu / mL.

[0015] Furthermore, the mass ratio of Chlorella, Spirulina platensis and Staphylococcus pasteurianus is (4-8):3:(1-5).

[0016] Chlorella and Spirulina platensis utilize the inorganic salts and carbon dioxide (CO2) in compost to reproduce and produce a variety of enzymes, such as cellulases and proteases. These enzymes accelerate the decomposition of organic matter, thereby promoting compost maturation. They also effectively fix CO2, reducing carbon emissions during the composting process. Furthermore, their ability to absorb and convert nitrogen in compost helps reduce nitrous oxide (N2O) and ammonia (NH3) emissions. They also promote the growth and reproduction of mesophilic and thermophilic microorganisms during the composting process, further accelerating compost maturation and improving soil fertility. Staphylococcus pasteurianus decomposes complex organic matter, such as cellulose and lignin, in aerobic composting, converting them into substances more easily absorbed by plants. The beneficial substances produced by their metabolic activities, such as antibiotics and enzymes, inhibit the growth of harmful microorganisms, thereby accelerating compost maturation and improving the fertilizer efficiency and hygienic quality of the compost.

[0017] Furthermore, the mass ratio of the corncob powder to the zeolite powder is 0.6:(1.1-1.5).

[0018] Technical Solution 2: Application of an explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting in the composting field.

[0019] Technical solution three: A composting method comprising the following steps:

[0020] The compost initiator and the microbial agent carrier in the initiating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic compost are mixed and granulated to obtain solid particles;

[0021] After adjusting the moisture content of the compost raw materials to 55-65%, the solid particles are added for compost fermentation. When the compost temperature rises to 45-50° C., the auxiliary agent in the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting is sprayed onto the surface of the pile, and the fermentation is continued until the compost is completed.

[0022] The present invention adds algae-fungus symbionts during the composting process. As an auxiliary agent, the algae-fungus symbiont can significantly promote the rapid decomposition and conversion of organic matter, improve the efficiency and quality of composting, and more importantly, effectively reduce the emission of greenhouse gases such as carbon dioxide (CO2) and nitrous oxide (N2O) by optimizing microbial activity, and significantly inhibit the volatilization of ammonia (NH3), thereby achieving the dual environmental benefits of carbon and nitrogen emission reduction and ammonia inhibition.

[0023] Furthermore, the mass ratio of the compost initiator to the bacterial agent carrier is (4-8): (16-20).

[0024] Furthermore, the amount of the solid particles is 1.5-2.5‰ of the mass of the compost raw materials; and / or

[0025] The amount of the auxiliary agent is 1.5-2.5‰ of the mass of the compost raw material; and / or

[0026] The composting fermentation refers to fermenting at a temperature of 10-30° C. until the composting is completed.

[0027] Compared with the prior art, the present invention has the following advantages and technical effects:

[0028] This invention has the advantages of promoting carbon and nitrogen emission reduction, requiring minimal additives, achieving high composting efficiency, and strong environmental adaptability. It is suitable for the fermentation of organic matter such as crop straw and livestock and poultry manure, and has a wide range of applications and promising market prospects. It represents a positive response to promoting the resource utilization of agricultural waste, reducing carbon and nitrogen emissions in the agricultural sector, promoting green and clean production, and achieving sustainable agricultural development. DETAILED DESCRIPTION

[0029] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0030] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0032] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative only.

[0033] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0034] The present invention provides an explosive composite microbial inoculant for activating carbon and nitrogen emission reduction in aerobic composting. The inoculant comprises the following ingredients: a compost inoculant, an additive, and an inoculant carrier. This inoculant effectively activates microbial activity during aerobic composting, improving the decomposition efficiency of organic matter. Furthermore, through the action of algae-bacteria symbionts, it promotes carbon and nitrogen cycling, reduces greenhouse gas and ammonia emissions, and thus enhances the quality and environmental benefits of the compost product.

[0035] In some embodiments of the present invention, the compost initiator consists of Bacillus subtilis, Pseudomonas aeruginosa and γ-aminobutyric acid.

