Compost decomposition agent and application thereof

Abstract

The invention relates to the technical field of functional microorganism screening and application, in particular to a composting agent capable of composting at low temperature, normal temperature and medium-high temperature as well as a preparation method and application thereof. The decomposition agent comprises an active component and an adsorption carrier; wherein the active ingredients are lysine bacillus, klebsiella and enterobacter; the rice hull biochar is used as a carrier, and the obtained compound microbial agent is a powder microbial agent and can be used for composting and aeration in different temperature environments such as low temperature, normal temperature and medium-high temperature. By screening and optimizing strains and combining with reasonable formula design, the activity of the decomposition agent in low-temperature, normal-temperature and medium-high-temperature environments is remarkably improved, the technical problems of narrow applicable temperature range, low efficiency, poor product quality, potential environmental risk and the like of the decomposition agent in the prior art are solved, annual decomposition can be realized, and the decomposition efficiency is improved. The method is not affected by seasons, resources are fully utilized, and the resource utilization efficiency is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of functional microorganism screening and application, and in particular to a composting agent capable of composting at low temperature, normal temperature or medium or high temperature, and a preparation method and application thereof. Background Art

[0002] Agricultural solid waste refers to solid waste generated during agricultural production and processing, mainly including crop straw, livestock and poultry manure, agricultural film, pesticide packaging, etc. These wastes are rich in organic matter, such as sugars, proteins, amino acids, inorganic salts, starch, cellulose, etc., and are important renewable resources. However, my country's agricultural solid waste output is huge, with an annual output of more than 5 billion tons, of which the comprehensive utilization rate of livestock and poultry manure is less than 60%. The accumulation of a large amount of waste not only causes a waste of resources, but also causes serious pollution to the ecological environment.

[0003] Organic waste composting is an important method for treating organic waste. It converts organic waste into organic fertilizer by using microorganisms such as bacteria, fungi, and actinomycetes to achieve resource recycling. However, there are some problems with traditional composting methods, such as long composting cycle, incomplete decomposition, and poor product quality. In recent years, with the increasing emphasis on environmental protection, people have an increasing demand for efficient and environmentally friendly composting technology.

[0004] In the prior art, some composting agents can improve composting efficiency within a specific temperature range, but the applicable temperature range is limited. For example, psychrophilic composting agents can only work in low temperature environments, and thermophilic composting agents can only work in high temperature environments. In addition, some composting agents contain chemical additives that can pollute the environment. Therefore, it is particularly important to develop a biological composting agent that can effectively promote composting under different temperature conditions. Summary of the invention

[0005] In order to overcome the technical problems of the prior art such as narrow applicable temperature range, low efficiency, poor product quality and potential environmental risks of the decomposing agent, the present invention provides a decomposing agent capable of composting at low temperature, room temperature and medium to high temperature, and a preparation method and application thereof.

[0006] To achieve the above-mentioned purpose of the present invention, the following technical solutions are used:

[0007] A composting agent, the composting agent is an active ingredient and an adsorption carrier; wherein the active ingredients are lysine bacillus, Klebsiella, and Enterobacter; wherein,

[0008] Lysinibacillus sp. LFS1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32336;

[0009] Enterobacter sp. ENT1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32337;

[0010] Klebsiella VHK1 (Klebsiella sp.) was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, and the deposit number is CGMCC No: 32339.

[0011] The volume mass ratio of the active ingredient to the adsorption carrier is (1-5):1.

[0012] The volume ratio of the bacterial suspensions of Bacillus lysinicola, Klebsiella and Enterobacter in the active ingredient is (1-5): (1-3): (1-2).

[0013] The active ingredients are Bacillus lysine, Klebsiella, and Enterobacter, which are placed in LB medium and cultured to the logarithmic growth phase, and the precipitates are collected by centrifugation. Each precipitate is resuspended in sterile saline to OD 600 The bacterial suspension is 0.8-1.2, and then the bacterial suspensions are mixed in a volume ratio of (1-5): (1-3): (1-2), and then fermented in LB medium to obtain a composite fermentation liquid.

[0014] The bacterial suspensions of lysine bacillus, Klebsiella and Enterobacter in the active ingredients are in a volume ratio of (1-3):(1-2):(1-2).

