Composite microbial agent for high-temperature composting and preparation method thereof

Abstract

The invention belongs to the field of microbial technology application, and particularly relates to a compound microbial agent for high-temperature composting and a preparation method thereof. According to the invention, a plurality of thermophilic fungi with different enzyme production functions and a plurality of thermophilic bacilli are compounded, and the preparation method adopts a manner of independently preparing the bacilli and the thermophilic fungi and then physically mixing the bacilli and the thermophilic fungi, so that the mutual influence is reduced, and the concentration and activity of the two types of microorganisms are guaranteed. In terms of technical effects, the microbial inoculum disclosed by the invention can be rapidly heated, the high-temperature period is longer, and a round of fermentation can be completed in a shorter time period. Experimental data show that the microbial inoculum prepared by the method can reach the high temperature of 75 DEG C within 2 days and can be kept in a high-temperature state to the tenth day, the fermentation period is short, one round of fermentation only needs 16 days, and the high-temperature time period can occupy about 56%-62.5% of the whole fermentation period.

Description

Technical Field

[0001] The invention belongs to the field of microbial technology application, and specifically relates to a composite microbial agent for high-temperature composting and a preparation method thereof. Background Art

[0002] Aerobic high-temperature composting is an effective method for treating urban feces today. Its principle is mainly the process of aerobic bacteria absorbing, oxidizing and decomposing feces under good ventilation and sufficient oxygen conditions. Aerobic microorganisms oxidize part of the absorbed organic matter into simple inorganic matter through their own life activities, while releasing energy required for microbial growth activities, while the other part of the organic matter is synthesized into new cytoplasm, allowing the microorganisms to continue to grow and reproduce, producing more organisms.

[0003] Modern composting processes usually include four stages: heating, high temperature, cooling, and post-ripening. In the past, composting temperatures above 50°C were usually called high temperature stages, but modern composting processes usually define high temperature stages as temperatures above 60°C. Usually, high temperatures above 65°C can kill pathogens, parasite eggs, and weed seeds, and achieve harmless treatment of organic waste.

[0004] Organic solid waste represented by urban feces has the characteristics of dense structure and high moisture content. It requires a large amount of plant-based auxiliary materials to be added. In the composting process, there are problems such as slow temperature rise and short high temperature period. Since high-quality compost has very strict temperature requirements, and most of them require higher temperatures, the existing technology solves this problem through a variety of different methods. For example, CN117844716A provides a microbial agent suitable for improving the conversion of compost organic matter under low temperature conditions and its preparation method, which avoids the problem of insufficient temperature by selecting strains adapted to low-temperature fermentation; for example, CN101486969A uses three strains of Candida utilis, Lactobacillus plantarum, and Bacillus subtilis, which are introduced in the early stage of composting. Under the promotion of exogenous microorganisms, the temperature of the pile can be quickly increased and the pH value of the pile can be reduced, shortening the initial time of pile fermentation, but this method takes about 50 days to complete the fermentation, which is a long time period.

[0005] Therefore, the art lacks a bacterial agent formula that can directly solve the problems of slow temperature rise and short high-temperature period in urban manure composting and can achieve rapid fermentation in a short time period. Summary of the invention

[0006] The first aspect of the present invention aims to provide a microbial composition.

[0007] The second aspect of the present invention is to provide a microbial agent.

[0008] The purpose of the third aspect of the present invention is to provide a method for preparing the microbial agent described in the second aspect of the present invention.

[0009] The purpose of the fourth aspect of the present invention is to provide the use of the microbial combination of the first aspect of the present invention and the microbial agent of the second aspect of the present invention.

[0010] In order to achieve the above-mentioned purpose of the present invention, the technical solution adopted by the present invention is:

[0011] The first aspect of the present invention provides a microbial composition, which comprises a mesophilic Bacillus and a thermophilic fungus.