[0036] In some embodiments of the present invention, the method for preparing the compost initiator comprises the following steps: activating and expanding Bacillus subtilis and Pseudomonas aeruginosa separately to obtain an effective viable cell count of 1×10 10cfu / mL of bacterial liquid, and then Bacillus subtilis, Pseudomonas aeruginosa, and γ-aminobutyric acid are mixed in a mass ratio of 2: (1.1-1.5): 1. Exemplarily, in some embodiments of the present invention, Bacillus subtilis, Pseudomonas aeruginosa, and γ-aminobutyric acid are compounded in a mass ratio of 2: 1.1: 1, 2: 1.2: 1, 2: 1.3: 1, 2: 1.4: 1 or 2: 1.5: 1.

[0037] In some embodiments of the present invention, the auxiliary agent is composed of Chlorella, Spirulina platensis and Staphylococcus pasteurianus in a mass ratio of (4-8):3:(1-5). Exemplarily, Chlorella, Spirulina platensis and Staphylococcus pasteurianus are compounded in a mass ratio of 4:3:1, 5:3:2, 6:3:3, 7:3:4 or 8:3:5 to form the auxiliary agent; the Staphylococcus pasteurianus in the auxiliary agent is deposited with CGMCC No. 13423, and other raw materials are commercially available.

[0038] In some embodiments of the present invention, the bacterial agent carrier is formed by mixing corn cob powder and zeolite powder in a mass ratio of 0.6:(1.1-1.5). Exemplarily, the corn cob powder and zeolite powder are compounded in a mass ratio of 0.6:1.1, 0.6:1.2, 0.6:1.3, 0.6:1.4 or 0.6:1.5.

[0039] In some embodiments of the present invention, the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting can be widely used in the field of composting, such as the fermentation of organic matter such as crop straw and livestock and poultry manure.

[0040] An embodiment of the present invention further provides a composting method, comprising the following steps:

[0041] (1) The compost detonator and the agent carrier in the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic compost are mixed and granulated to obtain solid particles with a particle size of 2-4 mm; 2-4 mm is acceptable, and for example, solid particles with a particle size of 2 mm are selected in the following examples of the present invention;

[0042] (2) After adjusting the moisture content of the compost raw materials to 55-65%, the solid particles are added to carry out composting fermentation. When the compost temperature rises to 45-50° C., the auxiliary agent in the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting is sprayed onto the surface of the pile body, and the fermentation is continued until the composting is completed.

[0043] In some preferred embodiments, in step (1), the mass ratio of the compost initiator to the microbial agent carrier is (4-8):(16-20). Exemplarily, the compost initiator and the microbial agent carrier are compounded in a mass ratio of 4:16, 5:17, 6:18, 7:19 or 8:20.

[0044] In some preferred embodiments, in step (1), a water-activated epoxy resin may be added during granulation to promote granulation. The specific granulation method may be a conventional method in the art, and the specific process is not considered as a factor in the inventiveness of the present invention.

[0045] In some preferred embodiments, in step (1), the amount of the solid particles is 1.5-2.5‰ of the mass of the compost raw material. For example, in the following embodiments of the present invention, the effect is discussed by taking the amount of the solid particles as 1.8‰ of the mass of the compost raw material as an example.

[0046] In some preferred embodiments, in step (2), the amount of the auxiliary agent is 1.5-2.5‰ of the mass of the compost raw material. For example, in the following embodiments of the present invention, the effect is discussed by taking the amount of the auxiliary agent as 2.2‰ of the mass of the compost raw material as an example.

[0047] In some preferred embodiments, in step (2), the composting fermentation refers to fermentation at a temperature of 10-30° C. until the composting is completed. For example, in the following embodiments of the present invention, the composting fermentation refers to fermentation at a temperature of 26° C. until the composting is completed.

[0048] In some preferred embodiments, in step (2), the moisture content of the compost raw material is adjusted to 55-65% when the compost raw material is first adjusted. For example, the following embodiments of the present invention take a moisture content of 55% as an example to discuss the effect.

[0049] Unless otherwise specified, the "room temperature" in the present invention refers to 20-30°C.