[0015] The adsorption carrier is rice husk biochar.

[0016] An application of the composting agent, and an application of the composting agent in preparing organic fertilizer by composting.

[0017] The composting agent is used in preparing organic fertilizer by composting under different initial conditions of low temperature, normal temperature or medium and high temperature.

[0018] The composting agent is used in starting composting to prepare organic fertilizer under different initial environmental conditions of low temperature (5°C-20°C), normal temperature (20°C-30°C) or medium-high temperature (above 30°C).

[0019] The composting agent is added to agricultural solid waste for composting at different initial temperatures, such as low temperature (5°C-20°C), normal temperature (20°C-30°C) or medium-high temperature (above 30°C).

[0020] The agricultural solid waste is any one of cow dung and straw, or a mixture of the two.

[0021] The advantages of the present invention are:

[0022] The composting agent of the present invention is composed of spindle-shaped lysine bacillus, Klebsiella mutans and Enterobacter, and is suitable for carrying out normal composting fermentation work of livestock and poultry manure in different temperature environments (10-30°C). A biological composting agent that can effectively promote composting under different temperature conditions is developed to solve the limitations of the existing technology, improve composting efficiency, shorten the composting cycle, and reduce composting costs.

[0023] Cow dung has low nutrient content and high cellulose and lignin content. The main microbial strain of the decomposition agent of the present invention is resistant to low temperature, room temperature and medium and high temperature and has the ability to degrade cellulose and lignin. The strain not only has a wide temperature adaptability range, but also has strong ability to degrade cellulose and lignin at different temperatures of 10°C, 20°C and 30°C, and can be used to accelerate the decomposition speed of organic matter and promote composting.

[0024] The performance of strains at different temperatures is different. Some strains have specific degradation functions such as cellulose degradation only at room temperature, while other strains have corresponding functions only at high temperatures. There are relatively few strains that can tolerate both low temperatures and medium and high temperatures, and there are even fewer strains that are resistant to low temperatures, room temperatures, and medium and high temperatures and have good and strong cellulose degradation capabilities at different temperatures. The main microorganisms in the composting agent of the present invention have high cellulose and lignin degradation capabilities at 10°C, 20°C, and 30°C at the same time.

[0025] The main microorganisms of the composting agent of the present invention grow on LB basic medium, and the colony morphology is shown in Figure 1 .

[0026] Through 16S rDNA molecular identification of the decomposition bacteria, the strains were determined to be fusiform lysinibacillus (FLB), variable Klebsiella (VHK) and Enterobacter (EG).

[0027] In another aspect, the decomposition agent of the present invention is combined with rice husk biochar to prepare a decomposition agent biological agent, which is used or transported in the form of a solid agent. In view of the fact that the strain of the present invention has the ability to degrade cellulose and lignin under different temperature conditions, it can quickly decompose macromolecular organic matter, decompose organic materials, and can be used as a decomposition agent for organic materials. In particular, because the decomposition agent has a good and strong degradation ability at different temperatures. Therefore, it can effectively promote the decomposition of compost under different temperature conditions. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0029] Figure 1 Antagonistic effect diagram between strains provided by the present invention

[0030] Figure 2 This is the colony morphology of the main components of the decomposition bacteria agent of the present invention

[0031] Figure 3 Results of the ability of some strains provided by the present invention to degrade cellulose on CMC culture medium at different temperatures

[0032] Figure 4 Results of the ability of some strains provided by the present invention to degrade lignin on aniline blue medium at different temperatures

[0033] Figure 5 The results of the cellulose degradation ability of different strains in the comparative example provided by the present invention in combination with each other on CMC culture medium

[0034] Figure 6 The results of the ability of different strains in pairs to degrade cellulose on aniline blue medium in the comparative example provided by the present invention

[0035] Figure 7 The results of the cellulose degradation ability of three-three combinations of different strains on CMC culture medium in the comparative example provided by the present invention

[0036] Figure 8 The results of the ability of different strains in three combinations to degrade cellulose on aniline blue medium in the comparative example provided by the present invention

[0037] Fig. 9 Temperature changes of compost during composting

[0038] Fig.10 Effect of composting agent on the composting of cow dung mixture at different temperatures

