A method for preparing solid decomposing bacteria agent

Abstract

The present invention discloses a method for preparing a solid decomposing agent, which comprises the following steps: a first step, making a solid fermentation device; a second step, preparing a seed liquid; a third step, preparing a fermentation liquid; and a fourth step, solid fermentation to obtain a solid decomposing agent. The beneficial effects of the present invention are as follows: 1. A mixture of bacteria and fungi is used in the product, and the spores formed after fungal fermentation are not easy to germinate, and the bacterial content after bacterial fermentation is relatively stable, and the shelf life is longer than that of ordinary products. 2. The experiment targets the main components of the fermentation materials, such as cellulose, protein, starch and lignin, to screen the fermentation functional strains, and the product has the function of decomposing cellulose, protein, starch and lignin, and the product has a wide range of uses. 3. The experiment develops a special solid fermentation formula to increase the bacterial content in the unit product, thereby reducing the amount of the product used and reducing the decomposition cost.

Description

Technical Field

[0001] The invention relates to the field of decomposing agents, and in particular to a preparation method of a solid decomposing bacterial agent. Background Art

[0002] With the development of agricultural planting, large-scale planting of crops has gradually occupied a major position in the industry. In order to meet the demand for agricultural and sideline products due to the large population base in my country, large-scale planting of crops relies on the input of a large amount of chemical fertilizers. With the increase in the amount of chemical fertilizers, the pressure on the surrounding natural ecological environment and soil environment has gradually increased. In response to changes in the planting environment, the country has proposed a series of policies to reduce the application of chemical fertilizers and increase the application of organic fertilizers. With the promotion of national policies, the application of organic fertilizers in agricultural planting will gradually occupy a dominant position. In the process of organic fertilizer production, fermentation is the key to determining the quality of organic fertilizers. In traditional agriculture, the application of straw return to the field, straw burning and (animal and plant waste) pile return to the field and other models have failed to reflect the application value of organic fertilizers. The main reason is that the materials are not fully decomposed when applied, and burning will pollute the air and the surrounding environment. Incineration is also a waste of nutrient resources of straw materials. Direct return to the field or pile return to the field has the risk of affecting the next season's crops. Fully decomposing and fermenting organic materials has become the focus of the promotion and application of organic fertilizers. Summary of the invention

[0003] The purpose of the present invention is to provide a method for preparing a solid decomposition bacteria agent in view of the deficiencies in the prior art.

[0004] To achieve the above object, the present invention is implemented through the following technical solutions.

[0005] A method for preparing a solid decomposing bacteria agent comprises the following steps:

[0006] The first step is to make a solid fermentation device. The cover of a gasoline barrel (1) with a diameter of 40 cm and a height of 120 cm is cut off by oxygen welding. A hole (2) is punched at a position 10 cm below the bottom of the gasoline barrel. The hole diameter is 7 cm. A steel bar with a width of 2 cm and a thickness of 0.2 cm is welded at a position 5 cm square to the top of the hole. One steel bar is welded every 5 cm, and a total of 8 steel bars are welded. After welding, a sieve (3) with an aperture of 2 mm is attached. The hole (2) is connected by a hose. The other end of the hose is connected to a blower (4). A heating device (5) is provided at the air inlet of the blower (4).

[0007] The second step is to prepare the seed solution. The four preserved functional bacteria: functional bacteria A, functional bacteria B, functional bacteria C, and functional fungus D are activated respectively. Functional bacteria A, functional bacteria B, and functional bacteria C are cultured using LB basic plate culture medium, and functional fungus D is cultured using PDA basic plate culture medium. After activation, functional bacteria A, functional bacteria B, and functional bacteria C are inoculated into 200 mL of liquid LB culture medium respectively, and cultured in an incubator at a temperature of 35°C and a rotation speed of 180 r / min for 48 hours. The cultured liquid LB culture medium is added into 1000 mL of LB liquid culture medium, and the cultured liquid LB culture medium is added into an incubator at a temperature of 35°C and a rotation speed of 180 r / min for 48 hours. Functional bacteria D is inoculated into 200 mL of liquid PDA culture medium, and cultured in an incubator at a temperature of 28°C and a rotation speed of 180 r / min for 72 hours. The cultured liquid PDA culture medium is added into 1000 mL of The PDA liquid medium was placed in an incubator at 28°C and 180 r / min for 72 hours to prepare seed solution A, seed solution B, seed solution C and seed solution D respectively;