[0050] The raw materials used in the present invention are all purchased from the market.

[0051] The technical solution of the present invention is further illustrated by the following examples.

[0052] In the following embodiments of the present invention, in actual application, organic materials (the organic raw materials are corn stalks and cow dung, and the preparation method is: crushing the corn stalks to a particle size of 3-6 cm and mixing them with cow dung. The mixing ratio is arbitrary, and the proportion is kept uniform in the following embodiments) are humidified, adjusted to 55%, and piled into a pile of length×width×height=1.5m×1.2m×1.5m. Then, solid particles and additives in the detonating composite microbial inoculant for activating carbon and nitrogen emission reduction in aerobic composting are respectively sprinkled according to the composting period for aerobic composting. When the carbon-nitrogen ratio is (20-22): 1, the pH is 7.5-8, and the germination index is greater than 95%, it indicates that the composting is completely mature, that is, the composting is completed.

[0053] The germination index (GI) of compost is an important indicator for evaluating compost maturity, primarily used to assess the toxic effects of compost on plant growth. The GI measures the germination rate and root length of seeds in compost extract to reflect the phytotoxicity of compost. Specifically, a higher GI indicates a compost with less toxicity to plant growth and a higher degree of maturity.

[0054] In the following examples, Bacillus subtilis was purchased from the China General Microorganism Culture Collection Center with accession number CGMCC1.14985; Pseudomonas aeruginosa was purchased from the China General Microorganism Culture Collection Center with accession number CGMCC1.10712; and γ-aminobutyric acid was purchased from Zhengzhou Chaofan Chemical Co., Ltd. The slant culture medium used for Bacillus subtilis was tryptone soy agar, cultured at 30°C for 2-3 days; the slant culture medium used for Pseudomonas aeruginosa was nutrient broth agar, cultured at 37°C for 1-2 days; and the liquid culture medium used for Bacillus subtilis consisted of 1% tryptone, 0.5% yeast extract, and 1% sodium chloride, with the pH adjusted to 7.2-7.4 with 2 mol / L sodium hydroxide. The liquid culture medium used for Pseudomonas aeruginosa consists of 3.0% beef extract, 5% peptone, 5.0% NaCl, and 0.005% MnSO₄·H₂O to promote spore formation. The pH is adjusted to 7.0-7.2 with 1 mol / L sodium hydroxide. Other parameters and unspecified operating procedures in the culture methods for various microorganisms can be referenced by conventional methods in the art, and the specific details are not considered as factors contributing to the innovative nature of the present invention.

[0055] In the following examples of the present invention, the preparation method of the auxiliary agent includes the following steps: culturing Chlorella, Spirulina platensis and Staphylococcus pasteurianus in a solid culture medium to the logarithmic phase, picking up single colonies of Chlorella, Spirulina platensis and Staphylococcus pasteurianus and inoculating them into TAP culture medium, culturing them under light in a shaking table, discarding the supernatant, and continuing light culturing to obtain the algae-bacteria symbiont. The light intensity of the shaking table light culture is 120 μmol / m 2 ·s, the light source was a white LED lamp, the culture temperature was 30°C, the shaker speed was 150 r / min; the light intensity of the light culture was 120 μmol / m 2 s, the light source is a white LED lamp, and the culture temperature is 30° C. Other parameters and methods not specifically limited in the preparation method can refer to conventional methods in the art, and the specific content is not considered as a factor in the innovative discussion of the present invention.

[0056] Example 1

[0057] S1. Preparation of compost initiator:

[0058] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×10 10 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.1:1 to obtain a compost initiator;

[0059] S2. Preparation of additives:

[0060] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 4:3:1 and cultured on a solid culture medium until the logarithmic phase. Single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into TAP culture medium, cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination for 72 hours to obtain the algae-bacteria symbiont, i.e., the adjuvant;

[0061] S3. Preparation of bacterial agent carrier

[0062] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200-mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.1;

[0063] S4. Preparation of solid particles

[0064] The compost initiator in S1 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 4:16, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0065] S5. Composting and fermentation

[0066] After adjusting the moisture content of the compost raw material (cow dung) to 55%, solid particles of S4 are added (the amount of solid particles added is 1.8‰ of the mass of the compost raw material), and composting is carried out at 26°C. When the compost temperature rises to between 45-50°C, the auxiliary agent in S2 is sprayed onto the surface of the pile (the amount of the auxiliary agent is 2.2‰ of the mass of the compost raw material), and fermentation is continued until the compost is completed.