[0039] Fig.11 Changes of average root length of carrot seeds and germination index of compost seeds under different temperatures DETAILED DESCRIPTION

[0040] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0041] The composting agent of the present invention has strong adaptability and can be widely used in different climate regions and seasons; it can operate efficiently around the clock, and can maintain efficient decomposition effect even in low temperature environment, without additional heating, saving energy consumption; it can flexibly respond to temperature changes, adapt to temperature fluctuations, and can stably play a role even in an environment with a large temperature difference between day and night. The decomposition agent of the present invention has high decomposition efficiency, which can accelerate the decomposition of organic matter, promote the synergistic effect of microbial flora, quickly decompose organic waste, shorten the composting cycle, and improve resource utilization; significantly reduce composting time, reduce composting cycle, reduce manpower and material costs, and improve resource recycling efficiency. And the decomposition product of the decomposition agent of the present invention is of good quality, and the decomposition product contains rich humus, which can effectively improve soil structure, enhance soil fertility, and improve soil water and fertilizer retention capacity; organic fertilizer is rich in various nutrients required for plant growth, can promote plant growth, and improve crop yield and quality; microbial flora can effectively degrade harmful substances such as pathogens, insect eggs, and heavy metals in organic waste, and improve the safety of organic fertilizer.

[0042] If the specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or conditions recommended by the manufacturer. If the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0043] Example 1

[0044] 1) Isolation and identification of strains:

[0045] Take a low-temperature cow dung sample stored in a refrigerator at 0-4°C for 6 months, and use the dilution separation method to separate microorganisms in a cow dung juice solid culture medium (cow dung juice 500mL, glucose 10g, yeast extract 2.5g, sodium chloride 5g, agar 15g, distilled water to freely adjust the pH value to 7.0±0.2, the final volume is adjusted to 1L, and sterilized at 121°C for 30min). Culture it at 0-4°C, pick the strain of the fastest growing microbial colony within 48h, and then use broth culture medium (ingredients: beef extract 5.0g, peptone 10.0g, NaCl 10.0g, agar 20.0g, distilled water to 1000mL, pH: 7.0, sterilized at 121°C for 30min) to streak and purify it under 0-4°C culture conditions, inoculate it into a 10mL broth test tube, and culture it at 10°C. OD was measured at 6h, 12h, and 24h of culture. 600The growth rates of the individual strains were compared by comparing the values, and the fastest growing strain was retained. The strain was then purified and inoculated into a sterilized broth solid medium and cultured at 0-4°C for 24 hours. The strain was then acclimated at different temperatures of 10°C, 15°C, 20°C, 25°C, 30°C, and 35°C.

[0046] 2) Screening of microbial strains that degrade cellulose

[0047] Take 10g of a mid-fermentation sample of a cow dung pile near a cow farm in Liaoning, add 90mL of sterile water, shake thoroughly, and let stand for 2h. Take 1mL of the supernatant and dilute it to 10mL with sterile water to make a 10-1 dilution. Use a pipette to mix well, then take 1mL and dilute it to 10mL with sterile water to make a 10-1 dilution. -2 Dilute to 10 times the dilution as above -9 times. Take 10 -1 , 10 -3 , 10 -5 , 10 -7 , 10 -9 100 μL of the five dilutions were cultured and observed in broth solid medium (5.0 g beef extract, 10.0 g peptone, 10.0 g NaCl, 20.0 g agar, distilled water to 1000 mL, pH: 7.0, autoclaved at 121°C for 30 min) and Martin's solid medium (200 g fresh potato peeled, cut into small pieces, boiled for 30 min, filtered, 20 g glucose added, distilled water to 1 L, 20.0 g agar added, heated and melted, autoclaved at 121°C for 30 min). Single colonies were picked according to the morphological characteristics of the colonies and repeatedly streaked on the solid medium, so that the strains in the culture medium could be separated and purified, and finally a single strain was obtained; the selected single strain was inoculated into sodium carboxymethyl cellulose (CMC-Na) solid medium (1.0 g potassium hydrogen phosphate, 0.25 g magnesium sulfate heptahydrate, 2.0 g yeast powder, 10.0 g agar powder, sodium carboxymethyl cellulose CMC 2.0g, 1000mL water) were placed at 5℃, 15℃, and 25℃ for 2 days, and then stained and washed with sterile methyl red. The transparent zone produced by the strain that can degrade CMC at 5℃ was measured and the results were recorded. The larger the transparent zone produced by the strain, the better the ability to degrade cellulose.