[0012] In some embodiments of the present invention, the mesophilic Bacillus includes but is not limited to at least one of Bacillus Velezii, Bacillus licheniformis, Bacillus amyloliquefaciens, and Bacillus subtilis.

[0013] In some embodiments of the present invention, the thermophilic fungi include Thermomyces lanuginosus, Myceliophthora thermophila, and Thermoascus fragilis; but are not limited to other bacterial agent combinations comprising three thermophilic fungi.

[0014] In some embodiments of the present invention, the microbial composition includes Bacillus Velez, Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus subtilis, Thermomyces lanuginosus, Myceliophthora thermophila, and Thermoascus fragilis.

[0015] The second aspect of the present invention provides a microbial agent comprising the microbial combination described in the first aspect of the present invention.

[0016] In some embodiments of the present invention, the microbial agent further includes an adsorbent and a protective agent.

[0017] In some embodiments of the present invention, the adsorbent includes at least one of vermiculite, wheat bran, oatmeal, rice bran, corn cob powder, and diatomaceous earth.

[0018] In some embodiments of the present invention, the protective agent includes at least one of trehalose, mannitol, glucose, skimmed milk powder, Tween-80, and glycerol.

[0019] The third aspect of the present invention provides a method for preparing the microbial agent according to the second aspect of the present invention, comprising the following steps:

[0020] Preparation of mesophilic Bacillus inoculants;

[0021] Preparation of thermophilic fungal inoculants;

[0022] Mix mesophilic Bacillus inoculants and thermophilic fungal inoculants.

[0023] In some embodiments of the present invention, the preparation of the mesophilic Bacillus agent comprises the following steps:

[0024] 1) Inoculating the seed solution (100 million CFU / mL) of each mesophilic Bacillus into the culture medium in equal volume ratios, the culture conditions are pH 6.8-7.2, stirring intensity 180-200 rpm, aeration volume 1:1-1:1.5, liquid volume <60%, and culturing for 36-48 hours to obtain a fermentation solution;

[0025] 2) Mixing the fermented bacterial liquid of mesophilic Bacillus with the adsorption carrier, shaking at 80-120 rpm for 60-120 minutes, and centrifuging to achieve the immobilization of the strain;

[0026] 3) mixing the immobilized strain with the protective agent, and drying at 50-70° C. until the water content is less than 20%, to obtain a mesophilic Bacillus agent;

[0027] In some embodiments of the present invention, the adsorption carrier includes one or more of vermiculite, wheat bran, oatmeal, rice bran, and corn cob powder;

[0028] In some embodiments of the present invention, the protective agent includes one or more of glucose, skimmed milk powder, Tween-80, mannitol, and glycerol.

[0029] In some embodiments of the present invention, the culture medium may use conventional culture medium in the art (eg, LB culture medium), and may be fed in batches using conventional carbon sources or nitrogen sources in the art such as sucrose, yeast powder, ammonium chloride, sodium nitrate, etc.

[0030] In some embodiments of the present invention, the ratio (m / m) of the fermentation broth to the adsorption carrier is 1:1 to 3.5:1.

[0031] In some embodiments of the present invention, the ratio (m / m) of the protective agent to the immobilized strain is 2.5% to 12.5%.

[0032] In some embodiments of the present invention, the viable count of the mesophilic Bacillus inoculant product is 1*10 10 ~2*10 11 cfu / g;

[0033] In some embodiments of the present invention, the method for preparing the thermophilic fungal agent comprises the following steps:

[0034] 1) preparing a solid-state fermentation substrate, wherein the C / N ratio of the fermentation substrate is 20:1 to 40:1, and the moisture content is 50% to 70%;

[0035] 2) Inoculate the seed solution of thermophilic fungi (100 million CFU / mL) into the solid fermentation substrate and incubate at 40-50°C with an aeration rate of 0.1-0.2 m 3 Air / (min.m 3 Shallow tray mixed fermentation was carried out for 5 to 7 days under the conditions of culture medium;

[0036] 3) After the fermentation is completed, the mixture is dried under ventilation until the moisture content is less than 20% to obtain a thermophilic fungus mixed culture; an adsorbent and a protective agent are added to the thermophilic fungus mixed culture to obtain a thermophilic fungus agent.