[0067] Example 2

[0068] S1. Preparation of compost initiator:

[0069] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×10 10 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.2:1 to obtain a compost initiator;

[0070] S2. Preparation of additives:

[0071] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 5:3:2 and cultured on a solid culture medium until the logarithmic phase. Single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into a TAP culture medium. The mixture is cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination to obtain the algae-bacteria symbiont, i.e., the adjuvant.

[0072] S3. Preparation of bacterial agent carrier

[0073] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200-mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.2;

[0074] S4. Preparation of solid particles

[0075] The compost initiator in S1 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 5:17, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0076] S5. Composting and fermentation

[0077] After adjusting the moisture content of the compost raw materials to 55%, add the solid particles of S4 (the amount of solid particles added is 1.8‰ of the mass of the compost raw materials), and carry out composting fermentation at 26°C. When the compost temperature rises to between 45-50°C, spray the additives in S2 onto the surface of the pile (the amount of additives is 2.2‰ of the mass of the compost raw materials), and continue fermenting until the compost is completed.

[0078] Example 3

[0079] S1. Preparation of compost initiator:

[0080] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×10 10 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.3:1 to obtain a compost initiator;

[0081] S2. Preparation of additives:

[0082] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 6:3:3 and cultured on a solid culture medium until the logarithmic phase. Single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into a TAP culture medium. The mixture is cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination to obtain the algae-bacteria symbiont, i.e., the adjuvant.

[0083] S3. Preparation of bacterial agent carrier

[0084] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200-mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.3;

[0085] S4. Preparation of solid particles

[0086] The compost initiator in S1 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 6:18, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0087] S5. Composting and fermentation

[0088] After adjusting the moisture content of the compost raw materials to 55%, add the solid particles of S4 (the amount of solid particles added is 1.8‰ of the mass of the compost raw materials), and carry out composting fermentation at 26°C. When the compost temperature rises to between 45-50°C, spray the additives in S2 onto the surface of the pile (the amount of additives is 2.2‰ of the mass of the compost raw materials), and continue fermenting until the compost is completed.

[0089] Example 4

[0090] S1. Preparation of compost initiator:

[0091] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×10 10 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.4:1 to obtain a compost initiator;

[0092] S2. Preparation of additives:

[0093] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 7:3:4 and cultured on a solid culture medium until the logarithmic phase, and single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into a TAP culture medium, cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination to obtain the algae-bacteria symbiont, i.e., the adjuvant;

[0094] S3. Preparation of bacterial agent carrier

[0095] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200 mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.4;

[0096] S4. Preparation of solid particles

[0097] The compost initiator in S1 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 7:19, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0098] S5. Composting and fermentation

[0099] After adjusting the moisture content of the compost raw materials to 55%, add the solid particles of S4 (the amount of solid particles added is 1.8‰ of the mass of the compost raw materials), and carry out composting fermentation at 26°C. When the compost temperature rises to between 45-50°C, spray the additives in S2 onto the surface of the pile (the amount of additives is 2.2‰ of the mass of the compost raw materials), and continue fermenting until the compost is completed.

[0100] Example 5

[0101] S1. Preparation of compost initiator:

[0102] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×10 10 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.5:1 to obtain a compost initiator;

[0103] S2. Preparation of additives:

[0104] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 8:3:5 and cultured on a solid culture medium until the logarithmic phase. Single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into TAP culture medium, cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination to obtain the algae-bacteria symbiont, i.e., the adjuvant;

[0105] S3. Preparation of bacterial agent carrier

[0106] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200-mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.5;

[0107] S4. Preparation of solid particles

[0108] The compost initiator in S1 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 8:20, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0109] S5. Composting and fermentation

[0110] After adjusting the moisture content of the compost raw materials to 55%, add the solid particles of S4 (the amount of solid particles added is 1.8‰ of the mass of the compost raw materials), and carry out composting fermentation at 26°C. When the compost temperature rises to between 45-50°C, spray the additives in S2 onto the surface of the pile (the amount of additives is 2.2‰ of the mass of the compost raw materials), and continue fermenting until the compost is completed.

