CN111620745B - A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste - Google Patents

Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste. By mixing fresh cow dung, lignocellulosic waste, dried distiller's grains and L-alanine fermentation waste liquid, adjust the carbon-nitrogen ratio to 25-30, moisture content 55-65%, 0.1%-0.5% after adding biological inoculants , decomposed and fermented for 6 to 18 days to obtain bio-organic fertilizer. The present invention utilizes dried distiller's grains and its enzymatic hydrolyzate to provide nutrients and growth-promoting factors required in the fermentation process of functional microorganisms, increases the number of effective viable bacteria in the biological inoculants, and reduces the nutritional difference in the decomposed fermentation environment; L-alanine is used for fermentation The bacterial protein and L-alanine components in the waste liquid can accelerate the germination of spores. The two effectively supplement various amino acids, vitamins and calcium and phosphorus mineral components, improve the nutritional quality and functional potency of bio-organic fertilizers, and help increase crop production and income and improve quality.

Description

A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste

technical field

The invention belongs to the field of biotechnology, and in particular relates to a method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste.

Background technique

With the rapid development of modern agriculture in my country, a large number of agricultural wastes such as straw and livestock manure are generated, and the corresponding environmental pollution problems are gradually intensifying, which is not conducive to the sustainable development of society and economy. Among them, the annual output of agricultural straw lignocellulosic waste in my country exceeds 900 million tons, and the annual output of livestock and poultry manure reaches 3.8 billion tons. The potential for resource utilization is huge. Lignocellulosic waste is rich in organic matter, cellulose, crude protein, crude fat, and nutrients such as nitrogen, phosphorus, potassium, calcium, magnesium, and sulfur. The nutrient content of 1 ton of dry straw is equivalent to 50-60 kg of fertilizer. In addition, livestock and poultry manure such as cow dung also contains various nutrients such as nitrogen, phosphorus and potassium necessary for crops, and the nutrient content of 1 ton of manure is equivalent to 20-30 kg of chemical fertilizer. According to the circular economy concept of "reduction, reuse, and resource utilization" in my country, using these agricultural waste resources to produce green bio-organic fertilizers, and supporting the promotion and use of commercial organic fertilizers will effectively promote modern agriculture to achieve "waste recycling, resource Efficient utilization, clean and controllable production” green sustainable development goals, and promote the development of modern agricultural circular economy.

The decomposition and fermentation of waste biomass into fertilizer is to promote the decomposition of organic materials such as livestock and poultry manure into fertilizer nutrients required by crops through the catabolism of microorganisms, and to kill pathogenic microorganisms such as bacteria and fungi, as well as the biotransformation of their spores and eggs. The addition of compound inoculants containing a variety of functional microorganisms can significantly improve the decomposing process and the quality of organic fertilizers. At present, the functional microorganisms widely used in decomposing fermentation are mainly derived from Bacillus, Trichoderma, Aspergillus and Saccharomyces. Among them, Bacillus, Trichoderma and Aspergillus microorganisms are added in the form of spores. In the actual decomposed fermentation environment, macromolecular organic substances need to be degraded into small molecular substances, and nutrients such as carbohydrate carbon sources, amino acids and growth-promoting factors are insufficient. It is conducive to the germination and growth of spores, resulting in low degradation efficiency of organic matter, insufficient decomposing, long fermentation time, and incomplete organic fertilizer nutrients, which need to be solved urgently.

Taking cow dung decomposing fermentation as an example, most of the current research reports focus on optimizing the selection of auxiliary materials, material particle size, water content of the pile, carbon-nitrogen ratio, functional microorganism screening, and preparation of bacterial agents. Zhang Yufeng et al. ^[1] used the aerobic fermentation method after the addition of biological inoculants to explore the optimal choices of corn stalk, furfural residue and fungus residue as the auxiliary materials for cow dung decomposed fermentation, and found that the combination of cow dung + furfural residue + straw heats up faster , the high temperature maintained for a long time, and under the combination of cow dung + straw + urea, the total nutrient content of the decomposed organic fertilizer reached the highest 5.04%, and the decomposed fermentation time was 23 days. Zhang He et al. ^[2] studied and reported the effect of different carbon-nitrogen ratios on the aerobic decomposing process of cow dung, indicating that in actual production, the carbon-nitrogen ratio of cow dung and straw mixture between 25 and 30: 1 is beneficial The heap is decomposed and nutrients are maintained. Han Xianglong et al. ^[3] also studied and reported the effect of different carbon-nitrogen ratios on the decomposing process of tobacco stems and cow dung. The content of other nutrients is the highest, and the comprehensive effect of decomposed fermentation is the best. Zhang Yufeng et al ^[4] explored the screening of aerobic fermentation inoculants mixed with cow dung and corn stalk. The decomposing effect of manure and straw is remarkable, and the decomposing fermentation time is about 23 days. Tian Zhihui et al. ^[5] studied the initial conditions of aerobic decomposing fermentation of cow dung and straw. The results showed that adjusting the moisture content of the heap to about 60%, the carbon to nitrogen ratio to about 30: 1, and the size of the pulverized wheat straw ≤ 1 cm, the best effect was achieved. good. The patent CN110668861A discloses a method for producing organic fertilizer by utilizing the decomposing and fermenting of chlorosis and cow dung. The raw materials include 30% to 70% of fresh chlorosis, 12% to 52% of cow dung, 5% of straw, 10% of sawdust Sawdust and 3% waste biomass carbon, the moisture content is adjusted to 60% to 70%, and the carbon to nitrogen ratio is 25 to 30:1. The compound bacterial agent used is composed of Bacillus subtilis, Bacillus licheniformis, Saccharomyces cerevisiae and Trichoderma viride spore suspension , The decomposing fermentation process is divided into two stages, the first is aerobic fermentation for 9 to 12 days, followed by the secondary fermentation for 21 to 27 days, but the decomposing fermentation time is slightly longer.