[0048] 3) Antagonistic function test between strains

[0049] The five strains (LFS1, ENT1, VHK1, BTS1, LVB1) screened above were cultured on a broth solid substrate for three days, and the growth of the crossover strains was observed to see if there was an inhibition zone. The larger the inhibition zone, the stronger the antagonism between the strains. After the antagonistic function test, it was found that an inhibition zone was generated between VHK1 and BTS1, and an inhibition zone was also generated between LVB1 and BTS1 strains, and the inhibition zone was larger. LFS1, ENT1, and VHK1 do not produce antagonistic functions to each other. Figure 1 This is a diagram showing the antagonistic function test results among five strains.

[0050] 4) Identification of functional strains

[0051] After the above screening, three strains with strong degradation and no antagonism to each other (LFS1, ENT1, VHK1) were selected for functional strain identification. 16S rDNA was extracted and PCR amplified by colony PCR method. The amplified product was recovered and purified and sent to Shanghai Shenggong Biotechnology Co., Ltd. for sequencing. The sequencing results were BLASTed with the known sequences in the GenBank database to determine the types of strains: LFS1, ENT1, VHK1. PCR amplification of 16rRNA gene was performed, and the LFS1 sequence is shown below:

[0052]

[0053] ENT1 is:

[0054]

[0055] VHK1 is:

[0056]

[0057] The above strains were deposited separately:

[0058] Lysinibacillus sp. LFS1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32336;

[0059] Enterobacter sp. ENT1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32337;

[0060] Klebsiella VHK1 (Klebsiella sp.) was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, and the deposit number is CGMCC No: 32339.

[0061] Example 2: Preparation of composite composting agent

[0062] 1. Preparation of composite fermentation broth: The strains obtained by the above screening were cultured in broth medium (5.0 g beef extract, 10.0 g peptone, 10.0 g NaCl, distilled water to 1000 mL, pH: 7.0) at 10°C, 20°C, and 30°C respectively until the logarithmic growth phase of each strain; then the strains grown to the logarithmic growth phase were centrifuged at 3500 rpm for 10 min to collect the precipitates, and the precipitates were resuspended in sterile saline to OD 600 The single bacterial inoculum was obtained from a bacterial suspension of 1.2 (where LFS1 OD 600 =0.998, ENT1 OD 600 =1.087, VHK1 OD 600 =0.982);

[0063] Then, the bacterial suspensions of LFS1, ENT1, and VHK1 were mixed at a volume ratio of 3:1:2; 4:1:1; 5:3:2; 1:3:2; 1:2:1; 2:1:1; (see Table 1 for the corresponding numbers of each ratio), and then cultured in broth medium (5.0 g beef extract, 10.0 g peptone, 10.0 g NaCl, distilled water to 1 L, pH: 7.0, autoclaved at 121°C for 30 min) at different temperatures of 10°C, 20°C, and 30°C until the logarithmic growth phase, and the composite fermentation broth was obtained. The precipitates were collected by centrifugation at 3500 rpm for 10 min, and the precipitates were resuspended in sterile saline to OD 600The bacterial suspension was 1.0, and was inoculated on sodium carboxymethyl cellulose (CMC-Na) solid culture medium (1.0g potassium hydrogen phosphate, 0.25g magnesium sulfate heptahydrate, 2.0g yeast powder, 10.0g agar powder, 2.0g sodium carboxymethyl cellulose CMC, 1000mL water) and cultured at 10℃, 20℃, and 30℃ for 2d, and then stained and washed with sterile methyl red. The transparent circles produced by the degradation at different temperatures were measured and the results were recorded. The larger the transparent circle produced by the strain, the better the ability to degrade cellulose. Among them, LFS1:ENT1:VHK1=3:1:2 has the best effect, 1:3:2 has the second best effect, and 1:2:1 has the best effect. The ability of the composite bacterial agent to degrade lignin was tested by spotting it on aniline blue decolorization medium (10 g yeast extract, 10 g glucose, 0.1 g aniline blue, 20 g agar, add distilled water to 1000 mL, and sterilize at 115°C for 30 min), culturing at 10°C, 20°C, and 30°C for 48 h, and selecting the one with obvious decolorization circle, which had strong lignin degradation ability, and recording it.