[0037] In some embodiments of the present invention, the fermentation substrate comprises one or more of rice bran, wheat bran, corn cob meal, oatmeal, and soybean meal.

[0038] In some embodiments of the present invention, the adsorbent includes at least one of diatomaceous earth and corn cob powder.

[0039] In some embodiments of the present invention, the protective agent includes at least one of trehalose and mannitol.

[0040] In some embodiments of the present invention, the ratio of the mixed culture: adsorbent: protective agent is (20-25):4:1.

[0041] In some embodiments of the present invention, the total number of spores of the thermophilic fungal agent is 2*10 9 ~5*10 10 cfu / g.

[0042] In some embodiments of the present invention, the mass ratio of the mesophilic Bacillus agent to the thermophilic fungus agent is (3:1) to (1:3).

[0043] The fourth aspect of the present invention provides use of the microbial composition described in the first aspect of the present invention and the microbial agent described in the second aspect of the present invention in the preparation of compost fermentation products.

[0044] In some embodiments of the present invention, the uses of the compost fermentation product include but are not limited to urban feces compost fermentation.

[0045] The beneficial effects of the present invention are:

[0046] 1) The present invention is original in its microbial agent formulation.

[0047] Common fermentation agent formulas are mainly based on bacteria, without targeted supplementation of thermophilic fungi, and thermophilic fungi are not used to maintain high temperatures; multiple strains of thermophilic fungi cover multiple functions such as amylase production, cellulase production, and protease production, and can effectively adapt to the types of materials during the high temperature period of fermentation. At the same time, mesophilic Bacillus is helpful for the initial temperature rise, and can resist high temperatures in the form of spores and participate in the cooling and post-ripening stages. In summary, the formula of the agent is a combination of multiple strains of thermophilic fungi with different enzyme-producing functions + several mesophilic Bacillus, and there is no identical literature report on the formula of the agent.

[0048] 2) The preparation method of the present invention avoids the mutual influence between different strains.

[0049] The preparation method of common microbial agents usually first obtains a mixed bacterial solution in proportion, and then prepares a solid microbial agent by carrier adsorption or the like. This method is simpler in terms of process, but mesophilic Bacillus is mostly bacteria, and thermophilic fungi is mostly molds. There are differences in the cultivation methods of the two. In order to increase the concentration of the target strain per unit mass, the present invention adopts the method of "Bacillus and thermophilic fungi" independent preparation-physical mixing, which reduces the mutual influence and ensures the concentration and activity of the two types of microorganisms. Independent preparation can adjust the composition of the agent according to the type of fermentation material, and flexibly adjust the abundance of functional enzymes such as protein enzymes and cellulases. In terms of the separate synthesis of mesophilic Bacillus and thermophilic fungi, a mixed seed solution mixed fermentation method is adopted, and the above two agents are prepared in one step. There is innovation in the preparation process, and not all strains are prepared separately and then compounded. A specific ratio is formed inside the two agents of mesophilic Bacillus and thermophilic fungi, which can obtain a higher growth rate.

[0050] 3) The present invention has achieved better technical effects.

[0051] The bacterial agent of the present invention can quickly heat up, the high temperature period is longer, and a round of fermentation can be completed in a shorter time period. The present invention shows through experimental data that the bacterial agent prepared by the present invention can reach a high temperature of 75°C within 2 days and maintain a high temperature state until the 10th day. The fermentation cycle is short, and a round of fermentation only takes 16 days. The high temperature period can occupy about 56% to 62.5% of the entire fermentation cycle. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0053] Figure 1 The figures are the fermentation temperature variation results of different bacterial fermentation combinations of the present invention.