The above statement of the background technology is only for the convenience of in-depth understanding of the technical solutions of the present invention (technical means used, technical problems solved and technical effects produced, etc.) It constitutes the prior art known to those skilled in the art.

SUMMARY OF THE INVENTION

In view of the problems existing in the prior art, the purpose of the present invention is to provide a method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural wastes, aiming to solve the problem that in the existing decomposing fermentation technology, the nutrients and promoting Insufficient growth factors lead to problems such as slow germination and growth and proliferation of microbial spores, low degradation efficiency of raw materials, insufficient decomposing, long fermentation time, and incomplete organic fertilizer nutrients. At the same time, it also provides a new way for the full utilization of bacterial protein and residual L-alanine in the L-alanine fermentation waste liquid, and reduces the treatment cost of the L-alanine fermentation waste liquid. The invention utilizes the dried distiller's grains and the L-alanine fermentation waste liquid to improve the production efficiency, nutritional quality and functional potency of the biological organic fertilizer.

In order to solve the above-mentioned technical problems, the present invention adopts the following technical solutions:

The invention provides a method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste, comprising the following steps:

S1: Mix the fresh cow dung, lignocellulosic waste and dried distiller's grains to form a pile, and adjust the carbon-nitrogen ratio of the pile to 25-30;

S2: adding the L-alanine fermentation waste liquid to the pile body obtained in step S1, stirring evenly, and controlling the pH of the pile body to which the L-alanine fermentation waste liquid is added is 7-8, and the water content is 55-65%;

S3: adding the biological inoculum to the pile of the L-alanine fermentation waste liquid obtained in step S2, and stirring evenly, the usage amount of the biological inoculum is the total mass of the pile with the addition of the L-alanine fermentation waste liquid 0.1%-0.5% of ;

S4: use 100kg scale static decomposing fermentation for 15-18 days, or 1-10 ton scale bioreactor fermentation for 6-8 days, and then add bio-control probiotics to obtain bio-organic fertilizer, and the number of effective viable bacteria in the fertilizer is not less than 20 million/g.

In the above technical scheme, further, the lignocellulosic waste includes one or more of corn stalk, wheat stalk, rice stalk, sorghum stalk, rice straw stalk, wheat bran, chaff, corn cob and rice bran, mixed together. Before crushing to 1-2cm.

In the above technical scheme, the biocontrol probiotics are selected from Bacillus subtilis, Bacillus firmus, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, Bacillus jelly-like and Chain yellow One or more of the molds.

In the above-mentioned technical scheme, further, the L-alanine fermentation waste liquid is fermented by Escherichia coli, Escherichia coli or Pseudomonas, and produced after extracting L-alanine, comprising bacterial protein and residues. L-alanine liquid.

In the above technical scheme, further, the quality of the dried distiller's grains is 5%-10% of the quality of the fresh cow dung.

In the above technical scheme, further, the biological inoculum is prepared by directly mixing Bacillus subtilis fermentation liquid, Aspergillus niger fermentation liquid, Trichoderma reesei fermentation liquid and Saccharomyces cerevisiae fermentation liquid, and the number of effective viable bacteria in the biological inoculum is ≥ 6.6×10 ⁹ CFU/mL. The number of effective viable bacteria in the fermentation broth of Bacillus subtilis ≥1.6×10 ¹⁰ CFU/mL, the number of viable viable bacteria in the fermentation broth of Aspergillus niger ≥3.2×10 ⁹ CFU/mL, and the number of viable viable bacteria in the fermentation broth of Trichoderma reesei ≥3.6 ×10 ⁹ CFU/mL, the number of effective viable bacteria in the fermentation broth of Saccharomyces cerevisiae ≥ 4.8 × 10 ⁹ CFU/mL.