[0064] Table 1 Treatment number of each proportion of bacterial agent

[0065] Bacterial agent number LFS1 ENT1 VHK1 With or without biochar loading 1 3 1 2 none 2 4 1 1 none 3 5 3 2 none 4 1 3 2 none 5 1 2 1 none 6 2 1 1 none 7 3 1 2 have 8 4 1 1 have 9 5 3 2 have 10 1 3 2 have 11 1 2 1 have 12 2 1 1 have

[0066] 2. Carrier adsorption: After the above composite fermentation liquid is tested for qualified viable bacteria, it is mixed with rice husk biochar powder that has passed a 40-mesh sieve at a volume-to-mass ratio (L / kg) of 3:1 for full adsorption (see Table 1 for the number of biochar loaded) and adjusted to a moisture content of ≤25% to obtain a compost rapid decomposition agent. The biochar loaded with the strain was inoculated on CMC-Na medium and aniline blue medium to observe the ability to degrade cellulose and lignin. Figure 3 This is a diagram showing the ability of composite bacterial agents with different proportions and biochar-loaded decomposition agents to degrade cellulose at different temperatures. Figure 4 The following is a graph showing the ability of the decomposing agent to degrade lignin at different temperatures after different proportions of composite bacteria and biochar loading. 3. The prepared decomposing agent is dried, crushed, and packaged into bags for use.

[0067] By the above Figure 3 and Figure 4 It can be seen that at 30°C, the cellulose capacity of LFS1, ENT1, and VHK1 with a volume ratio of 3:1:2 is stronger than that at 20°C and 10°C, and the degradation effect is more prominent after loading biochar. Figure 4 The medium biochar-loaded bacterial liquid treatment (LFS1, ENT1, VHK1 volume ratio of 3:1:2) had the best ability to degrade lignin.

[0068] Comparative Example 1

[0069] The strains obtained by the above screening were mixed with Bacillus amyloliquefaciens (commercially available with high cellulase production activity, named strain A) to carry out cellulose degradation and lignin degradation experiments, specifically:

[0070] The single bacterial inoculant was the above four single strains LFS1, ENT1, VHK1, and strain A, which were inoculated into sterile beef extract peptone medium, placed in a constant temperature shaker at 25°C and 180 r / min for 48 h until the logarithmic phase, and then washed with sterile saline to obtain an OD 600 The bacterial suspension with an OD of 1.0 was the single bacterial agent. The LFS1, ENT1, and VHK1 obtained above were mixed with strain A in equal proportions (1:1) and inoculated again in sterile beef extract peptone medium at an inoculum volume of 5% to culture until the logarithmic phase. The strains were rinsed with sterile saline to obtain an OD of 1.0. 600 The bacterial suspension is 1.0.

[0071]

[0072] In a sterile environment, the cells were spotted on CMC-Na solid culture medium, inverted in a constant temperature incubator, cultured at 10°C for 48 hours, stained with 1mg / mL Congo red solution, immediately rinsed with sterile water, and then soaked in 1mol / L NaCl for 30 minutes. The hydrolysis zone and community diameter were observed. The analysis results showed that the combination effect of LFS1, ENT1, and VHK1 strains with strain A was not as significant as that of LFS1, ENT1, and VHK1. Figure 5 .

[0073] In a sterile environment, the cells were spotted on aniline blue decolorization medium, placed upside down in a constant temperature incubator, cultured at 25°C for 48 hours, and the decolorization effect was observed. Figure 6 It can be seen that the decolorization effect is best when LFS1 and ENT1 are mixed in equal proportions.