[0111] Comparative Example 1

[0112] The method is the same as Example 1, except that Chlorella is not added to the auxiliary agent of S2, and the mass ratio of Spirulina platensis to Staphylococcus pasteurianus is maintained at 3:1.

[0113] Comparative Example 2

[0114] The method is the same as Example 1, except that Spirulina platensis is not added to the auxiliary agent of S2, and the mass ratio of Chlorella and Staphylococcus pasteurianus is maintained at 4:1.

[0115] Comparative Example 3

[0116] The method is the same as Example 1, except that Staphylococcus pasteurianus is not added to the auxiliary agent of S2, and the mass ratio of Chlorella vulgaris to Spirulina platensis is maintained at 4:3.

[0117] Comparative Example 4

[0118] The same as Example 1, except that in S2, Chlorella, Spirulina platensis and Staphylococcus pasteurianus are mixed in a mass ratio of 9:4:6.

[0119] Comparative Example 5

[0120] The method is the same as Example 1, except that in S2, the mass ratio of Chlorella, Spirulina platensis, Staphylococcus pasteurianus and Streptomyces is 4:3:1:3.

[0121] Comparative Example 6

[0122] The method is the same as Example 1, except that in S1, Bacillus subtilis, Pseudomonas aeruginosa, and γ-aminobutyric acid are mixed in a mass ratio of 3:4:2.

[0123] Comparative Example 7

[0124] The method is the same as Example 1, except that in S1, Bacillus subtilis, Pseudomonas aeruginosa, γ-aminobutyric acid, and Bacillus circulans are mixed in a mass ratio of 2:1.1:1:2.

[0125] Comparative Example 8

[0126] The method is the same as Example 1, except that the compost initiator and the bacterial agent carrier in S4 are mixed in a mass ratio of 3:15.

[0127] Comparative Example 9

[0128] Preparation of an explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting:

[0129] S1. Preparation of compost initiator:

[0130] After Bacillus subtilis and Pseudomonas aeruginosa were activated and expanded separately, the effective viable count was 1×1010 cfu / mL of bacterial liquid, and then mixing the Bacillus subtilis bacterial liquid, the Pseudomonas aeruginosa bacterial liquid, and gamma-aminobutyric acid in a mass ratio of 2:1.1:1 to obtain a compost initiator;

[0131] S2. Preparation of additives:

[0132] Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are mixed in a mass ratio of 4:3:1 and cultured on a solid culture medium until the logarithmic phase, and single colonies of Chlorella, Spirulina platensis, and Staphylococcus pasteurianus are respectively picked and inoculated into TAP culture medium, cultured on a shaking table under illumination, the supernatant is discarded, and the culture is continued under illumination to obtain the algae-bacteria symbiont, i.e., the adjuvant;

[0133] S3. Preparation of bacterial agent carrier

[0134] The corn cob powder and zeolite powder were fully dried, crushed and passed through a 200-mesh sieve, and then mixed evenly in a mass ratio of 0.6:1.1;

[0135] S4. Preparation of solid particles

[0136] The compost initiator in S1, the adjuvant in S2 and the bacterial agent carrier in S3 were mixed evenly in a mass ratio of 4:8:16, and placed in a granulator for granulation to obtain solid particles with a particle size of 2 mm;

[0137] S5. Composting and fermentation

[0138] After adjusting the moisture content of the compost raw material (cow dung) to 55%, solid particles of S4 were added (the amount of solid particles added was 1.8‰ of the mass of the compost raw material), and composting was carried out at 26°C until the composting was completed.

[0139] Blank control group:

[0140] The same as Example 1, except that no explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting is added for composting fermentation. Specifically, the moisture content of the compost raw materials is adjusted to 55%, and the composting fermentation is carried out at 26° C. until the composting is completed.