In the above-mentioned technical scheme, further, the medium composition for preparing Bacillus subtilis, Aspergillus niger, Trichoderma reesei and Saccharomyces cerevisiae fermentation liquid comprises:

(1) Bacillus subtilis medium: glucose 30-70g/L, peptone 5-15g/L, beef extract 10-30 g/L, yeast powder 5-15g/L, calcium carbonate 5-15g/L, the rest are Dried distiller's grains enzymolysis solution, pH7-8;

(2) Aspergillus niger and Trichoderma reesei culture medium: glucose 30-60g/L, malt extract powder 5-15g/L, (NH ₄ ) ₂ SO ₄ 5-15g/L, calcium carbonate 5-15g/L, The rest are dry distiller's grains enzymolysis solution, pH7-8;

(3) Saccharomyces cerevisiae culture medium: glucose 60-120g/L, yeast powder 5-15g/L, peptone 20-40 g/L, calcium carbonate 5-15g/L, the rest are dry distiller's grains enzymolysis solution, pH6-7 .

Wherein, the dry distiller's grains enzymolysis solution is prepared by the following method: the dry distiller's grains are evenly mixed with water at a mass ratio of 5%-10%, adjusted to pH 8-11 with ammonia solution, water bath at 80°C for 60 minutes, under the condition of 50°C, Add alkaline protease for enzymatic hydrolysis at 200-600 U/g-dry distiller's grains for 24 hours, centrifuge, remove the precipitation, and the supernatant is the dry distiller's grains enzymatic hydrolysis solution.

In the above technical solution, further, during the static decomposing fermentation process of the 100kg scale, the heap is not turned over before the temperature rises to 55°C, and after the temperature rises to 65°C, the heap is turned every day to keep the temperature not higher than 70°C; Drop to 50°C, and turn the heap once every two days. Mature organic fertilizer is obtained after 15-18 days of decomposing and fermentation, and then adding biocontrol probiotics such as Bacillus subtilis, Bacillus firmus, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, jelly-like spores One or more of bacillus and Streptomyces flavonoids to obtain bio-organic fertilizer.

In the above technical scheme, further, in the fermentation process of the 1-10 ton scale bioreactor, the initial fermentation temperature is controlled to be 30°C, and the control is released after the temperature rise; the ventilation frequency is 2-3 times/day; 3 times, the rotation speed is 50-150rpm; after 6-8 days of fermentation, the machine is dewatered or air-dried to obtain mature organic fertilizer; then add biocontrol probiotics such as Bacillus subtilis, Bacillus firmus, Paenibacillus polymyxa, lichen One or more of Bacillus megaterium, Bacillus megaterium, Bacillus amyloliquefaciens, Bacillus jelly-like and Streptomyces flavonoids can be obtained as biological organic fertilizer.

The bio-organic fertilizer produced by the above technical solution has a slightly fermented fragrance, and all indicators have reached or exceeded the national industry standard for organic fertilizers (NY 525-2012) and the agricultural industry bio-organic fertilizer standard of the People's Republic of China (NY 884-2012).

Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention utilizes the dried distiller's grains to provide functional microorganisms with nutrients and growth-promoting factors required for fermentation and decomposing fermentation, thereby reducing nutritional differences in the growth microenvironment; on the other hand, effectively utilizing L -Bacterial protein and L-alanine components in the alanine fermentation waste liquid, accelerate the germination of spore-producing microorganism spores, and replace other water sources with fermentation waste liquid, saving water; Thorough, fast reproduction of functional microorganisms and better control of fermentation conditions (such as aeration, stirring, nutrient transfer, etc.) The fermentation waste liquid also effectively supplemented more than ten kinds of amino acids (alanine, arginine, aspartic acid, cysteine, glutamic acid and leucine, etc.), vitamins and calcium and phosphorus minerals, which improved the The organic matter and total nutrient content of organic fertilizers improve the nutritional quality and functional potency, which is conducive to increasing crop production and income.

Detailed ways

The present invention will be further described below in conjunction with the examples. It should be understood that the scope of protection claimed in the present invention is not limited by the specific embodiments. The specific examples provided in the present invention are only used as examples to further illustrate the present invention. The description in the specification of the present application can easily modify the specific embodiments of the present invention or perform equivalent replacements to some technical features, and these improvements and replacements without creative work should also fall within the protection scope of the appended claims of the present invention. Materials of the same or similar type, model, quality, nature or function as the following reagents and instruments can be used in the practice of the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

The used bacterial strains, reagents and raw material sources in the following examples of the present invention are as follows:

Bacillus subtilis ACCC 19742, Aspergillus niger ACCC 32589, and Saccharomyces cerevisiae ACCC 21162 were purchased from the China Agricultural Microorganism Culture Collection and Management Center, and the used Trichoderma reesei Rut C30 (NRRL 11460) was donated to the American Agricultural Research Culture Collection.

Biochemical reagents such as glucose, peptone, beef extract, yeast powder, alkaline protease were purchased from Shenggong Bioengineering (Shanghai) Co., Ltd., fresh cow dung, dried distiller's grains, straw, wheat bran, rice husk and L-alanine fermentation waste Liquid and other raw materials were provided by Dalian Jinweide Biotechnology Co., Ltd.