[0074] Comparative Example 2

[0075] Any two of the strains obtained by the above screening were mixed with Bacillus amyloliquefaciens (commercially available with high cellulase production activity, named strain A) to conduct cellulose degradation and lignin degradation experiments, specifically:

[0076] The single bacterial inoculant was a single strain LFS1, ENT1, VHK1, and strain A, which were inoculated into sterile beef extract peptone medium, placed in a constant temperature shaker at 25°C and 180 r / min and cultured for 48 h until the logarithmic phase. The strains were then rinsed with sterile saline to obtain an OD 600The bacterial suspension with an OD of 1.0 is a single bacterial agent. Mix any two of the above-obtained LFS1, ENT1, and VHK1 with strain A in a ratio of 3:1:2, inoculate again in a sterile beef extract peptone medium at an inoculum of 5% and culture to the logarithmic phase. Rinse the strain with sterile saline to obtain an OD of 600 The bacterial suspension is 1.0.

[0077]

[0078] In a sterile environment, the cells were spotted on CMC-Na solid culture medium, inverted in a constant temperature incubator, cultured at 10°C for 48 hours, stained with 1 mg / mL Congo red solution, immediately rinsed with sterile water, and then soaked in 1 mol / L NaCl for 30 minutes. The hydrolysis zone and the diameter of the colony were observed.

[0079] Depend on Figure 7 It can be seen that the strains screened in the above examples, after being mixed with commercially available Bacillus amyloliquefaciens with high cellulose degradation, have a cellulose degradation ability that is not as strong as that of the mixture of LFS1, ENT1, and VHK1.

[0080] In a sterile environment, the cells were spotted on aniline blue decolorization medium, placed upside down in a constant temperature incubator, cultured at 25°C for 48 hours, and the decolorization effect was observed. Figure 7 It can be seen that different bacterial agents all have a degradation effect on lignin after being mixed. Among them, (LFS1, ENT1, VHK1 compound) (LFS1, VHK1, strain A compound) have obvious effects. The decolorization effect of LFS1, ENT1, VHK1 mixed in a ratio of 1:1:1 is the best.

[0081] Example 3: Cow dung maturity test (different temperatures)

[0082] 1. Take fresh cow dung from a farm in Shenyang and mix them evenly in a mass ratio of cow dung: straw (m:m) = 9:1 (the initial properties of cow dung and straw are as follows: pH of cow dung is 8.25, moisture content is 75.36%; pH of straw is 6.78, moisture content is 24.71%), compost them in a foam box, aerate them regularly every day, the volume of the pile is 60L, and the composting cycle is 34 days.

[0083] 2. The compost was carried out in an incubator at 10°C, 20°C, and 30°C. Four treatment groups were set at each temperature: treatment 1 (CK): control treatment; treatment 2 (T1-T3): LFS1, ENT1, and VHK1 in Example 2 were mixed in a ratio of 3:1:2 (T1); 1:3:2 (T2); 1:2:1 (T3) to prepare a mixed microbial agent, which was then loaded on rice husk biochar. The mixed agents were added to the pile respectively, with each microbial agent accounting for 6‰ of the total mass of cow dung; at the same time, the microbial agents of each treatment group were directly added during compost mixing. The temperature index during the composting process was monitored regularly. Fig. 9 Shows the temperature changes during composting at different temperatures. Fig.10 The composting effect of cow dung mixture at different temperatures.

[0084] Depend on Fig. 9 It can be seen that the heating and exposure rate of T1 treatment at different temperatures is better than that of T2 and T3 treatments, and the high temperature (55℃) of T1 treatment at 30℃ lasts for up to 18 days. Fig.10 The composting effect of cow dung mixture at different temperatures shows that under the same treatment, the external temperature is higher, and the texture of the cow dung after fermentation becomes more delicate and soft.

