CN111620745B - Method for producing bio-organic fertilizer by degrading agricultural wastes with biological agent - Google Patents
Method for producing bio-organic fertilizer by degrading agricultural wastes with biological agent Download PDFInfo
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- CN111620745B CN111620745B CN202010387541.XA CN202010387541A CN111620745B CN 111620745 B CN111620745 B CN 111620745B CN 202010387541 A CN202010387541 A CN 202010387541A CN 111620745 B CN111620745 B CN 111620745B
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C11/00—Other nitrogenous fertilisers
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Plant Pathology (AREA)
- Fertilizers (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent. The biological organic fertilizer is obtained by mixing fresh cow dung, lignocellulose waste, dried vinasse and L-alanine fermentation waste liquid, adjusting the carbon-nitrogen ratio to 25-30%, the water content to 55-65%, adding 0.1-0.5% of a biological microbial inoculum, and then carrying out decomposition fermentation for 6-18 days. The invention provides nutrient substances and growth promoting factors required in the functional microbial fermentation process by using the dried distiller's grains and the enzymatic hydrolysate thereof, improves the number of effective viable bacteria in the biological microbial inoculum, and reduces the nutrition difference of the decomposed fermentation environment; the mycoprotein and the L-alanine components in the L-alanine fermentation waste liquid are utilized to accelerate the spore germination. The two effectively supplement various amino acids, vitamins and calcium-phosphorus mineral components, improve the nutritional quality and functional potency of the bio-organic fertilizer, and are beneficial to increasing the yield and income of crops and improving the quality.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent.
Background
With the rapid development of modern agriculture in China, a large amount of agricultural wastes such as straws, feces of cultured livestock and poultry and the like are generated, the corresponding environmental pollution problem is gradually aggravated, and the sustainable development of society and economy is not facilitated. Wherein, the annual output of agricultural straw lignocellulose waste in China exceeds 9 hundred million tons, the annual output of livestock and poultry manure reaches 38 hundred million tons, and the resource utilization potential is huge. The lignocellulose waste contains rich organic matters, cellulose, crude protein, crude fat, nitrogen, phosphorus, potassium, calcium, magnesium, sulfur and other nutrient components, and 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 manure also contains various nutrient elements such as nitrogen, phosphorus, potassium and the like which are necessary for crops, and the nutrient content of 1 ton of manure is equivalent to 20-30 kg of fertilizer. According to the circular economy concept of 'reduction, reutilization and resource' in China, the agricultural waste resources are utilized to produce the green biological organic fertilizer, the popularization and the use of commercial organic fertilizers are supported, the green sustainable development targets of 'waste recycling, resource efficient utilization and clean and controllable production' in modern agriculture are effectively promoted, and the circular economy development of the modern agriculture is promoted.
The waste biomass is decomposed and fermented into fertilizer, organic materials such as livestock manure and the like are promoted to be decomposed and converted into fertilizer nutrients required by crops through the catabolism of microorganisms, pathogenic microorganisms such as bacteria and fungi and the like, spores and ova of the pathogenic microorganisms are killed, and the addition of a compound microbial inoculum containing multiple functional microorganisms can obviously improve the decomposition process and the quality of the organic fertilizer. At present, functional microorganisms widely used in the decomposing fermentation are mainly derived from bacillus, trichoderma, aspergillus and saccharomyces. The microorganisms of bacillus, trichoderma and aspergillus are added in the form of spores, in the actual decomposing fermentation environment, macromolecular organic substances need to be degraded into micromolecular substances, and nutrient substances such as carbohydrate carbon sources, amino acids, growth promoting factors and the like are insufficient, so that the problems of low organic matter degradation efficiency, insufficient decomposition, long fermentation time, incomplete organic fertilizer nutrients and the like caused by the insufficient germination, growth and proliferation of the spores are not facilitated, and the problems need to be solved urgently.