Example 1:

A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste, the specific steps for preparing biological inoculants are as follows:

1. Prepare dry distiller's grains enzymolysis solution:

Mix the dried distiller's grains with tap water at a mass ratio of 10%, adjust the pH to 9 with an ammonia solution, take a water bath at 80°C for 60 minutes, add 400U/g-dry distiller's grains at 50°C for enzymatic hydrolysis with alkaline protease for 24 hours, and then centrifuge. That is, the dry distiller's grains enzymolysis solution is obtained.

2. Strain activation

Bacillus subtilis was spread on LB (Luria-Bertani Agar) medium and cultured at 30°C for 20 hours to obtain Bacillus subtilis colonies; Aspergillus niger and Trichoderma reesei were spread on PDA (Potato Dextrose Agar) medium, respectively, at 30°C After culturing for 96 hours, Aspergillus niger and Trichoderma reesei spores were obtained; Saccharomyces cerevisiae was spread on YPD (Yeast Extract Peptone Dextrose Agar) medium and cultured at 30° C. for 20 hours to obtain Saccharomyces cerevisiae colonies.

3. Fermentation culture

(1) Pick a single colony of Bacillus subtilis after activation, inoculate it in a liquid fermentation medium, cultivate at 150 rpm at a constant temperature of 30° C. for 24 hours to obtain a Bacillus subtilis fermentation broth, and the number of effective viable bacteria is 1.6×10 ¹⁰ CFU/mL Among them, the fermentation medium is composed of: glucose 60g/L, peptone 15g/L, beef extract 30g/L, yeast powder 15g/L, calcium carbonate 15g/L, and the rest are dry distiller's grains enzymolysis solution, pH 7.2.

(2) The activated Aspergillus niger spores were collected with sterile water, inoculated into a liquid fermentation medium, cultured at a constant temperature of 150 rpm and 30° C. for 72 hours to obtain Aspergillus niger fermentation broth, and the number of effective viable bacteria was 3.2×10 ⁹ CFU/ Among them, the fermentation medium is composed of: glucose 50g/L, malt extract powder 15g/L, (NH ₄ ) ₂ SO ₄ 15g/L, calcium carbonate 15g/L, and the rest are dry distiller's grains enzymolysis solution, pH 7.2 .

(3) The activated Trichoderma reesei spores were collected with sterile water, inoculated into a liquid fermentation medium, cultured at 150 rpm at a constant temperature of 30° C. for 72 hours to obtain a Trichoderma reesei fermentation broth, and the number of effective viable bacteria was 3.6× 10 ⁹ CFU/mL; wherein, the fermentation medium consists of: glucose 50g/L, malt extract powder 15g/L, (NH ₄ ) ₂ SO ₄ 15g/L, calcium carbonate 15g/L, and the rest are dry distiller's grains enzymolysis solution , pH 7.2.

(4) Picking a single colony of Saccharomyces cerevisiae after activation, inoculating it into a liquid fermentation medium, culturing at 150 rpm at a constant temperature of 30 °C for 24 hours, to obtain a fermentation broth of Saccharomyces cerevisiae, and the effective viable count is 4.8×10 ⁹ CFU/mL; Among them, the fermentation medium is composed of: glucose 100g/L, yeast powder 15g/L, peptone 40g/L, calcium carbonate 15g/L, and the rest are dry distiller's grains enzymolysis solution, pH 6.5.

4. Fermentation broth mixing

The above-mentioned Bacillus subtilis fermentation broth, Aspergillus niger fermentation broth, Trichoderma reesei fermentation broth and Saccharomyces cerevisiae fermentation broth were evenly mixed to obtain a biological inoculum, and the number of effective viable bacteria in the inoculum was 6.6×10 ⁹ CFU/mL.

Control experiment:

1. Strain activation

Bacillus subtilis was spread on LB (Luria-Bertani Agar) medium and cultured at 30°C for 20 hours to obtain Bacillus subtilis colonies; Aspergillus niger and Trichoderma reesei were spread on PDA (Potato Dextrose Agar) medium, respectively, at 30°C After culturing for 96 hours, Aspergillus niger and Trichoderma reesei spores were obtained; Saccharomyces cerevisiae was spread on YPD (Yeast Extract Peptone Dextrose Agar) medium and cultured at 30° C. for 20 hours to obtain Saccharomyces cerevisiae colonies.

2. Fermentation culture

(1) Picking a single colony of Bacillus subtilis after activation, inoculating it into a liquid fermentation medium, culturing at 150 rpm at a constant temperature of 30° C. for 24 hours to obtain a Bacillus subtilis fermentation broth, and the number of effective viable bacteria is 7.5×10 ⁹ CFU/mL Among them, the fermentation medium consists of: glucose 60g/L, peptone 15g/L, beef extract 30g/L, yeast powder 15g/L, calcium carbonate 15g/L, the rest is tap water, pH 7.2.