[0008] The third step is to prepare the fermentation broth. 600 L of fermentation medium was added to four fermentation tanks with a volume of 1000 L. The four different seed solutions were added to the four fermentation tanks respectively. 3 / h, temperature 28 ° C and rotation speed 160r / min, fermentation for 48h, to prepare fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D;

[0009] The fourth step is solid fermentation. The fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D are mixed, and the mixed fermentation liquid is added to the solid culture medium at a volume ratio of 1:10. Water is added to adjust the water content of the fermentation material to 60% to 70%, and the mixture is mixed evenly in a mixer. After mixing, the fermentation material is loaded into the fermentation device prepared in the first step. The height of the mixed material is 80 cm to 90 cm, and the temperature is 28 ° C to 30 ° C and the ventilation volume is 5m 3 / h for 60h. After 60h, at 30℃~35℃ and ventilation volume 5m 3 / h conditions for fermentation for 36 hours. After 36 hours, the mixed material is added into a drying barrel and dried at a temperature of 50°C to 55°C until the water content of the material drops to 10% to 15% (mass ratio). After drying, the material is crushed to obtain a solid composting agent.

[0010] Furthermore, the functional strain A in the second step is Bacillus subtilis, with a deposit number of CGMCC NO.15844;

[0011] The functional strain B is Bacillus stearothermophilus, and its deposit number is CGMCC NO.15852;

[0012] The functional strain C is Bacillus licheniformis, and its deposit number is CGMCCNO.20149;

[0013] The functional strain D shown is Aspergillus oryzae, and its deposit number is CGMCC NO.23280.

[0014] Furthermore, the composition of the fermentation medium in the third step is 3g / L-5g / L yeast extract, 3g / L-5g / L peptone, 8g / L-10g / L corn steep liquor powder, 3g / L-5g / L corn starch, 0.2g / L-0.5g / L dipotassium hydrogen phosphate, 0.1g / L-0.3g / L potassium dihydrogen phosphate, 10g / L-15g / L glucose and 1g / L-10g / L molasses powder.

[0015] Furthermore, in the fourth step, the fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D are mixed in a mass ratio of 1.5:1.5:0.5:0.5.

[0016] Furthermore, the formula of the solid culture medium in the fourth step is 25.0g / L-30.0g / L of bran, 30.0g / L-35g / L of sawdust, 17.0g / L-22.0g / L of rice husk, 10.0g / L-15g / L of corn flour, 4.2g / L-5.2g / L of (NH4)2SO4, 2.0g / L-3.0g / L of glucose, 0.5g / L-1.0g / L of K2HPO4, and 0.3g / L-0.5g / L of MgSO4·7H2O.

[0017] Beneficial effects of the present invention:

[0018] 1. The product uses a mixture of bacteria and fungi. After the fungi are fermented, the spores are not easy to germinate. After the bacteria are fermented, the bacterial content is relatively stable, and the shelf life is longer than that of ordinary products.

[0019] 2. The experiment targeted the main components of the fermentation materials, such as cellulose, protein, starch and lignin, to screen the fermentation functional strains. The product has the function of decomposing cellulose, protein, starch and lignin, and has a wide range of applications.

[0020] 3. Experimentally develop special solid fermentation formulas to increase the bacterial content per unit product, thereby reducing the amount of product used and reducing composting costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 It is a structural schematic diagram of the solid fermentation device of the present invention;

[0022] Figure 2 This is a schematic diagram showing the effect of different decomposing bacteria on the fermentation temperature of oil cake;

[0023] In the figure: 1. gasoline barrel; 2. holes; 3. screen; 4. blower; 5. heating device. DETAILED DESCRIPTION

[0024] The present invention will be further described below in conjunction with the embodiments, but is not limited to the contents of the description.

[0025] A method for preparing a solid decomposing bacterial agent comprises the following steps:

[0026] The first step is to make a solid fermentation device. The lid of a gasoline barrel 1 with a diameter of 40 cm and a height of 120 cm is cut off by oxygen welding. A hole 2 with a diameter of 7 cm is drilled at a position 10 cm below the bottom of the gasoline barrel. A steel bar with a width of 2 cm and a thickness of 0.2 cm is welded 5 cm above the hole. One steel bar is welded every 5 cm, for a total of 8 steel bars. After welding, a screen 3 with an aperture of 2 mm is attached. The hole is connected with a hose, and the other end of the hose is connected to a blower 4. At the same time, a heating device 5 is placed at the air inlet of the blower.