[0141] Performance Testing(1)

[0142] During the aerobic composting process of the above examples and comparative examples, the maximum compost temperature, time to reach the maximum temperature, pH, carbon dioxide emission rate, nitrous oxide emission rate, ammonia emission rate, germination index, and composting period were measured to evaluate the effects of different auxiliary materials on cow dung composting. The above measurement indicators were tested according to conventional test methods. Among them, the germination index was determined as follows:

[0143] Compost samples were mixed with distilled water at a mass ratio of 1:15, shaken thoroughly, and then extracted in a 28°C incubator for 24 hours. Using a pipette, 5 mL of the filtrate was transferred to a 10 cm Petri dish lined with two 5 cm qualitative filter papers. Thirty grass seeds were sown in each dish and incubated in a 28°C incubator for 48 hours. The seed germination rate and root length were determined, and the germination index (GI) (%) was calculated. Each treatment was repeated three times. Seed germination rate is calculated using formula (1).

[0144]

[0145] The test results are shown in Table 1.

[0146] Table 1

[0147]

[0148] It can be seen from the data in Table 1 that the various indicators of the embodiments are generally better than those of the comparative examples. The maximum temperatures in Examples 1-5 are generally higher, and the time to reach the maximum temperature is relatively short, which is conducive to the microbial activity and rapid decomposition of organic matter during the composting process. At the same time, the pH values ​​of Examples 1-5 are maintained within an appropriate range, which is conducive to the growth and reproduction of microorganisms. This shows that in the present invention, each algae and fungus in the auxiliary agent plays an important role, and any deletion or adjustment has an impact on the overall effect of the composting. Therefore, the selection of the strain of the present invention is a key technical feature for achieving efficient composting and fermentation. Changing the type or dosage of the functional strain will lead to a decline in the fermentation effect. The three complement each other and are indispensable. It can also be seen that Example 1 is the best embodiment.

[0149] It can also be seen from the data in Table 1 that the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting prepared in Example 1 of the present invention is compared with the blank control group. Example 1 can not only improve the composting efficiency and shorten the cycle, but also optimize the compost quality, maintain a suitable pH, promote microbial activity and organic matter decomposition, thereby improving the aerobic fermentation efficiency of organic waste. More importantly, the additive prepared by the present invention can reduce the emissions of carbon dioxide (CO2), nitrous oxide (N2O) and ammonia (NH3) during the composting process. After applying the additive, the emission rate of CO2 is reduced from 124.7g / (dm 2 ) is reduced to 99.2g / (dm 2 ), the emission rate of N2O increased from 3.9g / (dm 2 ) is reduced to 1.2g / (dm 2 ), the emission rate of NH3 is from 27.6g / (dm 2 ) is reduced to 18.1g / (dm 2). It reduces the emissions of CO2, N2O and NH3, improves the efficiency of carbon-nitrogen conversion in composting, and reduces greenhouse gas and odor emissions during aerobic composting.

[0150] The present invention optimizes the aerobic composting process in a targeted manner by adding a compost initiator and an auxiliary agent during different aerobic composting processes, thereby promoting rapid temperature rise of the aerobic composting pile, with the maximum temperature reaching 67.8°C. The temperature of the pile is rapidly increased within 24 hours, and the seed germination rate reaches 131.2% at the end of composting. This indicates that the compost to which the compost initiator and the auxiliary agent are added has a positive effect on rapid temperature rise of the pile, increased compost temperature, sufficient decomposition of organic matter, and promotion of seed germination.

[0151] Performance Testing(2)

[0152] When growing tobacco indoors using a substrate, the problem of repeated cropping is serious because tobacco secretes a large amount of harmful small molecules that breed harmful microorganisms. Supplementing 5% sheep manure-based compost during tobacco cultivation can effectively retain nutrients such as low-molecular-weight organic carbon, ammonium nitrogen, and nitrate nitrogen due to the reduction in carbon and nitrogen emissions during the composting process, providing sufficient nutrients for tobacco growth. In addition, the beneficial microorganisms (Bacillus subtilis, Pseudomonas aeruginosa, Chlorella, Spirulina platensis, and Staphylococcus pasteurianus) and gamma-aminobutyric acid added during the composting process all promote tobacco growth. At the same time, the above-mentioned microorganisms can significantly inhibit the problem of repeated cropping caused by the activity of Fusarium and Phytophthora parasiticus.