(2) The activated Aspergillus niger spores were collected with sterile water, inoculated into a liquid fermentation medium, cultured at a constant temperature of 150 rpm and 30° C. for 72 hours to obtain Aspergillus niger fermentation broth, and the number of effective viable bacteria was 4.2×10 ⁸ CFU/ Among them, the fermentation medium consists of: glucose 50g/L, malt extract powder 15g/L, (NH ₄ ) ₂ SO ₄ 15g/L, calcium carbonate 15g/L, and the rest are tap water, pH 7.2.

(3) The activated Trichoderma reesei spores are collected with sterile water, inoculated in a liquid fermentation medium, cultured at 150 rpm at a constant temperature of 30° C. for 72 hours to obtain a Trichoderma reesei fermentation broth, and the number of effective viable bacteria is 4.5× 10 ⁸ CFU/mL; wherein, the fermentation medium is composed of: glucose 50g/L, malt extract powder 15g/L, (NH ₄ ) ₂ SO ₄ 15g/L, calcium carbonate 15g/L, the rest are tap water, pH7.2 .

(4) Picking a single colony of Saccharomyces cerevisiae after activation, inoculating it into a liquid fermentation medium, culturing at 150 rpm at a constant temperature of 30° C. for 24 hours to obtain a fermentation broth of Saccharomyces cerevisiae, and the effective viable count is 1.0×10 ⁹ CFU/mL; Among them, the fermentation medium was composed of: glucose 100g/L, yeast powder 15g/L, peptone 40g/L, calcium carbonate 15g/L, and the rest was tap water, pH 6.5.

3. Fermentation broth mixing

The above-mentioned Bacillus subtilis fermentation broth, Aspergillus niger fermentation broth, Trichoderma reesei fermentation broth and Saccharomyces cerevisiae fermentation broth were evenly mixed to obtain a biological inoculum with an effective viable bacterial count of 2.2×10 ⁹ CFU/mL.

The invention utilizes the abundant nutrients and growth-promoting factors in the dried distiller's grains enzymatic hydrolyzate to carry out functional microorganism fermentation and cultivation, and compared with the Bacillus subtilis, Aspergillus niger, Trichoderma viride and Saccharomyces cerevisiae fermentation broth prepared by conventional methods, the number of effective viable bacteria is greatly increased , the number of effective viable bacteria in the prepared biological inoculum was increased by 3 times.

Example 2:

A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste, the specific decomposing and fermentation steps are as follows:

1. Mixed heap

Mix fresh cow dung, crushed straw and dry distiller's grains, adjust the carbon-nitrogen ratio to 30, wherein, the amount of dry distiller's grains is 10% of the quality of fresh cow dung; add L-alanine fermentation waste liquid, stir evenly, pH 7.8, Control the moisture content to 65%; then add 0.5% of the total mass of the heap after adjusting the moisture content, add the biological inoculum prepared in Example 1 (the number of effective viable bacteria is 6.6×10 ⁹ CFU/mL), mix evenly, and the total heap The mass is close to 100kg.

2. Decomposed and fermented

The 100kg scale static decomposing fermentation is adopted. When the temperature rises to 55°C, the heap is not turned; when the temperature rises to 65°C, the heap is turned every day to keep the temperature not higher than 70°C; when the temperature drops to 50°C, the heap is turned over for two days. Once; when the temperature drops to 35 ℃, stop turning the heap, decompose and ferment for 15 days to obtain mature organic fertilizer, and then add bio-control probiotic Bacillus firmus to obtain bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 050 million/g, the mass fraction of organic matter is 62.5%, the mass fraction of total nutrients is 9.2%, the mass fraction of water is 27.6%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 2/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 7.8, all indicators have reached or exceeded the national industry standard for organic fertilizers (NY525-2012) and the agricultural industry of the People's Republic of China Standard for Bio-Organic Fertilizers (NY 884-2012). After the application of the biological organic fertilizer, it can quickly inhibit and kill the rhizosphere parasitic root-knot nematodes, cyst nematodes, golden nematodes, heterodermic nematodes and other harmful organisms, eggs and larvae, and at the same time provide crops with abundant organic matter, nitrogen, phosphorus and potassium. , amino acids, vitamins and minerals and other nutrients.

Control experiment:

1. Mixed heap

Mix fresh cow dung and crushed straw, add tap water, adjust the carbon-nitrogen ratio to 30, pH 7.8, and control the moisture content to 65%; add the biological bacteria prepared in Example 1 according to 0.5% of the total mass of the pile after adjusting the moisture content agent (the number of effective viable bacteria is 2.2×10 ⁹ CFU/mL), mix well, and the total mass of the pile is close to 100kg.

2. Decomposed and fermented

The 100kg scale static decomposing fermentation is adopted. When the temperature rises to 55°C, the heap is not turned; when the temperature rises to 65°C, the heap is turned every day to keep the temperature not higher than 70°C; when the temperature drops to 50°C, the heap is turned over for two days. Once; when the temperature drops to 35°C, stop turning the heap, decompose and ferment for 30 days to obtain mature organic fertilizer, and then add bio-control probiotic Bacillus firmus to obtain bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 50 million/g, the mass fraction of organic matter is 50.0%, the mass fraction of total nutrients is 6.6%, the mass fraction of water is 28.7%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 5/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 8.0, all indicators also meet or exceed the national industry standard for organic fertilizers (NY525-2012) and the agricultural industry of the People's Republic of China. Organic Fertilizer Standard (NY 884-2012). However, in Example 2, the decomposing fermentation time was only 15 days, the production efficiency of the bio-organic fertilizer was doubled, and the nutritional quality and functional titer of the bio-organic fertilizer were also greatly improved.