Taking cow dung decomposition fermentation as an example, most of the current research reports focus on optimizing the aspects of auxiliary material selection, material granularity, stack water content, carbon-nitrogen ratio, functional microorganism screening, microbial inoculum preparation and the like. Zhang Yufeng, etc[1]The optimal selection of corn straws, furfural residues and mushroom residues as auxiliary materials for the decomposing fermentation of the cow dung is explored by utilizing an aerobic fermentation mode after the addition of the biological agent, and the finding that the combination of the cow dung, the furfural residues and the straws is fast in temperature rise and long in high-temperature maintaining time, the total nutrient mass fraction of the decomposed organic fertilizer reaches the highest position of 5.04% under the combined condition of the cow dung, the straws and the urea, and the decomposing fermentation time is 23 days. Zhang He et al[2]The research and report of the influence of different carbon-nitrogen ratios on the aerobic decomposition process of cow dung show that in the actual production, the carbon-nitrogen ratio of the cow dung to the straw mixed material is 25-30: 1 is beneficial to the compost maturity and nutrient maintenance. Han Xianglong et al[3]The influence of different carbon-nitrogen ratios on the tobacco stem and cow dung decomposition process is also researched and reported, and the result shows that when the carbon-nitrogen ratio of the stacked material is 22: when 1 hour, the fertilizer has the highest content of nutrient elements such as total nitrogen, total phosphorus, total potassium and the like, and the comprehensive effect of decomposition and fermentation is the best. Zhang Yufeng, etc[4]The screening of the aerobic fermentation microbial inoculum for mixing and decomposing the cow dung and the corn straws is researched, and experimental results show that the cow dung and the corn straws have obvious decomposition effect and about 23 days of decomposition and fermentation time by using the composite microbial inoculum consisting of aspergillus and saccharomycetes for efficiently degrading cellulose and lignin. Tianzhihui, etc[5]The research on the initial conditions of the aerobic decomposition and fermentation of the cow dung and the straws shows that the water content of the stack is adjusted to be about 60 percent, and the carbon-nitrogen ratio is 30: about 1, the size of the crushed wheat straw is less than or equal to 1cm, and the effect is optimal. Patent CN110668861A discloses a method for producing an organic fertilizer by decomposing and fermenting myriophyllum viridissimum and cow dung, wherein the raw materials comprise 30-70% of fresh myriophyllum viridissimum, 12-52% of cow dung, 5% of straws, 10% of sawdust and 3% of waste biomass carbon, the water content is adjusted to be 60-70%, and the carbon-nitrogen ratio is adjusted to be 25-30: the composite microbial inoculum consists of a bacillus subtilis, bacillus licheniformis, saccharomyces cerevisiae and trichoderma viride spore suspension, the decomposing fermentation process is divided into two stages, firstly aerobic fermentation is carried out for 9-12 days, then secondary fermentation is carried out for 21-27 days, and the decomposing fermentation time is slightly long.
The above statements in the background are merely intended to facilitate a thorough understanding of the present disclosure (including the technical means used, technical problems solved and technical advantages brought about) and should not be taken as an acknowledgement or any form of suggestion that this information forms part of the prior art already known to a person skilled in the art.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a method for producing a biological organic fertilizer by degrading agricultural wastes by using a biological microbial inoculum, and aims to solve the problems of slow germination, growth and proliferation of microbial spores, low raw material degradation efficiency, insufficient decomposition, long fermentation time, incomplete organic fertilizer nutrients and the like caused by the defects of nutrient substances and growth promoting factors in a decomposition fermentation environment in the prior decomposition fermentation technology. Meanwhile, a new way is provided for the full utilization of mycoprotein and residual L-alanine in the L-alanine fermentation waste liquid, and the treatment cost of the L-alanine fermentation waste liquid is reduced. The production efficiency, the nutritional quality and the functional potency of the bio-organic fertilizer are improved by using the distiller's dried grains and the L-alanine fermentation waste liquid.
In order to solve the technical problems, the invention adopts the following technical scheme:
the invention provides a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent, which comprises the following steps:
s1: uniformly mixing fresh cow dung, lignocellulose waste and dried vinasse to form a pile, and adjusting the carbon-nitrogen ratio of the pile to 25-30;
s2: adding the L-alanine fermentation waste liquid into the pile obtained in the step S1, uniformly stirring, and controlling the pH7-8 of the pile added with the L-alanine fermentation waste liquid and the water content of 55-65%;
s3: adding a biological agent into the pile added with the L-alanine fermentation waste liquid obtained in the step S2, and uniformly stirring, wherein the usage amount of the biological agent is 0.1-0.5% of the total mass of the pile added with the L-alanine fermentation waste liquid;
s4: fermenting for 15-18 days by adopting 100kg scale static state maturity, or fermenting for 6-8 days by adopting 1-10 ton scale bioreactor, and then adding biocontrol probiotics to obtain the biological organic fertilizer, wherein the effective viable count in the fertilizer is not less than 0.2 hundred million/g.
In the above technical scheme, further, the lignocellulosic waste comprises one or more of corn stover, wheat straw, rice straw, sorghum stover, rice straw, wheat bran, rice husk, corncob and rice bran, and is pulverized to 1-2cm before mixing.
In the technical scheme, the biocontrol probiotics are selected from one or more of bacillus subtilis, bacillus firmus, paenibacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus.
In the technical scheme, the L-alanine fermentation waste liquid is liquid which is generated after L-alanine is extracted through fermentation of escherichia coli, escherichia coli or pseudomonas and contains mycoprotein and residual L-alanine.
In the technical scheme, furthermore, the mass of the dried distillers' grains is 5% -10% of that of the fresh cow dung.