[0054] Figure 2 The figure is a comparison of the fermentation temperature changes of the bacterial agent of the present invention and the commercial bacterial agent. DETAILED DESCRIPTION

[0055] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0056] The strain source information adopted by the present invention is as follows:

[0057] Thermophilic fungi: It consists of multiple strains of thermophilic fungi, including Thermomyces lanuginosus (CICC 41717), Myceliophthora thermophila (CICC 2441), and Thermoascuscrustaceus (CICC 41719).

[0058] Mesophilic Bacillus: Bacillus velezensis (CICC 24528), Bacillus licheniformis (CICC 24403), Bacillus megaterium (CICC 24803), Bacillus amyloliquefaciens (CICC 10036), Bacillus subtilis (CICC 24341).

[0059] The above strains can all be purchased from the China Industrial Microbiological Culture Collection Center (CICC).

[0060] Example 1 Thermophilic fungus agent

[0061] In some embodiments of the present invention, a method for preparing a thermophilic fungal agent with better effect comprises the following steps:

[0062] 1) Activated strains: Three strains (Thermomyces lanuginosus (CICC41717), Myceliophthora thermophila (CICC 2441), and Thermoascus crustaceus (CICC 41719)) were purchased from the China Industrial Microbiological Culture Collection Center; a slant stored at 4°C was taken, and the strains were placed in the center area of ​​a PDA solid culture medium and cultured upright in an incubator at 45°C.

[0063] 2) Preparation of seed solution: Pick colonies on PDA solid culture medium, inoculate into sterilized PDA liquid culture medium (liquid volume 100 mL / 250 mL), and culture in a shaking flask in a 45° C. shaking incubator for 5 days to obtain seed solution.

[0064] 3) Preparation of solid culture medium: using wheat bran, rice bran, corn cob powder and soybean meal in a mass ratio of 3:2:1:1 as the base, C / N of 25, and using sterile deionized water to adjust the above mixed materials to an initial moisture content of 60% to prepare a solid fermentation medium.

[0065] 4) Shallow plate expansion culture: Inoculate the seed suspension (100 million CFU / mL) of two strains of Thermoascus brittletus and Thermoascus lanuginosus into the solid fermentation medium in equal volume ratio, with the seed liquid inoculation ratio of 5‰ to 1%, stir evenly and place in the fermentation box, at 45°C and aeration rate of 0.1 to 0.2 m 3 Air / (min.m 3 The culture medium was mixed and fermented in a shallow dish for 4 days, with sterile deionized water added regularly to maintain the moisture content of the culture medium. Then, the seed solution of Thermomycetes pyrifolia (100 million CFU / mL) was inoculated into the solid fermentation medium, and the culture conditions remained unchanged for 5 to 7 days. The initial inoculation ratio of the seeds of the three strains was the same.

[0066] 5) After fermentation, ventilate and dry until the moisture content is less than 20%, crush, sieve (80 mesh), and package to obtain a mixed culture of thermophilic fungi; add an adsorbent (diatomaceous earth) and a protective agent (trehalose: mannitol = 10:1) to the pure culture of the mixed strains, and keep the mass ratio of bacterial powder culture: adsorbent: protective agent at 25:4:1 to obtain a mixed culture of thermophilic fungi.

[0067] Among them, the total number of spores of thermophilic fungal agents is 2*10 9 ~5*10 10 cfu / g.