Example 3:

A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste, the specific decomposing and fermentation steps are as follows:

1. Mixed heap

Mix fresh cow dung, pulverized wheat bran and dry distiller's grains, adjust the carbon-nitrogen ratio to 25, wherein the amount of dry distiller's grains is 5% of the quality of fresh cow dung; add L-alanine fermentation waste liquid, stir evenly, pH 7.8 , control the moisture content to 55%; then add 0.1% of the total mass of the pile after adjusting the moisture content, add the biological inoculum prepared in Example 1 (the number of effective viable bacteria is 6.6×109CFU/mL), mix evenly, and the total mass of the pile is close to 100kg.

2. Decomposed and fermented

The 100kg scale static decomposing fermentation is adopted. When the temperature rises to 55°C, the heap is not turned; when the temperature rises to 65°C, the heap is turned every day to keep the temperature not higher than 70°C; when the temperature drops to 50°C, the heap is turned over for two days. Once; when the temperature drops to 35°C, stop turning the heap, and ferment for 18 days to obtain mature organic fertilizer, and then add bio-control probiotic Streptomyces flavus to obtain bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.3 billion/g, the mass fraction of organic matter is 60.4%, the mass fraction of total nutrients is 8.4%, the mass fraction of water is 25.9%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 2/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 7.5, all indicators have reached or exceeded the national industry standard for organic fertilizers (NY525-2012) and the agricultural industry of the People's Republic of China Standard for Bio-Organic Fertilizers (NY 884-2012). After the application of the bio-organic fertilizer, it can improve soil fertility, inhibit the reproduction of pathogenic microorganisms, promote the rooting, germination and maturity of crops, increase production and income, and improve the quality of agricultural products. Substances and other nutrients.

Control experiment:

1. Mixed heap

Mix fresh cow dung and pulverized wheat bran, add tap water, adjust the carbon-nitrogen ratio to 25, pH 7.8, and control the moisture content to 55%; add the biological material prepared in Example 1 by 0.1% of the total mass of the heap after adjusting the moisture content. Bacterial agent (the number of effective viable bacteria is 2.2×109CFU/mL), mix well, and the total mass of the pile is close to 100kg.

2. Decomposed and fermented

The 100kg scale static decomposing fermentation is adopted. When the temperature rises to 55°C, the heap is not turned; when the temperature rises to 65°C, the heap is turned every day to keep the temperature not higher than 70°C; when the temperature drops to 50°C, the heap is turned over for two days. Once; when the temperature drops to 35 ℃, stop turning the heap, decompose and ferment for 35 days to obtain mature organic fertilizer, and then add bio-control probiotic Bacillus firmus to obtain bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.2 billion/g, the mass fraction of organic matter is 48.8%, the mass fraction of total nutrients is 6.1%, the mass fraction of water is 23.7%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 5/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 7.6, all indicators also meet or exceed the national industry standard for organic fertilizers (NY525-2012) and the agricultural industry of the People's Republic of China. Organic Fertilizer Standard (NY 884-2012). However, in Example 3, the decomposing fermentation time was only 18 days, the production efficiency of the bio-organic fertilizer was nearly doubled, and the nutritional quality and functional titer were also greatly improved.

Example 4:

A method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste, the specific decomposing and fermentation steps are as follows:

1. Mixed heap

Mix fresh cow dung, pulverized rice husks and dry distiller's grains, adjust the carbon-nitrogen ratio to 28, wherein the amount of dry distiller's grains is 8% of the quality of fresh cow dung; add L-alanine fermentation waste liquid, stir evenly, pH 7.8 , control the moisture content to 60%; then add the biological inoculum prepared in Example 1 (the number of effective viable bacteria is 6.6×10 ⁹ CFU/mL) according to 0.3% of the total mass of the pile after adjusting the moisture content, mix evenly, and the total mass close to 100kg.

2. Decomposed and fermented

A 5-ton scale bioreactor was used for fermentation, and the initial fermentation temperature was controlled to be 30 °C, and the control was released after the temperature was raised; the ventilation frequency was 3 times/day; the stirring was 3 times a day, and the rotation speed was 100 rpm; After the temperature drops to 35°C, the machine removes water or air-dries, and the mature organic fertilizer is obtained in 7 days; then the bio-control probiotic Paenibacillus polymyxa is added to obtain the bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.5 billion/g, the mass fraction of organic matter is 58.6%, the mass fraction of total nutrients is 8.8%, the mass fraction of water is 29.3%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 2/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 7.9, all indicators also meet or exceed the national industry standard for organic fertilizers (NY 525-2012) and the agricultural industry of the People's Republic of China Standard for Bio-Organic Fertilizers (NY 884-2012). The bio-organic fertilizer can effectively prevent and control bacterial and fungal soil-borne diseases of plants, such as plant bacterial wilt and fusarium wilt, significantly reduce bacterial and fungal diseases on plant leaves, and has an obvious effect of promoting growth and increasing yield of crops. At the same time, it provides rich nutrients such as organic matter, nitrogen, phosphorus and potassium, amino acids, vitamins and minerals for crops.