In the technical scheme, the biological agent is prepared by directly mixing bacillus subtilis fermentation liquor, aspergillus niger fermentation liquor, trichoderma reesei fermentation liquor and saccharomyces cerevisiae fermentation liquor, and the effective viable count in the biological agent is more than or equal to 6.6 multiplied by 109CFU/mL. The effective viable count in the bacillus subtilis fermentation liquor is more than or equal to 1.6 multiplied by 1010CFU/mL, the number of effective viable bacteria in Aspergillus niger fermentation liquor is more than or equal to 3.2 multiplied by 109CFU/mL, the effective viable count in the Trichoderma reesei fermentation broth is more than or equal to 3.6 multiplied by 109CFU/mL, effective viable count in Saccharomyces cerevisiae fermentation broth is not less than 4.8 × 109CFU/mL。
In the above technical solution, further, the culture medium for preparing the fermentation broth of bacillus subtilis, aspergillus niger, trichoderma reesei and saccharomyces cerevisiae comprises:
(1) the bacillus subtilis culture medium: 30-70g/L of glucose, 5-15g/L of peptone, 10-30 g/L of beef extract, 5-15g/L of yeast powder, 5-15g/L of calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate, wherein the pH is 7-8;
(2) aspergillus niger and Trichoderma reesei media: 30-60g/L of glucose, 5-15g/L of malt extract powder, (NH)4)2SO45-15g/L of calcium carbonate, 5-15g/L of calcium carbonate and the balance of dried distillers' grains enzymatic hydrolysate, wherein the pH is 7-8;
(3) a saccharomyces cerevisiae culture medium: 60-120g/L of glucose, 5-15g/L of yeast powder, 20-40 g/L of peptone, 5-15g/L of calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate with the pH value of 6-7.
The dried distiller's grains enzymatic hydrolysate is prepared by the following method: the method comprises the steps of uniformly mixing 5-10% of dried vinasse by mass with water, adjusting the pH value to 8-11 by using an ammonia water solution, carrying out water bath at 80 ℃ for 60 minutes, adding alkaline protease into the dried vinasse at a ratio of 200-600U/g at 50 ℃ for enzymolysis for 24 hours, centrifuging, removing precipitates, and obtaining supernatant which is dried vinasse enzymolysis liquid.
In the above technical solution, further, during the static decomposing fermentation process in the 100kg scale, the stack is not turned before the temperature is raised to 55 ℃, and after the temperature is raised to 65 ℃, the stack is turned every day to keep the temperature not higher than 70 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days. Decomposing and fermenting for 15-18 days to obtain mature organic fertilizer, and adding biocontrol probiotics such as one or more of bacillus subtilis, bacillus firmus, paenibacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus to obtain the biological 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 ℃, and the control is removed after the temperature is raised; the ventilation frequency is 2-3 times per day; stirring for 2-3 times every day at the rotating speed of 50-150 rpm; after fermenting for 6-8 days, dewatering or air drying by a machine to obtain a mature organic fertilizer; and adding biocontrol probiotics such as one or more of bacillus subtilis, bacillus firmus, paenibacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus to obtain the biological organic fertilizer.
The bio-organic fertilizer produced by the technical scheme has slight fermentation faint scent, and all indexes reach or exceed the national industrial standard (NY 525-.
Compared with the prior art, the invention has the beneficial effects that: the invention uses the distiller's dried grain to provide nutrient substances and growth promoting factors required by fermentation and decomposed fermentation for functional microorganisms, and reduces the nutrition difference in the growth microenvironment; on the other hand, mycoprotein and L-alanine components in the L-alanine fermentation waste liquid are effectively utilized to accelerate spore germination of spore-producing microorganisms, and the fermentation waste liquid is used for replacing other water sources, so that water is saved; on the basis, the bioreactor has the advantages of thorough decomposition, rapid propagation of functional microorganisms, better control of fermentation conditions (such as ventilation, stirring, nutrition transmission and the like) and the like, the decomposition fermentation time is greatly reduced from 30-40 days to 6-18 days, and the dried distillers' grains and the L-alanine fermentation waste liquid also effectively supplement more than ten kinds of amino acids (such as alanine, arginine, aspartic acid, cysteine, glutamic acid, leucine and the like), vitamins and calcium-phosphorus mineral compositions, improve the organic matter content and the total nutrient content of the organic fertilizer, improve the nutrient quality and the functional potency, and are beneficial to increasing the yield and the income of crops.
Detailed Description
The present invention is further described in the following examples, it is to be understood that the scope of the claims of the present invention is not limited to the specific embodiments described, which are provided as examples of the present invention, and the skilled person can easily modify the specific embodiments of the present invention or make equivalent substitutions for some technical features with reference to the description of the present specification, and such modifications and substitutions without inventive effort are also within the scope of the claims of the present invention. Materials of the same or similar type, quality, nature or function as the following reagents and instruments may be used in the practice of the present invention. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The strains, reagents and raw materials used in the following examples of the invention were derived as follows:
the used bacillus subtilis ACCC 19742, aspergillus niger ACCC 32589 and saccharomyces cerevisiae ACCC 21162 are all purchased from China agricultural microbial strain preservation management center, and the used trichoderma reesei Rut C30(NRRL 11460) is given to American agricultural research strain preservation center.
The biochemical reagents such as glucose, peptone, beef extract, yeast powder, alkaline protease and the like are purchased from bio-engineering (Shanghai) GmbH, and the raw materials such as fresh cow dung, dried distiller's grains, straws, wheat bran, rice hulls, L-alanine fermentation waste liquid and the like are provided by Dalian gold Weide Biotech Co.