[0068] Example 2: Cotton-like thermophilic fungus agent

[0069] (1) As in Example 1, the strain of Thermomyces lanuginosus was activated to prepare a seed suspension, and the Thermomyces lanuginosus was inoculated alone;

[0070] (2) As in Example 1, a solid-state fermentation medium was prepared using wheat bran, rice bran, corn cob powder, and soybean meal in a mass ratio of 3:2:1:1, a C / N ratio of 25, and sterile deionized water to adjust the above-mentioned mixture to an initial moisture content of 60%;

[0071] (3) The inoculation ratio of the sparse cotton-like thermophilic mold strain is 5‰ to 1%, and it is evenly stirred and placed in the fermentation box at 45°C and a ventilation rate of 0.1 to 0.2 m 3 Air / (min.m 3 Medium), regularly supplemented with sterile deionized water to maintain the water content of the medium, and then mixed fermentation was carried out in a shallow tray for 7 days;

[0072] (4) After fermentation, the mixture is ventilated and dried until the moisture content is less than 20%, and then crushed, sieved (80 mesh), and packaged to obtain a sparse cotton-like thermophilic mold culture; an adsorbent (diatomaceous earth) and a protective agent (trehalose: mannitol = 10:1) are added to the mixed strain pure culture, and the mass ratio of the bacterial powder culture: adsorbent: protective agent is maintained at 25:4:1, thereby obtaining a sparse cotton-like thermophilic mold agent.

[0073] Among them, the total number of spores of the sparse cotton-like thermophilic mold agent is 2*10 9 ~5*10 10 cfu / g.

[0074] Example 3 Thermomycetes fungus agent

[0075] (1) As in Example 1, Myceliophthora thermophila was activated, a seed suspension was prepared, and Myceliophthora thermophila was inoculated alone.

[0076] (2) As in Example 1, a solid-state fermentation medium was prepared using wheat bran, rice bran, corn cob powder, and soybean meal in a mass ratio of 3:2:1:1, a C / N ratio of 25, and sterile deionized water to adjust the above-mentioned mixture to an initial moisture content of 60%.

[0077] (3) The inoculation ratio of the thermophilic Myceliophthora strain is 5‰ to 1%, which is evenly stirred and placed in a fermentation box at 45°C and a ventilation rate of 0.1 to 0.2 m 3 Air / (min.m 3 The culture medium was mixed and fermented in a shallow tray for 7 days under the conditions of regularly adding sterile deionized water to maintain the moisture content of the culture medium.

[0078] (4) After fermentation, ventilate and dry until the moisture content is less than 20%, crush, sieve (80 mesh), and package to obtain the thermophilic myceliophthora culture; add adsorbent (diatomaceous earth) and protective agent (trehalose: mannitol = 10:1) to the mixed strain pure culture, and keep the mass ratio of bacterial powder culture: adsorbent: protective agent at 25:4:1 to obtain the thermophilic myceliophthora inoculum. The total number of spores of the thermophilic myceliophthora inoculum is 2*10 9 ~5*10 10 cfu / g.

[0079] Example 4: Thermoascus brittle thermoscomycetes agent

[0080] (1) As in Example 1, Thermoascus fragilis was activated, a seed suspension was prepared, and Thermoascus fragilis was inoculated alone.

[0081] (2) As in Example 1, a solid-state fermentation medium was prepared using wheat bran, rice bran, corn cob powder, and soybean meal in a mass ratio of 3:2:1:1, a C / N ratio of 25, and sterile deionized water to adjust the above-mentioned mixture to an initial moisture content of 60%.

[0082] (3) The inoculation ratio of the brittle thermophilic ascomycete strain is 5‰ to 1%, stirred evenly and placed in a fermentation box at 45°C and a ventilation rate of 0.1 to 0.2 m 3 Air / (min.m 3 The culture medium was mixed and fermented in a shallow tray for 7 days under the conditions of regularly adding sterile deionized water to maintain the moisture content of the culture medium.

[0083] (4) After fermentation, ventilate and dry until the moisture content is less than 20%, crush, sieve (80 mesh), and package to obtain the brittle thermophilic ascomycete culture; add an adsorbent (diatomaceous earth) and a protective agent (trehalose: mannitol = 10:1) to the mixed strain pure culture, and maintain the mass ratio of bacterial powder culture: adsorbent: protective agent at 25:4:1 to obtain the brittle thermophilic ascomycete culture agent.

