CN112661547B - Composite biological decomposition agent and preparation method thereof - Google Patents
Composite biological decomposition agent and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a composite biological decomposition agent, which consists of the following components in parts by weight: 2-10 parts of clostridium paphiopedilum thermophilum, 1-5 parts of lactobacillus thermophilus, 10-30 parts of composite bacteria, 2-5 parts of a composite enzyme preparation, 2-5 parts of composite actinomycetes, 5-10 parts of composite fungi, 40-70 parts of an organic carrier and 0.001 part of tea tree essential oil; simultaneously discloses a preparation method of the composite biological decomposition agent. The invention can efficiently, quickly and thoroughly decompose organic materials, has the characteristics of quick temperature rise and long high-temperature duration, can effectively kill pathogenic bacteria and ova in the materials, greatly shortens one-time decomposition time, reduces about one week, does not contain harmful bacteria in the decomposed materials, and has the advantages that the composite bacterial flora in the product is mostly bacillus, and the bacterial flora can generate spores and is high-temperature resistant, so that the bacterial flora can be greatly reserved after the materials are decomposed, and the effect of the bacterial flora on soil and crops is exerted.
Description
Technical Field
The invention relates to the technical field of treatment and recycling of organic wastes, in particular to a composite biological decomposition agent and a preparation method thereof.
Background
In recent years, with the excessive and unreasonable application of chemical fertilizers, the soil plough layer structure of China suffers from serious damage, soil is poor, the soil fertility is low, various crop diseases and insect pests and soil-borne diseases are frequent, and the crop yield and the quality are influenced to a great extent. From the perspective of soil fertilization, as the input of chemical fertilizers is continuously increased, the environmental pollution caused by the chemical fertilizers is increasingly serious, and meanwhile, the input of organic fertilizers in the traditional agriculture is gradually lost, so that the soil fertility is greatly reduced, the content of organic matters in the soil is reduced, the input amount of the organic fertilizers in China is in a trend of reducing year by year, but the content of organic fertilizer resources in China and nutrient substances such as nitrogen, phosphorus, potassium and the like contained in the organic fertilizer resources in China is in a trend of increasing year by year, and the application of the organic fertilizers in addition is an important content for increasing the agricultural input at present. Therefore, the country strongly advocates the investment of organic fertilizers and bio-organic fertilizers, and insists on the matching use of organic fertilizers and inorganic fertilizers, so that the agricultural yield increase can be directly promoted, the utilization rate of the fertilizer is improved, the contradiction between supply and demand of the fertilizer is relieved, the input-output ratio is reduced, the agricultural production cost is reduced, and the method is not only beneficial to national economy, but also more beneficial to agricultural sustainable development. However, while many enterprises respond to national policies, the used organic materials are not thoroughly and effectively decomposed, and the materials which are not thoroughly decomposed also contain many adverse factors, including pathogenic bacteria, weed seeds, worm eggs, antibiotics, heavy metals, toxic and harmful gases and the like, and nutrient elements cannot be effectively degraded and cannot be absorbed by crops.
Many microorganisms in nature have the capacity of oxidizing and decomposing organic matters, and the microorganisms are utilized to carry out biochemical degradation on the organic matters under the conditions of certain temperature, humidity and pH value to form a substance similar to humus soil, so that the microorganism can be completely used as a fertilizer and can improve the soil thermodynamically.
Disclosure of Invention
The first problem to be solved by the present invention is: the composite biological decomposition agent can decompose organic materials efficiently, quickly and thoroughly, can quickly degrade cellulose, macromolecular protein, lignin, heavy metals and antibiotics in sludge, excrement and straws, is quick in temperature rise and long in high-temperature duration, can effectively kill pathogenic bacteria and ova in the materials, and greatly shortens one-time decomposition time.
The second problem to be solved by the present invention is: the preparation method comprises the steps of preparing a clostridium thermocellum microbial inoculum, and mixing the prepared clostridium thermocellum microbial inoculum with other components to obtain a finished product of the composite biological decomposition agent.
In order to solve the technical problems, the technical scheme of the invention is as follows:
the composite biological decomposition agent consists of the following components in parts by weight: 2-10 parts of clostridium papulophilus, 1-5 parts of lactobacillus thermophilus, 10-30 parts of composite bacteria, 2-5 parts of a complex enzyme preparation, 2-5 parts of composite actinomycetes, 5-10 parts of composite fungi, 40-70 parts of an organic carrier and 0.001 part of tea tree essential oil.