Control experiment:

1. Mixed heap

Mix fresh cow dung and crushed rice husks, add tap water, adjust the carbon-nitrogen ratio to 28, pH 7.8, and control the moisture content to 60%; add the biomass prepared in Example 1 by 0.3% of the total mass of the heap after adjusting the moisture content. Bacterial agent (the number of effective viable bacteria is 2.2×10 ⁹ CFU/mL), mix well, and the total mass of the pile is close to 100kg.

2. Decomposed and fermented

A 5-ton scale bioreactor was used for fermentation, and the initial temperature was controlled at 30 °C, and the control was released after the temperature was raised; the ventilation frequency was 3 times/day; the stirring was 3 times a day, and the rotation speed was 100 rpm; After the temperature drops to 35°C, the machine removes water or air-dries, and mature organic fertilizer can be obtained within 12 days; then bio-control probiotic Paenibacillus polymyxa is added to obtain bio-organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 50 million/g, the mass fraction of organic matter is 50.1%, the mass fraction of total nutrients is 6.5%, the mass fraction of water is 29.5%, the death rate of Ascaris eggs reaches 100%, and the fecal coliform bacteria The number of clusters is less than 2/g, total arsenic, mercury, lead, cadmium and chromium are not detected, pH 7.7, all indicators also meet or exceed the national industry standard for organic fertilizers (NY 525-2012) and the agricultural industry of the People's Republic of China Standard for Bio-Organic Fertilizers (NY 884-2012). However, in Example 4, the decomposing fermentation time was only 7 days, the production efficiency of the bio-organic fertilizer was increased by more than 70%, and the nutritional quality and functional titer were also greatly improved.

For any person skilled in the art, without departing from the scope of the technical solution of the present invention, many possible changes and modifications can be made to the technical solution of the present invention by using the technical content disclosed above, or modified into equivalents of equivalent changes Example. Therefore, any simple modifications, equivalent changes and modifications made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solutions of the present invention should still fall within the protection scope of the technical solutions of the present invention.

references

1. Zhang Yufeng, Tian Shenzhong, Bian Wenfan, Lin Haitao, Wang Jiangtao, Dong Liang, Liu Zhaohui, Guo Honghai, Ren Qingju. The effect of organic materials on the aerobic composting process of cow dung. Shandong Agricultural Science, 2019,51(05):76- 82.

2. Zhang He, Li Mengchan, Yang Huizhen, Wang Youling, Lu Yongli, Zhang Chunhong, Qiu Huizhen. Effects of different carbon and nitrogen ratios on the decomposing process of aerobic composting of cow manure. Journal of Gansu Agricultural University, 2019, 54(01): 60-67.

3. Han Xianglong, Wu Wei, Zhao Pengbo, Wei Chengcai, Yuan Shuai, Li Sitong, Mao Kailun, Niu Gaili, Zhang Lixin. Effects of different carbon and nitrogen ratios on the composting process of tobacco stems and cow dung. Jiangsu Agricultural Science, 2019, 47( 16):303-307.

4. Zhang Yufeng, Tian Shenzhong, Bian Wenfan, Guo Honghai, Gong Zhiyuan, Liu Zhaohui, Li Ruiqin, Chen Jianqiu, Luo Jiafa. Screening of aerobic fermentation inoculants in mixed composting of cow dung and corn stalk. Soil and Fertilizer in China, 2019, 172-178 .

5. Tian Zhihui, Wang Yani, Zou Chao, Lu Jundang. Study on the initial conditions of aerobic fermentation and composting of cow dung and straw. China Soil and Fertilizer, 2018(06): 192-197.

Claims (5)