Example 1:
a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent comprises the following specific steps:
1. preparing dried vinasse enzymatic hydrolysate:
mixing the distiller's dried grains with tap water in a mass ratio of 10%, adjusting the pH to 9 with ammonia water solution, carrying out water bath at 80 ℃ for 60 minutes, adding alkaline protease into the distiller's dried grains at a ratio of 400U/g for enzymolysis at 50 ℃ for 24 hours, and centrifuging to obtain the distiller's dried grains enzymatic hydrolysate.
2. Strain activation
Coating the bacillus subtilis on an LB (Luria-Bertani Agar) culture medium, and culturing for 20 hours at 30 ℃ to obtain a bacillus subtilis colony; respectively coating Aspergillus niger and Trichoderma reesei on a PDA (Potato Dextrose agar) culture medium, and culturing at 30 ℃ for 96 hours to obtain Aspergillus niger and Trichoderma reesei spores; saccharomyces cerevisiae was spread on YPD (Yeast Extract Peptone Dextrose agar) medium and cultured at 30 ℃ for 20 hours to obtain a Saccharomyces cerevisiae colony.
3. Fermentation culture
(1) Selecting activated Bacillus subtilis single colony, inoculating in liquid fermentation culture medium, culturing at constant temperature of 150rpm and 30 deg.C for 24 hr to obtain Bacillus subtilis fermentation broth with effective viable count of 1.6 × 1010CFU/mL; wherein, the fermentation medium comprises the following components: 60g/L of glucose, 15g/L of peptone, 30g/L of beef extract, 15g/L of yeast powder, 15g/L of calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate, and the pH value is 7.2.
(2) Collecting activated Aspergillus niger spore with sterile water, inoculating into liquid fermentation culture medium, culturing at constant temperature of 30 deg.C at 150rpm for 72 hr to obtain Aspergillus niger fermentation liquid with effective viable count of 3.2 × 109CFU/mL; wherein, the fermentation medium comprises the following components: 50g/L glucose, 15g/L malt extract powder, (NH)4)2SO415g/L, 15g/L calcium carbonate, and the balance of dried distillers' grains enzymatic hydrolysate, and the pH value is 7.2.
(3) Collecting the activated trichoderma reesei spores with sterile water, inoculating the collected trichoderma reesei spores into a liquid fermentation medium, culturing at constant temperature of 30 ℃ for 72 hours at 150rpm to obtain trichoderma reesei fermentation liquor, wherein the effective viable count is 3.6 multiplied by 109CFU/mL; wherein,the fermentation medium comprises the following components: 50g/L glucose, 15g/L malt extract powder, (NH)4)2SO415g/L, 15g/L calcium carbonate, and the balance of dried distillers' grains enzymatic hydrolysate, and the pH value is 7.2.
(4) Selecting activated Saccharomyces cerevisiae single colony, inoculating in liquid fermentation culture medium, culturing at constant temperature of 150rpm and 30 deg.C for 24 hr to obtain Saccharomyces cerevisiae fermentation liquid with effective viable count of 4.8 × 109CFU/mL; wherein, the fermentation medium comprises the following components: 100g/L glucose, 15g/L yeast powder, 40g/L peptone, 15g/L calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate, and the pH value is 6.5.
4. Mixing the fermentation liquors
Uniformly mixing the bacillus subtilis fermentation broth, the aspergillus niger fermentation broth, the trichoderma reesei fermentation broth and the saccharomyces cerevisiae fermentation broth to obtain the biological agent, wherein the effective viable count in the biological agent is 6.6 multiplied by 109CFU/mL。
Control experiment:
1. strain activation
Coating the bacillus subtilis on an LB (Luria-Bertani Agar) culture medium, and culturing for 20 hours at 30 ℃ to obtain a bacillus subtilis colony; respectively coating Aspergillus niger and Trichoderma reesei on a PDA (Potato Dextrose agar) culture medium, and culturing at 30 ℃ for 96 hours to obtain Aspergillus niger and Trichoderma reesei spores; saccharomyces cerevisiae was spread on YPD (Yeast Extract Peptone Dextrose agar) medium and cultured at 30 ℃ for 20 hours to obtain a Saccharomyces cerevisiae colony.
2. Fermentation culture
(1) Selecting activated Bacillus subtilis single colony, inoculating in liquid fermentation culture medium, culturing at constant temperature of 150rpm and 30 deg.C for 24 hr to obtain Bacillus subtilis fermentation broth with effective viable count of 7.5 × 109CFU/mL; wherein, the fermentation medium comprises the following components: 60g/L of glucose, 15g/L of peptone, 30g/L of beef extract, 15g/L of yeast powder, 15g/L of calcium carbonate and the balance of tap water, wherein the pH value is 7.2.
(2) Collecting activated Aspergillus niger spore with sterile water, inoculating into liquid fermentation culture medium, culturing at constant temperature of 30 deg.C at 150rpm for 72 hr to obtain Aspergillus niger fermentation liquid with effective viable count of 4.2 × 108CFU/mL; wherein, the fermentation culture is carried outThe nutrient medium comprises the following components: 50g/L glucose, 15g/L malt extract powder, (NH)4)2SO415g/L, 15g/L calcium carbonate, and the balance of tap water, and the pH value is 7.2.