Preferably, the composite biological decomposition agent consists of the following components in parts by weight: 6 parts of clostridium paphiolyticum, 3 parts of lactobacillus thermophilus, 20 parts of composite bacteria, 3.5 parts of a composite enzyme preparation, 3.5 parts of composite actinomycetes, 7.5 parts of composite fungi, 55 parts of an organic carrier and 0.001 part of tea tree essential oil.
Preferably, the compound bacteria comprise one or more of bacillus subtilis, bacillus amyloliquefaciens, pseudomonas and bacillus pumilus; the compound enzyme preparation comprises one or more of acid protease, endo-cellulase and xylanase; the compound actinomycetes comprises one or more of Mycobacterium, Micromonospora, Streptomyces melanioides and Streptomyces rochei; the composite fungus comprises one or more of Trichoderma viride, Trichoderma reesei and Aspergillus niger; the organic carrier comprises one or more of rice hull powder, bran and straw powder.
A preparation method of a composite biological decomposition agent specifically comprises the following steps:
a. preparation of clostridium paphiolyticum: (1) seed culture: inoculating the bacterial colony on the plate culture medium into a seed culture solution, and performing shake cultivation at 55-60 ℃ for 16-24 h at the rotating speed of 200-260 rpm;
wherein, the seed culture solution is: 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water, wherein the pH value is adjusted to be 6.6-7.0;
(2) and (3) amplification culture: inoculating the seed culture solution into the expanded culture solution in an inoculation amount of 1-5%, and performing shake cultivation at 55-60 ℃ for 16-24 h at a rotation speed of 200-260 rpm;
wherein the expanding culture solution is 0.01g of formaldehyde, 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water for supplementing 100g, and the pH value is adjusted to be 6.6-7.0;
(3) preparation of a microbial inoculum: adding 0.1-0.5% by weight of protective agent and 10-30% by weight of carrier adsorbent into the product cultured in the step (2), and fully drying to obtain a powdery microbial inoculum;
wherein the protective agent is one or more of skimmed milk, soybean protein and whey powder, and the carrier adsorbent is one or more of talcum powder, light calcium carbonate and beneficial powder;
b. and b, adding the clostridium thermophilum cyperus microbial inoculum prepared in the step a into a pre-prepared organic carrier according to a proportion, sequentially and respectively adding lactobacillus thermophilus, compound bacteria, a compound enzyme preparation, compound actinomycetes and compound fungi under the condition of continuous stirring, spraying 0.001 weight percent of tea tree essential oil solution in the process of adding all thalli and uniformly mixing, and fully stirring and adsorbing to obtain the microbial agent.
Preferably, the stirring speed in the step b is 100-200 rmp, and each group of bacteria is stirred for more than 5min during adding.
Due to the adoption of the technical scheme, the invention has the beneficial effects that:
the composite biological decomposition agent is a novel product developed aiming at the rapid and efficient decomposition of livestock and poultry manure, sludge, crop straws, household garbage and other materials. The main functional bacterium clostridium thermocellum of the product is a strain with the number of UBA305 from southern American biotechnology and applied microbiological resource center preservation, the strain has high temperature resistance and strong activity, can continuously grow and reproduce at high temperature, and greatly improves the capability of decomposing cellulose and killing parasites and pathogenic bacteria in compost by metabolic products after the directional induction culture of an enlarged culture medium containing formaldehyde.
The product of the invention has rich strains, forms protective films among bacterial colonies under the protection of the organic adsorption carrier, does not interfere with each other, can preserve the activity to a greater extent and plays respective functions. The tea tree essential oil in the product is a protective agent and a bactericide, can effectively isolate various strains, and each strain forms a protective film to block and kill harmful pathogenic bacteria, so that the activity among the strains is greatly preserved.
The composite biological decomposition agent can decompose organic materials efficiently, quickly and thoroughly, can quickly degrade cellulose, macromolecular protein, lignin, heavy metal and antibiotics in sludge, excrement and straws, is quick in temperature rise and long in high-temperature duration, can effectively kill pathogenic bacteria and ova in the materials, greatly shortens one-time decomposition time, reduces about one week, and ensures that the decomposed materials do not contain harmful bacteria.
Drawings
The invention is further illustrated with reference to the following figures and examples.