1. a method for utilizing biological inoculum to degrade agricultural waste to produce biological organic fertilizer, is characterized in that, comprises the following steps: S1: Mix the fresh cow dung, lignocellulosic waste and dried distiller's grains to form a pile, and adjust the carbon-nitrogen ratio of the pile to 25-30; S2: adding the L-alanine fermentation waste liquid to the pile obtained in step S1, stirring evenly, and controlling the pH of the pile to which the L-alanine fermentation waste liquid is added is 6-8, and the water content is 55-65%; S3: adding the biological inoculum to the pile of the L-alanine fermentation waste liquid obtained in step S2, and stirring evenly, the usage amount of the biological inoculum is the total mass of the pile with the addition of the L-alanine fermentation waste liquid 0.1%-0.5% of ; S4: Use 100 kg scale static decomposing fermentation for 15-18 days, or 1-10 ton scale bioreactor fermentation for 6-8 days, and then supplement bio-control probiotics to obtain bio-organic fertilizer, and the number of effective viable bacteria in the fertilizer is not low at 0.2 billion/g; The L-alanine fermentation waste liquid is a liquid containing bacterial protein and residual L-alanine produced after extraction of L-alanine by fermentation of Escherichia coli, Escherichia coli or Pseudomonas; The quality of the dried distiller's grains is 5%-10% of the fresh cow dung quality; The biological inoculum is prepared by directly mixing Bacillus subtilis fermentation liquid, Aspergillus niger fermentation liquid, Trichoderma reesei fermentation liquid and Saccharomyces cerevisiae fermentation liquid, and the number of effective viable bacteria in the biological inoculum is ≥6.6×10 ⁹ CFU/ mL; wherein, the number of effective viable bacteria in the Bacillus subtilis fermentation broth ≥ 1.6×10 ¹⁰ CFU/mL, the number of effective viable bacteria in the Aspergillus niger fermentation broth ≥ 3.2×10 ⁹ CFU/mL, and the Trichoderma reesei fermentation broth The number of effective viable bacteria ≥ 3.6×10 ⁹ CFU/mL, and the number of effective viable bacteria in the fermentation broth of Saccharomyces cerevisiae ≥ 4.8×10 ⁹ CFU/mL; The preparation of the Bacillus subtilis fermentation liquid, the Aspergillus niger fermentation liquid, the Trichoderma reesei fermentation liquid and the Saccharomyces cerevisiae fermentation liquid are sequentially activated and cultured and fermented, and the composition of the fermentation medium for each strain includes: (1) Bacillus subtilis: glucose 30-70 g/L, peptone 5-15 g/L, beef extract 10-30 g/L, yeast powder 5-15 g/L, calcium carbonate 5-15 g/L, the rest It is dry distiller's grains enzymolysis solution, pH7-8; (2) Aspergillus niger and Trichoderma reesei: glucose 30-60 g/L, malt extract powder 5-15 g/L, (NH ₄ ) ₂ SO ₄ 5-15 g/L, calcium carbonate 5-15 g/L L, the rest are dry distiller's grains enzymolysis solution, pH7-8; (3) Saccharomyces cerevisiae: glucose 60-120 g/L, yeast powder 5-15 g/L, peptone 20-40 g/L, calcium carbonate 5-15 g/L, the rest are dry distiller's grains enzymatic hydrolysate, pH 6- 7; The dry distiller's grains enzymolysis solution is prepared by the following method: the dry distiller's grains are mixed evenly with water at a mass ratio of 5%-10%, adjusted to pH 8-11 with an aqueous ammonia solution, water bath at 80°C for 60 minutes, and under the condition of 50°C, press 200 ~600 U/g-Dry distiller's grains were added with alkaline protease for enzymatic hydrolysis for 24 hours, centrifuged to remove the precipitation, and the supernatant was the dry distiller's grains enzymatic hydrolysis solution. 2. The method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural waste according to claim 1, wherein the lignocellulosic waste comprises corn stalks, wheat stalks, rice stalks, sorghum stalks, and rice straw stalks , wheat bran, rice husk, corn cob and rice bran or one or more of them, crushed to 1-2 cm before mixing. 3. the method that utilizes biological inoculum to degrade agricultural waste according to claim 1 and 2 to produce biological organic fertilizer, it is characterized in that, described biocontrol probiotic is selected from Bacillus subtilis, Bacillus firmus, polymyxa One or more of Bacillus, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, Bacillus jelly-like and Streptomyces flavis. 4. the method that utilizes biological inoculum to degrade agricultural waste to produce biological organic fertilizer according to claim 1 and 2, it is characterized in that, in described 100 kg scale static decomposing fermentation process, temperature rises to 55 ℃ before not turning the heap , after the temperature rises to 65 °C, turn the heap every day to keep the temperature not higher than 70 °C; when the temperature drops to 50 °C, turn the heap once every two days; 15-18 days of decomposing and fermentation to obtain mature organic fertilizer, and then add Bacillus subtilis , one or more of Bacillus firmus, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, Bacillus jelly-like and Streptomyces lividans to obtain biological organic fertilizer. 5. The method for producing bio-organic fertilizer by utilizing biological inoculants to degrade agricultural wastes according to claim 1 or 2, wherein in the fermentation process of the 1-10 ton scale bioreactor, the initial temperature is controlled to be 30°C , release the control after heating up; aeration frequency 2-3 times/day; stirring 2-3 times a day, rotating speed 50-150 rpm; after 6-8 days of decomposing fermentation, then dewatering or air-drying by machine, that is, mature organic fertilizer; Then add one or more of Bacillus subtilis, Bacillus firmus, Bacillus polymyxa, Bacillus licheniformis, Bacillus megaterium, Bacillus amyloliquefaciens, Bacillus jelly-like and Streptomyces lividans to obtain biological organic Fat.