[0084] Among them, the total number of spores of Thermoascus brittleus is 2*10 9 ~5*10 10 cfu / g.

[0085] Example 5 Mesophilic Bacillus Agent

[0086] (1) Activated strains: Each strain (Bacillus Velez, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, and Bacillus subtilis) purchased from the China Industrial Microbiological Culture Collection was streaked onto an LB plate and cultured at 35°C for 24 h.

[0087] (2) Preparation of seed solution: Pick a colony from the LB plate, inoculate it into sterilized LB liquid medium (liquid volume 100 mL / 250 mL), place it in a shaking incubator at 35°C, 190 rpm, and culture for 24 h to obtain seed solution with a seed solution concentration of 100 million CFU / mL;

[0088] (3) Fermentation tank expansion culture:

[0089] 1) The seed liquid of each mesophilic Bacillus strain was inoculated into LB medium in equal volume ratio, and the volume of the seed liquid was 1% to 2% of the volume of the LB medium;

[0090] 2) The culture conditions were pH 7.0, stirring intensity 190 rpm, aeration volume 1:1.25, and liquid volume <60%;

[0091] 3) Determine the concentration of carbon source and nitrogen source during the fermentation process, calculated as COD and total nitrogen, respectively. When the concentration exceeds 50% of the initial concentration, perform batch feeding operation with sucrose or sodium nitrate as the carbon source or nitrogen source, and restore to the original concentration to continue fermentation;

[0092] 4) Detect the absorbance value (OD) of LB culture medium 600 ) until stable, and then the fermentation is terminated to obtain a mesophilic Bacillus mixed bacterial liquid;

[0093] (4) Preparation of mesophilic Bacillus solid bacterial agent

[0094] 1) Wheat bran and corncob powder in a mass ratio of 1:1 were used as composite adsorption carriers, added to the mixed fermentation bacterial solution for low-speed oscillation adsorption (100 rpm, 90 min) and high-speed centrifugation (4000-6000 rpm, 30 min), and the mass ratio of the adsorption carrier to the bacterial solution was 1:3 to achieve the immobilization of the mixed strains;

[0095] 2) Add a protective agent (glucose: skimmed milk powder: Tween-80 = 2:10:1) at a mass ratio of 3% to the immobilized bacterial agent, mix well, and dry at 60°C for 10 hours. The moisture content is <20%, and a mesophilic Bacillus mixed bacterial agent can be obtained.

[0096] The number of live bacteria in the mesophilic Bacillus inoculant product is 1*10 10 ~2*10 11 cfu / g.

[0097] Example 6 Mixed bacterial agent

[0098] The thermophilic spore fungus agent obtained in Example 1 and the mesophilic bacillus mixed agent obtained in Example 5 were mixed in a mass ratio of 1:1 to obtain.

[0099] Example 7 Mixed bacterial agent

[0100] The sparse cotton-like thermophilic mold agent obtained in Example 2 and the mesophilic Bacillus mixed agent obtained in Example 5 were mixed in a mass ratio of 1:1 to obtain.

[0101] Example 8 Mixed bacterial agent

[0102] The thermophilic Myceliophthora inoculum obtained in Example 3 and the mesophilic Bacillus mixed inoculum obtained in Example 5 were mixed in a mass ratio of 1:1 to obtain.

[0103] Example 9 Mixed bacterial agent

[0104] The Thermoascus fragilis bacterial agent obtained in Example 4 and the Mesophilic Bacillus mixed bacterial agent obtained in Example 5 were mixed in a mass ratio of 1:1 to obtain.

[0105] Effect Example 1

[0106] The composting box (40cm*30cm*20cm) was used as a reactor. The composting raw materials were mainly urban feces after flocculation and dehydration, and vegetarian residue and sawdust (1:1) were used as auxiliary materials. The ratio of the main material to the auxiliary material was 4:6, and the initial moisture content was 65%. The fermentation temperature verification experiment was carried out according to different microbial agent formulas.