(3) Collecting the activated trichoderma reesei spores with sterile water, inoculating the collected trichoderma reesei spores into a liquid fermentation medium, culturing at constant temperature of 30 ℃ for 72 hours at 150rpm to obtain trichoderma reesei fermentation liquor, wherein the effective viable count is 4.5 multiplied by 108CFU/mL; wherein, the fermentation medium comprises the following components: 50g/L glucose, 15g/L malt extract powder, (NH)4)2SO415g/L, 15g/L calcium carbonate, and the balance of tap water, and the pH value is 7.2.
(4) Selecting activated Saccharomyces cerevisiae single colony, inoculating in liquid fermentation culture medium, culturing at constant temperature of 150rpm and 30 deg.C for 24 hr to obtain Saccharomyces cerevisiae fermentation liquid with effective viable count of 1.0 × 109CFU/mL; wherein, the fermentation medium comprises the following components: 100g/L glucose, 15g/L yeast powder, 40g/L peptone, 15g/L calcium carbonate and the balance tap water, and the pH value is 6.5.
3. Mixing the fermentation liquors
Uniformly mixing the bacillus subtilis fermentation broth, the aspergillus niger fermentation broth, the trichoderma reesei fermentation broth and the saccharomyces cerevisiae fermentation broth to obtain the biological agent, wherein the effective viable count is 2.2 multiplied by 109CFU/mL。
According to the invention, rich nutrients and growth promoting factors in the dried distiller's grain enzymatic hydrolysate are utilized to perform functional microbial fermentation culture, compared with bacillus subtilis, aspergillus niger, trichoderma viride and saccharomyces cerevisiae fermentation liquor prepared by a conventional method, the effective viable count is greatly increased, and the effective viable count in the prepared biological agent is increased by 3 times.
Example 2:
a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent comprises the following specific decomposition and fermentation steps:
1. mixing and piling
Mixing fresh cow dung, crushed straws and dried vinasse, and adjusting the carbon-nitrogen ratio to be 30, wherein the using amount of the dried vinasse is 10% of the mass of the fresh cow dung; adding L-alanine fermentation waste liquid, stirring uniformly, controlling the pH value to be 7.8, and controlling the water content to be 65%; then adjusting the water contentThe biological agent prepared in example 1 (effective viable count is 6.6 × 10) was added to 0.5% of the total mass of the rear stack9CFU/mL) and uniformly mixing, wherein the total mass of the pile is close to 100 kg.
2. Fermenting by decomposition
Static state decomposition fermentation is carried out on a 100kg scale, and when the temperature is raised to 55 ℃, the stack is not turned; turning over the pile every day to keep the temperature not higher than 70 ℃ after the temperature is increased to 65 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days; when the temperature is reduced to 35 ℃, turning is stopped, and the compost is fermented for 15 days to obtain a mature organic fertilizer, and then a biocontrol probiotic bacillus firmus agent is added to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.5 hundred million/g, the mass fraction of organic matters is 62.5%, the mass fraction of total nutrients is 9.2%, the mass fraction of water is 27.6%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 2/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 7.8, and all indexes reach or exceed the national industrial standard (NY 525-. After the biological organic fertilizer is applied, pests, ova and larvae such as root-knot nematodes, cyst nematodes, golden nematodes, heterodera and the like parasitic on roots of crops can be quickly inhibited and killed, and meanwhile, rich organic matters, nitrogen, phosphorus, potassium, amino acids, vitamins, mineral substances and other nutrients are provided for the crops.
Control experiment:
1. mixing and piling
Mixing fresh cow dung and crushed straws, adding tap water, adjusting the carbon-nitrogen ratio to be 30, adjusting the pH to be 7.8, and controlling the water content to be 65%; the biological agent (effective viable count of 2.2X 10) prepared in example 1 was added to the pile in an amount of 0.5% of the total mass after adjusting the water content9CFU/mL) and uniformly mixing, wherein the total mass of the pile is close to 100 kg.
2. Fermenting by decomposition
Static state decomposition fermentation is carried out on a 100kg scale, and when the temperature is raised to 55 ℃, the stack is not turned; turning over the pile every day to keep the temperature not higher than 70 ℃ after the temperature is increased to 65 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days; when the temperature is reduced to 35 ℃, turning is stopped, and the compost is fermented for 30 days to obtain a mature organic fertilizer, and then a biocontrol probiotic bacillus firmus agent is added to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.5 hundred million/g, the mass fraction of organic matters is 50.0%, the mass fraction of total nutrients is 6.6%, the mass fraction of water is 28.7%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 5/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 8.0, and all indexes also reach or exceed the national organic fertilizer industry standard (NY 525. supplement 2012) and the agricultural industry bio-organic fertilizer standard (NY 884. supplement 2012) of the people's republic of China. However, in the embodiment 2, the time for decomposition and fermentation is only 15 days, the production efficiency of the biological organic fertilizer is improved by 1 time, and the nutritional quality and the functional potency of the biological organic fertilizer are also greatly improved.