FIG. 1 is a temperature profile of example 4 of the present invention;
FIG. 2 is a moisture profile of example 4 of the present invention;
figure 3 is the product of two treatments of example 4 of the present invention.
Detailed Description
The invention is further illustrated below with reference to the accompanying drawings and examples:
the experimental procedures in the following examples are conventional unless otherwise specified. The experimental materials used in the following examples were, unless otherwise specified, all of conventional biochemical preparations available on the market.
Example 1: preparing a first compound biological decomposition agent:
a. preparation of clostridium thermocellum pervone: (1) seed culture: inoculating the bacterial colony on the plate culture medium into a seed culture solution, and performing shake cultivation at 55-60 ℃ for 16-24 h at the rotating speed of 200-260 rpm;
wherein, the seed culture solution is: 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water, wherein the pH value is adjusted to be 6.6-7.0; 1.5kg of the compound is prepared according to the mixture ratio.
(2) And (3) amplification culture: inoculating the seed culture solution into the expanded culture solution by an inoculation amount of 1.5%, and performing shake culture at 55-60 ℃ for 16-24 h at a rotation speed of 200-260 rpm;
wherein the amplification culture solution comprises 0.01g of formaldehyde, 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water, the balance of the solution is 100g, and the pH value is adjusted to be 6.6-7.0; 100kg of expanded culture solution is prepared according to the proportion for use.
(3) Preparation of a microbial inoculum: adding 0.1kg of protective agent and 30kg of carrier adsorbent into the product cultured in the step (2), and fully drying to obtain a powdery microbial inoculum;
wherein the protective agent is soybean protein and whey powder with the weight ratio of 1:1, and the carrier adsorbent is talcum powder.
b. B, preparing the clostridium thermophilum chufa bacterial agent prepared in the step a and other materials according to the following parts by weight: 5 parts of clostridium paphiolyticum, 2 parts of lactobacillus thermophilus, 20 parts of composite bacteria (the weight ratio of bacillus subtilis, pseudomonas and bacillus pumilus is 8:1:1), 2 parts of a composite enzyme preparation (the weight ratio of acid protease, endo-cellulase and xylanase is 5:1:1), 1 part of composite actinomycetes (the weight ratio of pseudomonas parvum, streptomyces michiganensis and streptomyces rochei is 2:1:1), 5 parts of composite fungi (the weight ratio of trichoderma viride and aspergillus niger is 1:3), 65 parts of an organic carrier (rice hull powder) and 0.001 part of tea tree essence.
Adding the clostridium paphioparum inoculum into a prepared organic carrier in proportion, and regulating the rotating speed to 150rmp under the condition of continuous stirring; sequentially adding Lactobacillus thermophilus, composite bacteria, composite enzyme preparation, composite actinomycetes and composite fungi, and stirring for at least 5min when adding one genus; spraying 0.001 wt% of tea tree essential oil while adding each thallus, and stirring thoroughly for adsorption.
Example 2: preparing a second compound biological decomposition agent:
a. preparation of clostridium thermocellum pervone: (1) seed culture and (2) expansion culture were the same as in example 1.
(3) Preparation of a microbial inoculum: adding 0.2kg of protective agent and 25kg of carrier adsorbent into the product cultured in the step (2), and fully drying to obtain a powdery microbial inoculum;
wherein the protective agent is soybean protein and whey powder in a weight ratio of 3:1, and the carrier adsorbent is beneficial powder.
b. B, preparing the clostridium thermophilum chufa bacterial agent prepared in the step a and other materials according to the following parts by weight: 5 parts of clostridium papyrhizae, 2 parts of lactobacillus thermophilus, 20 parts of composite bacteria (the weight ratio of bacillus subtilis, pseudomonas and bacillus pumilus is 8:1:1), 2 parts of a composite enzyme preparation (the weight ratio of acid protease, endo-cellulase and xylanase is 5:1:1), 1 part of composite actinomycetes (the weight ratio of pseudomonas parvum, streptomyces micheli and streptomyces rochei is 2:1:1), 5 parts of composite fungi (the weight ratio of trichoderma viride to aspergillus niger is 1:3), 60 parts of an organic carrier (rice hull powder) and 0.001 part of tea tree essential oil.
Under the condition of continuous stirring, the rotating speed is adjusted to be 150rmp, the clostridium thermocellum microbial inoculum is added into a prepared organic carrier in proportion, and tea tree essential oil is sprayed at the same time; sequentially adding Lactobacillus thermophilus, composite bacteria, composite enzyme preparation, composite actinomycetes and composite fungi, and stirring for at least 5min when adding one strain; and in the process of adding each thallus, spraying 0.001 wt% of tea tree essential oil liquid, and fully stirring and adsorbing to obtain the tea tree essential oil.