[0085] 3. Perform a quality test on the matured product. Unripe compost products have an inhibitory effect on plant growth. Therefore, the maturity of compost can be evaluated by the growth of plants in a mixture of compost and soil. Seed germination rate experiment is a commonly used biological evaluation method. The maturity and phytotoxicity of compost are judged by comparing the germination of seeds before and after composting. The specific operation is as follows: After the composting process is completed, 10.00 grams of fresh T1, T2, and T3 samples of cow dung fermented at different temperatures in step 2 above are taken and placed in 200 ml conical flasks. According to the actual moisture content of the sample, add an appropriate amount of water at a solid-liquid ratio of 1:10 (mass to volume), then seal the bottle cap and firmly install it on a reciprocating horizontal oscillator. Set the oscillation frequency to 100 times per minute, ensure that the amplitude is not less than 40 mm, and continue to oscillate and extract for 1 hour at 25°C. Afterwards, filter the obtained supernatant through a filter device pre-installed with filter paper, and collect the filtrate for use. Place 1 to 2 sheets of qualitative filter paper in a 9 cm diameter petri dish and evenly place 10 plump carrot seeds of similar size. Add 10 ml of the previously prepared sample filtrate to the petri dish, cover the petri dish lid, and culture in a dark incubator controlled at (25 ± 2) ° C for 48 hours. After the culture is completed, count the number of germinated seeds, and use a vernier caliper to measure the taproot length of each germinated seed one by one, and calculate the germination index based on this. Fig.11 Seed germination index results under different temperature treatments.

[0086] According to the judgment index of compost maturity: if the seed germination index is >50%, it can be considered basically mature, and if the seed germination index reaches 80%-85%, the compost can be considered fully mature. Fig.11 It can be seen that the germination index of cow dung organic fertilizer after treatment with T1, T2 and T3 at different temperatures was higher than 75%, which was basically mature; among them, the seed germination index reached 87.4% under 10℃T1 treatment, 90.1% under 20℃T1 treatment, and 93.6% under 30℃T1 treatment. It can be seen that the composting effect of T1 treatment was completely mature at different temperatures.

[0087] In summary, the decomposition agent provided by the present invention is a biochar microbial composite agent with rapid initiation. The decomposition agent provided by the present invention can accelerate the decomposition process, improve the quality of compost, and will not cause secondary pollution, and is worthy of promotion.

Claims

1. A composting agent, characterized in that: The composting agent is an active ingredient and an adsorption carrier; wherein the active ingredients are Bacillus lysinicus, Klebsiella, and Enterobacter; wherein, Lysinibacillus sp. LFS1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32336; Enterobacter sp. ENT1 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, with the deposit numbers CGMCC No: 32337; Klebsiella VHK1 (Klebsiella sp.) was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on October 24, 2024, and the deposit number is CGMCC No: 32339.

2. The composting agent according to claim 1, characterized in that: The volume mass ratio of the active ingredient to the adsorption carrier is (1-5):

1.

3. The composting agent according to claim 1, characterized in that: The bacterial suspensions of Bacillus lysinicola, Klebsiella and Enterobacter in the active ingredient are in a volume ratio of (1-5):(1-3):(1-2).

4. The composting agent according to any one of claims 1 to 3, characterized in that: The active ingredients are Bacillus lysine, Klebsiella, and Enterobacter, which are placed in LB medium and cultured to the logarithmic growth phase, and the precipitates are collected by centrifugation. Each precipitate is resuspended in sterile saline to OD 600 The bacterial suspension is 0.8-1.2, and then the bacterial suspensions are mixed in a volume ratio of (1-5): (1-3): (1-2), and after mixing, they are fermented in a broth culture medium to obtain a composite fermentation liquid.

5. The composting agent according to claim 4, characterized in that: The bacterial suspensions of lysine bacillus, Klebsiella and Enterobacter in the active ingredients are in a volume ratio of (1-3):(1-2):(1-2).

6. The composting agent according to claim 1 or 2, characterized in that: The adsorption carrier is rice husk biochar.

7. An application of the composting agent according to claim 1, characterized in that: The composting agent is used in preparing organic fertilizer by composting.

8. The use of the composting agent according to claim 7, characterized in that: The composting agent is used in preparing organic fertilizer by composting under low temperature, normal temperature or medium and high temperature conditions.

9. The use of the composting agent according to claim 8, characterized in that: The composting agent is used in starting composting to prepare organic fertilizer under the environmental conditions of low temperature (5°C-20°C), normal temperature (20°C-30°C) or medium-high temperature (above 30°C).

10. The use of the composting agent according to claim 9, characterized in that: The composting agent according to claim 1 is added to agricultural solid waste and composted at low temperature (5°C-20°C), normal temperature (20°C-30°C) or medium-high temperature (above 30°C).

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