Example 3:
a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent comprises the following specific decomposition and fermentation steps:
1. mixing and piling
Mixing fresh cow dung, crushed wheat bran and dried distiller grains, and adjusting the carbon-nitrogen ratio to 25, wherein the consumption of the dried distiller grains is 5% of the mass of the fresh cow dung; adding L-alanine fermentation waste liquid, stirring uniformly, controlling the pH value to 7.8, and controlling the water content to 55%; and then adding the biological agent (the effective viable count is 6.6 multiplied by 109CFU/mL) prepared in the example 1 into the pile body according to 0.1 percent of the total mass of the pile body after the water content is adjusted, and uniformly mixing, wherein the total mass of the pile body is close to 100 kg.
2. Fermenting by decomposition
Static state decomposition fermentation is carried out on a 100kg scale, and when the temperature is raised to 55 ℃, the stack is not turned; turning over the pile every day to keep the temperature not higher than 70 ℃ after the temperature is increased to 65 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days; when the temperature is reduced to 35 ℃, turning is stopped, and the compost is fermented for 18 days to obtain a mature organic fertilizer, and then the biocontrol probiotic streptomyces microflavus is added to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.3 hundred million/g, the mass fraction of organic matters is 60.4%, the mass fraction of total nutrients is 8.4%, the mass fraction of water is 25.9%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 2/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 7.5, and all indexes reach or exceed the national industrial standard (NY 525-. After the biological organic fertilizer is applied, the soil fertility can be improved, the propagation of pathogenic microorganisms is inhibited, the rooting, the germination and the maturation of crops are promoted, the yield and the income are increased, the quality of agricultural products is improved and the like, and meanwhile, rich nutrients such as organic matters, nitrogen, phosphorus, potassium, amino acids, vitamins, mineral substances and the like are provided for the crops.
Control experiment:
1. mixing and piling
Mixing fresh cow dung and crushed wheat bran, adding tap water, adjusting the carbon-nitrogen ratio to be 25, adjusting the pH to be 7.8, and controlling the water content to be 55%; the biological agent (the effective viable count is 2.2 multiplied by 109CFU/mL) prepared in the example 1 is added according to 0.1 percent of the total mass of the pile after the water content is adjusted, and the mixture is uniformly mixed, wherein the total mass of the pile is close to 100 kg.
2. Fermenting by decomposition
Static state decomposition fermentation is carried out on a 100kg scale, and when the temperature is raised to 55 ℃, the stack is not turned; turning over the pile every day to keep the temperature not higher than 70 ℃ after the temperature is increased to 65 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days; when the temperature is reduced to 35 ℃, turning is stopped, and the compost is fermented for 35 days to obtain a mature organic fertilizer, and then a biocontrol probiotic bacillus firmus agent is added to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.2 hundred million/g, the mass fraction of organic matters is 48.8%, the mass fraction of total nutrients is 6.1%, the mass fraction of water is 23.7%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 5/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 7.6, and all indexes also reach or exceed the national organic fertilizer industry standard (NY 525. supplement 2012) and the agricultural industry bio-organic fertilizer standard (NY 884. supplement 2012) of the people's republic of China. However, in the embodiment 3, the decomposing fermentation time is only 18 days, the production efficiency of the biological organic fertilizer is improved by nearly 1 time, and the nutritional quality and the functional potency are also greatly improved.
Example 4:
a method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent comprises the following specific decomposition and fermentation steps:
1. mixing and piling
Mixing fresh cow dung, crushed rice hulls and dried vinasse, and adjusting the carbon-nitrogen ratio to be 28, wherein the using amount of the dried vinasse is 8% of the mass of the fresh cow dung; adding L-alanine fermentation waste liquid, stirring uniformly, controlling the pH value to 7.8, and controlling the water content to 60%; then the biological agent (the effective viable count is 6.6 multiplied by 10) prepared in the example 1 is added according to 0.3 percent of the total mass of the pile after the water content is adjusted9CFU/mL), mixed uniformly, the total mass is close to 100 kg.
2. Fermenting by decomposition
Fermenting by adopting a 5-ton scale bioreactor, controlling the initial fermentation temperature to be 30 ℃, and removing control after heating; the ventilation frequency is 3 times/day; stirring for 3 times every day at the rotating speed of 100 rpm; controlling the temperature not to exceed 70 ℃ in the whole process, and removing water or air-drying by a machine after the temperature in the stack body is reduced to 35 ℃ for 7 days to obtain a mature organic fertilizer; and adding biocontrol probiotic bacillus polymyxa to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.5 hundred million/g, the mass fraction of organic matters is 58.6%, the mass fraction of total nutrients is 8.8%, the mass fraction of water is 29.3%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 2/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 7.9, and all indexes also reach or exceed the national organic fertilizer industry standard (NY 525. supplement 2012) and the agricultural industry bio-organic fertilizer standard (NY 884. supplement 2012) of the people's republic of China. After the biological organic fertilizer is applied, bacterial and fungal soil-borne diseases of plants, such as bacterial wilt and fusarium wilt of the plants, can be effectively prevented and controlled, so that bacterial and fungal diseases of plant leaves are obviously reduced, and the biological organic fertilizer has obvious growth promoting and yield increasing effects on crops. Meanwhile, the fertilizer provides rich nutrients such as organic matters, nitrogen, phosphorus and potassium, amino acids, vitamins, mineral substances and the like for crops.