Example 3: preparation control of Clostridium paphioparvum
Referring to the preparation method of example one, the induction was performed without adding formaldehyde to the expanded culture medium (uninduced group). In the microbial inoculum obtained under the same preparation conditions, the content of the clostridium thermocellum liquid for induction (uninduced group) without adding formaldehyde is 35 hundred million/ml, and the content of the clostridium thermocellum liquid for induction (induced group) with adding formaldehyde is as high as 82 hundred million/ml. Cellulose degradation experiments of reed straws by using the clostridium thermophilum prepared by the two methods are compared.
The experimental method comprises the following steps: 1. compounding 50% of reed straw powder, 10% of rice bran, 34% of rice hull vinasse, 4% of corn flour, 1% of urea, 0.4% of yeast powder and 0.3% of potassium dihydrogen carbonate into a solid culture medium, sterilizing at the high temperature of 121 ℃ for 30min for later use, and measuring the crude fiber content of the culture medium. 2. The clostridium thermocellum peruvianum prepared by the two methods (uninduced group and induced group) is respectively treated according to the following ratio of 1: 100, uniformly spraying the mixture to a solid culture medium prepared from reed straw powder, making a blank control, and adjusting the dry-wet ratio. 3. Culturing at 30 deg.C for 72h, and turning over the pile 2-3 times daily. The experimental data are detailed in table 1:
TABLE 1 comparison of the effect of Clostridium thermocellum on the degradation of crude fiber
| Treatment of | Dried and then weighed (g) | Ashed weight (g) | Crude fiber (%) | Rate of degradation |
| Blank space | 27.6623 | 27.2072 | 42.39％ | |
| Uninduced group | 27.1344 | 27.0125 | 29.32％ | 30.83％ |
| Induction group | 27.3267 | 27.0345 | 23.27％ | 45.10％ |
Therefore, the clostridium thermocellum perusal induced by formaldehyde has higher yield, better activity and stronger cellulose decomposition capability.
Example 4: application of composite biological decomposition agent
In 8 months of 2020, the composite biological decomposition agent (treatment 2) prepared in example 1 and a decomposition agent with a better market effect (organic material decomposition agent of Jiangsu Green technology Co., Ltd. is used as a comparison treatment 1, and the main components are Bacillus subtilis, Bacillus licheniformis, yeast, mold and the like and metabolites thereof), are compared and tested, the mixed material of sludge and straw according to the volume ratio of 1:0.7 is decomposed, and the test treatment is shown in table 2, table 3, figure 1, figure 2 and figure 3:
TABLE 2 market price of decomposition agent, decomposed material and amount
| Treatment of | Decomposed material | Dosage of | Price |
| Process 1 | Sludge: the ratio of the straw to the straw is 1:0.7 | 1kg/10 ton | Market price of 30000 yuan/ton |
| Treatment 2 | Sludge: the ratio of the straw to the straw is 1:0.7 | 1kg/10 ton | Production cost of 9700 yuan/ton |
TABLE 3 physical, chemical and biological index of the decomposed material
As can be seen from figure 1, the initial temperature of each treatment mixture is 40.5 ℃, the treatment 2 reaches 62.33 ℃ in 10 days in 8 months and enters the medium-high temperature region, the duration is 12 days, the maximum fermentation temperature is 69.44 ℃, the high temperature duration is long due to quick temperature rise, the temperature is reduced to 45.22 ℃ in 17 days in 8 months, and the fermentation is finished in 29 days in 8 months. While the treatment 1 reached 60.10 ℃ in 8/16 th day, entered the high temperature zone for 5 days, and then entered the secondary high temperature zone, the temperature did not decrease for a long time due to incomplete fermentation, and the temperature was 55.10 ℃ in 8/29 th day, and the temperature approached the initial temperature after one week. Therefore, the treatment 2 is superior to the treatment 1 in view of the speed and amplitude of the starting temperature and the duration of the high temperature, and the whole fermentation period of the treatment 2 is shortened by one week compared with the treatment 1, so that the fermentation is more complete.