[0107] The specific combinations are as follows:

[0108] A1: mesophilic Bacillus agent + thermophilic spore fungus agent (mass ratio 1:1, prepared in Example 6).

[0109] A2: mesophilic Bacillus agent + sparse cotton-like thermophilic mold agent (mass ratio 1:1, prepared in Example 7).

[0110] A3: mesophilic Bacillus agent + thermophilic Myceliophthora agent (mass ratio 1:1, prepared in Example 8).

[0111] A4: mesophilic Bacillus agent + brittle thermoascomycete agent (mass ratio 1:1, prepared in Example 9).

[0112] B1: Mesophilic Bacillus agent.

[0113] B2: Thermophilic spore-forming fungi agent.

[0114] Add microbial agents according to 1‰ of the total mass of the fermented material and mix and stir evenly. The mixture is placed in a composting box for composting experiments. An aeration plate is arranged at the bottom of the composting box to intermittently aerate the pile (aeration for 10 minutes every 30 minutes, and the aeration volume is 4L / min). When the temperature of the mixed fermentation pile drops to less than 40-45°C, it is considered that the fermentation is completed. The fermentation temperature rise data of each group are shown in Figure 2. Figure 1 shown.

[0115] from Figure 1 The results show that the A1 combination can reach a high temperature of 70°C on the second day and can maintain it until the 10th day, and it only takes 16 days to complete a fermentation cycle. The A2, A3, and A4 combinations can also complete fermentation in 22 to 26 days, with the highest temperature close to 70°C, but the effects of the B1 and B2 combinations are poor. The highest temperature of B1 can only reach 65°C, and as of the end of the statistical time, it has not completed a fermentation cycle. Although the highest temperature of B2 can reach 70°C, it took 18 days to reach the temperature peak, and as of the end of the statistical time, it has not completed a fermentation cycle.

[0116] It can be seen that the combination of A1 to A4 can make the compost fermentation quickly reach the temperature peak and shorten the entire fermentation process.

[0117] Effect Example 2

[0118] The composting box (40cm*30cm*20cm) is used as a reactor. The composting raw materials are mainly urban feces after flocculation and dehydration, and bamboo residue and sawdust (1:1) are auxiliary materials. The ratio of the main material to the auxiliary material is 4:6, and the initial moisture content is 65%. The effects of the A1 combination of the present invention and the commercial microbial agent are compared. The commercial microbial agent is purchased from a company in Hebi City. The microbial agent composition includes a variety of beneficial microorganisms such as highly active Bacillus, yeast, and Enterococcus faecalis.

[0119] Add microbial agents according to 0.5‰ of the total mass of the fermented material and mix and stir evenly. The mixture is placed in a composting box for composting experiments. An aeration plate is arranged at the bottom of the composting box to intermittently aerate the pile (aeration for 10 minutes every 30 minutes, and the aeration volume is 4L / min). When the temperature of the mixed fermentation pile drops to less than 40-45°C, it is considered that the fermentation is completed. The fermentation temperature rise data of group A1 and the commercial microbial agent group are shown in Figure 2 shown.

[0120] It can be seen that the A1 combination can heat up faster than the commercial bacterial agent. The fermentation temperature was detected to be above 55°C on the second day, and the highest temperature was above 70°C on the third day. The temperature of the homemade composite bacterial agent was higher than 60°C from day 2 to 11, with a longer high temperature period. The homemade bacterial agent detected that the pile temperature was between 40 and 45°C on the 17th day, which means that the first fermentation stage was completed. The commercial bacterial agent heated up to above 60°C on the 6th day, reached the highest fermentation temperature of 65°C on the 11th day, and the fermentation temperature was higher than 60°C from day 6 to 17. The commercial bacterial agent detected that the pile temperature was between 40 and 45°C on the 29th day of the experiment, completing a fermentation.