[0153] The microbial compost was applied to the indoor substrate of tobacco in Neijiang, Sichuan (the mechanism is the soil for growing tobacco). The reused substrate had serious re-cropping due to the above reasons, and the tobacco seed germination rate and growth condition were poor. After the secondary substrate was initially inactivated by high-temperature dry heat, 5% of the compost product prepared according to the method of Example 1 was added (i.e., the compost product in Example 1 was mixed with the repeated substrate as a new substrate). Laboratory experiments verified that the growth of Nicotiana benthamiana in the treated old substrate was consistent with or even better than the new substrate. The three-party verification confirmed that after further inoculation of the engineered microorganisms into the Nicotiana benthamiana leaves for 5 days, the biomass of the single plant in the new substrate was 4-6g, the biomass of the single plant in the treatment group was 4-5.8g, the target protein content of the new substrate was about 58μg / g FW, and the target protein content in the treatment substrate was about 55gμg / g FW, which was determined to be no significant difference by the three-party institutions. This further proves that the composite bacterial agent is advantageous in improving the efficiency of organic fertilizers and inhibiting the problem of re-cropping.

[0154] The above are merely preferred embodiments of the present application, but the scope of protection of the present application is not limited thereto. Any changes or substitutions that can be easily conceived by a person skilled in the art within the technical scope disclosed in this application should be included in the scope of protection of the present application. Therefore, the scope of protection of the present application should be based on the scope of protection of the claims.

Claims

1. An explosive composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting, characterized in that: The raw materials include: compost initiator, auxiliary agent and bacterial agent carrier; among which, The compost detonator is composed of Bacillus subtilis, Pseudomonas aeruginosa and gamma-aminobutyric acid; The auxiliary agent is composed of chlorella, spirulina platensis and Staphylococcus pasteurianus; The bacterial agent carrier is a mixture of corn cob powder and zeolite powder.

2. The detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to claim 1, characterized in that: The mass ratio of the Bacillus subtilis, Pseudomonas aeruginosa and gamma-aminobutyric acid is 2:(1.1-1.5):

1.

3. The detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to claim 2, characterized in that: The effective viable counts of Bacillus subtilis and Pseudomonas aeruginosa are both 1×10 10 cfu / mL.

4. The detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to claim 1, characterized in that: The mass ratio of the chlorella, spirulina platensis and Staphylococcus pasteurianus is (4-8):3:(1-5).

5. The detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to claim 1, characterized in that: The mass ratio of the corncob powder to the zeolite powder is 0.6:(1.1-1.5).

6. Use of the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting as claimed in any one of claims 1 to 5 in the field of composting.

7. A composting method, characterized in that: The following steps are involved: The compost detonator and the agent carrier in the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to any one of claims 1 to 5 are mixed and granulated to obtain solid particles; After adjusting the moisture content of the compost raw materials to 55-65%, the solid particles are added for composting fermentation. When the compost temperature rises to 45-50° C., the auxiliary agent in the detonating composite microbial agent for activating carbon and nitrogen emission reduction in aerobic composting according to any one of claims 1-5 is sprayed onto the surface of the pile, and the fermentation is continued until the composting is completed.

8. The composting method according to claim 7, characterized in that: The mass ratio of the compost detonator to the bacterial agent carrier is (4-8): (16-20).

9. The composting method according to claim 7, characterized in that: The amount of the solid particles is 1.5-2.5‰ of the mass of the compost raw material; and / or The dosage of the auxiliary agent is 1.5-2.5‰ of the mass of the compost raw material.

10. The composting method according to claim 7, characterized in that: The composting fermentation process is to ferment at a temperature of 10-30° C. until the composting is completed.

Images