Control experiment:
1. mixing and piling
Mixing fresh cow dung and crushed rice hulls, adding tap water, adjusting the carbon nitrogen ratio to be 28, adjusting the pH value to be 7.8, and controlling the water content to be 60%; the biological agent (effective viable count of 2.2X 10) prepared in example 1 was added to the pile in an amount of 0.3% of the total mass after adjusting the water content9CFU/mL) and uniformly mixing, wherein the total mass of the pile is close to 100 kg.
2. Fermenting by decomposition
Fermenting by adopting a 5-ton scale bioreactor, controlling the initial temperature to be 30 ℃, and removing the control after the temperature is raised; the ventilation frequency is 3 times/day; stirring for 3 times every day at the rotating speed of 100 rpm; the temperature is controlled not to exceed 70 ℃ in the whole process, when the temperature in the stack body is reduced to 35 ℃, the machine is dewatered or air-dried, and mature organic fertilizer can be obtained in 12 days; and adding biocontrol probiotic bacillus polymyxa to obtain the biological organic fertilizer. The number of effective viable bacteria in the produced bio-organic fertilizer is more than 0.5 hundred million/g, the mass fraction of organic matters is 50.1%, the mass fraction of total nutrients is 6.5%, the mass fraction of water is 29.5%, the death rate of roundworm eggs reaches 100%, the number of faecal coliform groups is less than 2/g, no total arsenic, mercury, lead, cadmium and chromium are detected, the pH value is 7.7, and all indexes also reach or exceed the national organic fertilizer industry standard (NY 525. supplement 2012) and the agricultural industry bio-organic fertilizer standard (NY 884. supplement 2012) of the people's republic of China. However, in the embodiment 4, the decomposing fermentation time is only 7 days, the production efficiency of the biological organic fertilizer is improved by more than 70%, and the nutritional quality and the functional potency are also greatly improved.
It will be apparent to those skilled in the art from this disclosure that many changes and modifications can be made, or equivalents modified, in the embodiments of the invention without departing from the scope of the invention. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention shall still fall within the protection scope of the technical solution of the present invention, unless the contents of the technical solution of the present invention are departed.
Reference to the literature
1. Zhangyufeng, cautious field, frontier, great forest, great waves, Dongliang, Liumegahui, Guhong sea, Gangqingju, the influence of organic materials on the aerobic composting process of cow dung, Shandong agricultural science, 2019,51(05):76-82.
2. The influence of different carbon and nitrogen ratios on the aerobic composting process of cow dung is reported in university of Gansu, 2019,54(01):60-67.
3. Korea dragon, Wuwei Wei, Zhao Peng Bo, Wei Cheng, Yuan Shuai, Li Si Chun, Mao Karan, ox Huali, Zhang Lixin the influence of different carbon-nitrogen ratios on the tobacco stem and cow dung composting process Jiangsu agricultural science 2019,47(16): 303-.
4. Zhangyufeng, cautious field, frontier, Guohuanhai, far-reaching palace, Liumegahui, Lirui, Chenjiaqiu, Lujia method, screening aerobic fermentation bacteria agent of mixed compost of cow dung and corn straw, Chinese soil and fertilizer, 2019, 172-class 178.
5. Tianzhihui, Wanyani, Zhonghuachao, Lujundang, research on initial conditions of aerobic fermentation composting of cow dung and straw, China soil and fertilizer, 2018(06):192 + 197.