[0121] In summary, the present invention adopts a unique bacterial agent formula, which effectively solves the problems of slow temperature rise and short high temperature period in high temperature fermentation. It adopts a mode of mixed fermentation of multiple mesophilic Bacillus and mixed fermentation of multiple thermophilic fungi, and independently prepares and then mixes them, which ensures the activity of different types of strains. The growth rate is faster in actual use, and rapid temperature rise can be performed in experiments.

Claims

1. A microbial composition, characterized in that: The microbial composition comprises mesophilic bacillus and thermophilic fungi.

2. The microbial composition according to claim 1, characterized in that: The mesophilic Bacillus includes but is not limited to at least one of Bacillus Velezii, Bacillus licheniformis, Bacillus amyloliquefaciens and Bacillus subtilis.

3. The microbial composition according to claim 1, characterized in that: The thermophilic fungi include Thermomyces lanuginosus, Thermomyces thermophilus, and Thermoascus fragilis; Preferably, other thermophilic fungi may also be included.

4. A microbial agent, characterized in that: The bacterial agent comprises the microbial combination according to claim 1.

5. The microbial agent according to claim 4, characterized in that: The microbial agent also includes an adsorbent and a protective agent.

6. The method for preparing the microbial agent according to claim 4 or 5, comprising the following steps: Preparation of mesophilic Bacillus inoculants; Preparation of thermophilic fungal inoculants; Mix mesophilic Bacillus inoculants and thermophilic fungal inoculants.

7. The preparation method according to claim 6, characterized in that: The preparation of the mesophilic bacillus agent comprises the following steps: 1) Inoculating the seed liquid of mesophilic Bacillus into the culture medium, the culture conditions are 35° C., pH 6.8-7.2, stirring intensity 180-200 rpm, aeration volume 1:1-1:1.5, liquid volume <60%, culturing for 36-48 hours, and obtaining a fermentation liquid; 2) Mixing the fermented bacterial liquid of mesophilic Bacillus with the adsorption carrier, shaking at 80-120 rpm for 60-120 minutes, and centrifuging to achieve the immobilization of the strain; 3) mixing the immobilized strain with the protective agent, drying at 50-70° C. until the water content is less than 20%, to obtain a mesophilic Bacillus agent; Preferably, the adsorption carrier comprises one or more of vermiculite, wheat bran, oatmeal, rice bran, and corn cob powder; Preferably, the protective agent includes one or more of glucose, skimmed milk powder, Tween-80, mannitol, and glycerol.

8. The preparation method according to claim 6, characterized in that: The preparation of the thermophilic fungal agent comprises the following steps: 1) preparing a solid-state fermentation substrate, wherein the C / N ratio of the fermentation substrate is 20:1 to 40:1 and the moisture content is 50% to 70%; 2) Inoculate the seed liquid of thermophilic fungi into the solid fermentation substrate, at 40-50°C and aeration rate of 0.1-0.2 m 3 Air / (min.m 3 Shallow tray mixed fermentation was carried out for 5 to 7 days under the conditions of culture medium; 3) After fermentation, ventilate and dry until the moisture content is less than 20% to obtain a thermophilic fungus mixed culture; add an adsorbent and a protective agent to the thermophilic fungus mixed culture to obtain a thermophilic fungus agent; Preferably, the fermentation substrate comprises one or more of rice bran, wheat bran, corn cob meal, oatmeal, and soybean meal; Preferably, the adsorbent comprises at least one of diatomaceous earth and corn cob powder; Preferably, the protective agent includes at least one of trehalose and mannitol.

9. The preparation method according to claim 6, characterized in that: The mass ratio of the mesophilic bacillus agent to the thermophilic fungus agent is (3:1) to (1:3).

10. Use of the microbial composition according to any one of claims 1 to 3 or the microbial agent according to any one of claims 4 or 5 in the preparation of compost fermentation products.

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