As can be seen from FIG. 2, the initial water content in each treatment room was 1: 72.21% and 2: 72.48%, the water content in the mixed material was too high and 70% or more, the temperature was difficult to rise, and the decomposition rate was significantly reduced, because too much water filled the compost material particles with water, which obstructed ventilation, thereby causing anaerobic conditions, and H was generated2S, which is not beneficial to the growth of aerobic microorganisms. However, the final water contents of treatments 1 and 2 were 61.67% and 54.33%, respectively, and the water reductions were 14.60% and 25.04%, respectively, from the comparison of the treatments, and treatment 2 was superior to treatment 1.
As can be seen from Table 2, Table 3 and FIG. 3, the product of the present invention has low cost, high profit and good effect of the decomposed material obtained by processing the same material as the comparison.
It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.
Claims (5)
1. The preparation method of the composite biological decomposition agent is characterized in that the composite biological decomposition agent consists of the following components in parts by weight: 2-10 parts of clostridium papulophilus, 1-5 parts of lactobacillus thermophilus, 10-30 parts of composite bacteria, 2-5 parts of a complex enzyme preparation, 2-5 parts of composite actinomycetes, 5-10 parts of composite fungi, 40-70 parts of an organic carrier and 0.001 part of tea tree essential oil;
the compound bacteria comprise one or more of bacillus subtilis, bacillus amyloliquefaciens, pseudomonas and bacillus pumilus; the compound enzyme preparation comprises one or more of acid protease, endo-cellulase and xylanase; the compound actinomycetes comprises one or more of Mycobacterium, Micromonospora, Streptomyces melanioides and Streptomyces rochei; the composite fungus comprises one or more of Trichoderma viride, Trichoderma reesei and Aspergillus niger; the organic carrier comprises one or more of rice hull powder, bran and straw powder;
the preparation method of the composite biological decomposition agent comprises the following steps:
a. preparation of clostridium paphiolyticum: (1) seed culture: inoculating the bacterial colony on the plate culture medium into a seed culture solution, and performing shake cultivation at 55-60 ℃ for 16-24 h at the rotating speed of 200-260 rpm;
wherein, the seed culture solution is: 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water, wherein the pH value is adjusted to be 6.6-7.0;
(2) and (3) amplification culture: inoculating the seed culture solution into the expanded culture solution in an inoculation amount of 1-5%, and performing shake cultivation at 55-60 ℃ for 16-24 h at a rotation speed of 200-260 rpm;
wherein the expanding culture solution is 0.01g of formaldehyde, 50g of cellobiose solution, 1g of tryptone, 0.5g of yeast extract, 0.15g of ammonium chloride, 0.5g of urea, 0.2g of tripotassium citrate, 0.15g of citric acid monohydrate, 0.1g of monopotassium phosphate, 0.1g of sodium chloride and water for supplementing 100g, and the pH value is adjusted to be 6.6-7.0;
(3) preparation of a microbial inoculum: adding 0.1-0.5% by weight of protective agent and 10-30% by weight of carrier adsorbent into the product cultured in the step (2), and fully drying to obtain a powdery microbial inoculum;
b. and (b) adding the clostridium thermophilum agent of the cyperus alternifolius prepared in the step a into a prepared organic carrier according to a proportion, sequentially and respectively adding lactobacillus thermophilus, composite bacteria, a composite enzyme preparation, composite actinomycetes and composite fungi under the condition of continuous stirring, spraying 0.001 weight percent of tea tree essential oil solution in the process of adding all the bacteria and uniformly mixing, and fully stirring and adsorbing to obtain the cyperus alternifolius microbial inoculum.
2. The method for preparing a composite biological decomposition agent according to claim 1, wherein: the weight parts of the clostridium papyrus, the lactobacillus thermophilus, the composite bacteria, the composite enzyme preparation, the composite actinomycetes, the composite fungi, the organic carrier and the tea tree essential oil are 5 parts, 2 parts, 20 parts, 2 parts, 1 part, 5 parts, 65 parts and 0.001 part respectively.
3. The method for preparing a composite biological decomposition agent according to claim 1, wherein: the protective agent is one or more of skimmed milk, soybean protein and whey powder.
4. The method for preparing a composite biological decomposition agent according to claim 1, wherein: the carrier adsorbent is one or more of talcum powder, light calcium carbonate and beneficial powder.
5. The method for preparing a composite biological decomposition agent according to claim 1, wherein: and in the step b, the stirring speed is 100-200 rmp, and each group of bacteria is stirred for more than 5min during adding.
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