Claims (5)
1. A method for producing a biological organic fertilizer by degrading agricultural wastes with a biological agent is characterized by comprising the following steps:
s1: uniformly mixing fresh cow dung, lignocellulose waste and dried vinasse to form a pile, and adjusting the carbon-nitrogen ratio of the pile to 25-30;
s2: adding the L-alanine fermentation waste liquid into the pile obtained in the step S1, uniformly stirring, and controlling the pH6-8 of the pile added with the L-alanine fermentation waste liquid and the water content of 55-65%;
s3: adding a biological agent into the pile added with the L-alanine fermentation waste liquid obtained in the step S2, and uniformly stirring, wherein the usage amount of the biological agent is 0.1-0.5% of the total mass of the pile added with the L-alanine fermentation waste liquid;
s4: fermenting for 15-18 days by adopting 100kg scale static state maturity, or fermenting for 6-8 days by adopting 1-10 ton scale bioreactor, and then supplementing biocontrol probiotics to obtain the biological organic fertilizer, wherein the effective viable count in the fertilizer is not less than 0.2 hundred million/g;
the L-alanine fermentation waste liquid is liquid which is generated after the L-alanine is extracted through the fermentation of escherichia coli, escherichia coli or pseudomonas and contains mycoprotein and residual L-alanine;
the mass of the dried vinasse is 5-10% of that of the fresh cow dung;
the biological agent is prepared by directly mixing bacillus subtilis fermentation liquor, aspergillus niger fermentation liquor, trichoderma reesei fermentation liquor and saccharomyces cerevisiae fermentation liquor, and the effective viable count in the biological agent is more than or equal to 6.6 multiplied by 109CFU/mL; wherein the effective viable count in the bacillus subtilis fermentation broth is more than or equal to 1.6 multiplied by 1010CFU/mL, the number of effective viable bacteria in Aspergillus niger fermentation liquor is more than or equal to 3.2 multiplied by 109CFU/mL, the effective viable count in the Trichoderma reesei fermentation broth is more than or equal to 3.6 multiplied by 109CFU/mL, effective viable count in Saccharomyces cerevisiae fermentation broth is not less than 4.8 × 109 CFU/mL；
The preparation of the bacillus subtilis fermentation broth, the aspergillus niger fermentation broth, the trichoderma reesei fermentation broth and the saccharomyces cerevisiae fermentation broth is sequentially subjected to activation culture and fermentation culture, and the fermentation culture medium of each strain comprises:
(1) b, bacillus subtilis: 30-70g/L of glucose, 5-15g/L of peptone, 10-30 g/L of beef extract, 5-15g/L of yeast powder, 5-15g/L of calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate, wherein the pH is 7-8;
(2) aspergillus niger and trichoderma reesei: 30-60g/L of glucose, 5-15g/L of malt extract powder, (NH)4)2SO45-15g/L of calcium carbonate, 5-15g/L of calcium carbonate and the balance of dried distillers' grains enzymatic hydrolysate, wherein the pH is 7-8;
(3) and (3) saccharomyces cerevisiae: 60-120g/L of glucose, 5-15g/L of yeast powder, 20-40 g/L of peptone, 5-15g/L of calcium carbonate and the balance of dried distiller's grains enzymatic hydrolysate, wherein the pH is 6-7;
the distiller's dried grain enzymolysis liquid is prepared by the following method: the method comprises the steps of uniformly mixing 5-10% of dried vinasse by mass with water, adjusting the pH value to 8-11 by using an ammonia water solution, carrying out water bath at 80 ℃ for 60 minutes, adding alkaline protease into the dried vinasse at a ratio of 200-600U/g at 50 ℃ for enzymolysis for 24 hours, centrifuging, removing precipitates, and obtaining supernatant which is dried vinasse enzymolysis liquid.
2. The method for producing the bio-organic fertilizer by using the biological inoculant to degrade agricultural wastes, wherein the lignocellulose wastes comprise one or more of corn stalks, wheat stalks, rice stalks, sorghum stalks, rice straw stalks, wheat bran, rice husks, corncobs and rice bran, and the mixture is crushed to 1-2cm before being mixed.
3. The method for producing the bio-organic fertilizer by using the biological agent to degrade the agricultural wastes according to claim 1 or 2, wherein the biocontrol probiotic bacteria are selected from one or more of bacillus subtilis, bacillus firmus, paenibacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus.
4. The method for producing the bio-organic fertilizer by degrading agricultural wastes through the biological agent as claimed in claim 1 or 2, wherein during the static decomposing fermentation process in the 100kg scale, the stack is not turned before the temperature is increased to 55 ℃, and after the temperature is increased to 65 ℃, the stack is turned every day to keep the temperature not higher than 70 ℃; when the temperature is reduced to 50 ℃, turning the pile once every two days; decomposing and fermenting for 15-18 days to obtain mature organic fertilizer, and adding one or more of bacillus subtilis, bacillus firmus, paenibacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus to obtain the biological organic fertilizer.
5. The method for producing the bio-organic fertilizer by degrading agricultural wastes with the biological agent 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 ℃, and the control is removed after the temperature is raised; the ventilation frequency is 2-3 times per day; stirring for 2-3 times every day at the rotating speed of 50-150 rpm; after decomposing and fermenting for 6-8 days, dewatering or air drying by a machine to obtain a mature organic fertilizer; and adding one or more of bacillus subtilis, bacillus firmus, bacillus polymyxa, bacillus licheniformis, bacillus megaterium, bacillus amyloliquefaciens, bacillus mucilaginosus and streptomyces microflavus to obtain the biological organic fertilizer.
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| CN113636876A (en) * | 2021-07-22 | 2021-11-12 | 湖北田申甲生物环保科技有限公司 | Method for preparing biological fertilizer from agricultural production residual waste |
| CN114736830B (en) * | 2022-05-10 | 2023-07-14 | 中南大学 | Kitchen waste treatment composite microbial inoculant, preparation method thereof and method for producing organic fertilizer by using kitchen waste treatment composite microbial inoculant |
| CN115466138A (en) * | 2022-08-19 | 2022-12-13 | 武汉和信益科技有限公司 | Resource utilization method of liquid organic waste |
| CN115504841A (en) * | 2022-11-02 | 2022-12-23 | 安徽省农业科学院棉花研究所 | Fermented pig manure organic fertilizer and production method thereof |
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