[0027] The second step is the preparation of seed solution. The four preserved functional bacteria (functional bacteria A, functional bacteria B, functional bacteria C, and functional fungus D) are activated respectively. Functional bacteria A, functional bacteria B, and functional bacteria C are cultured using LB basic plate culture medium, and functional fungus D is cultured using PDA basic plate culture medium. After activation, functional bacteria A, functional bacteria B, and functional bacteria C are inoculated into 200 mL of liquid LB culture medium respectively, placed in an incubator at a temperature of 35°C and a rotation speed of 180 r / min for 48 hours, and then the cultured liquid LB culture medium is added into 1000 mL of LB liquid culture medium, placed in an incubator at a temperature of 35°C and a rotation speed of 180 r / min for propagation for 48 hours, and functional bacteria D is inoculated into 200 mL of liquid PDA culture medium, placed in an incubator at a temperature of 28°C and a rotation speed of 180 r / min for 72 hours, and then the cultured liquid PDA culture medium is added into 1000 mL of The seeds were placed in PDA liquid culture medium in an incubator at 28°C and 180 r / min for 72 hours to prepare seed solution A, seed solution B, seed solution C and seed solution D respectively.

[0028] The third step is to prepare the fermentation liquid. 600L of special fermentation medium A was added to four fermentation tanks with a volume of 1000L. The four different seed liquids were added to the four fermentation tanks respectively. 3 / h, temperature 28 ° C and rotation speed 160r / min, and fermented for 48h to prepare fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D.

[0029] The fourth step is solid fermentation. The fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D are mixed at a ratio of 1.5:1.5:0.5:0.5, and the mixed fermentation liquid is added to the solid culture medium E at a volume ratio of 1:10. Water is added to adjust the water content of the fermentation material to 60% to 70%, and the mixture is mixed evenly in a blender. After mixing, the fermentation material is loaded into the fermentation device, and the height of the mixed material is 80 cm to 90 cm. The temperature is 28°C to 30°C and the ventilation volume is 5m 3 / h for 60h. After 60h, at 30℃~35℃ and ventilation volume 5m 3 / h conditions for fermentation for 36 hours. After 36 hours, the mixed material is added into a drying barrel and dried at a temperature of 50°C to 55°C until the water content of the material drops to 10% to 15% (mass ratio). After drying, the material is crushed to obtain a solid composting agent.

[0030] The functional strain A is Bacillus subtilis, with a deposit number of CGMCC NO.15844, and a depository unit: General Microbiology Center of China Culture Collection Administration; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is June 4, 2018.

[0031] The functional strain B is Bacillus stearothermophilus, with a deposit number of CGMCC NO.15852, and a depository unit: General Microbiology Center of China Culture Collection Administration; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is June 4, 2018.

[0032] The functional strain C is Bacillus licheniformis, with a deposit number of CGMCCNO.20149. The depository unit is the General Microbiology Center of China Culture Collection Administration; the deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit date is June 28, 2020.

[0033] The functional strain D is Aspergillus oryzae, with a preservation number of CGMCC NO.23280. The preservation unit is the General Microbiology Center of China Culture Collection Administration; the preservation address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the preservation date is October 18, 2021.

[0034] The formula of the liquid special culture medium A in the third step is 3g / L~5g / L yeast extract, 3g / L~5g / L peptone, 8g / L~10g / L corn steep liquor powder, 3g / L~5g / L corn starch, 0.2g / L~0.5g / L dipotassium hydrogen phosphate, 0.1g / L~0.3g / L potassium dihydrogen phosphate, 10g / L~15g / L glucose and 1g / L~10g / L molasses powder.

[0035] The formula of solid culture medium E is 25.0g / L~30.0g / L bran, 30.0g / L~35g / L sawdust, 17.0g / L~22.0g / L rice husk, 10.0g / L~15g / L corn flour, 4.2g / L~5.2g / L (NH4)2SO4, 2.0g / L~3.0g / L glucose, 0.5g / L~1.0g / L K2HPO4, and 0.3g / L~0.5g / L MgSO4·7H2O.

[0036] Product effect verification:

[0037] 1. Effect of solid decomposition bacteria on fermentation with oil cake as main raw material

[0038] Through fermentation contrast tests, the effect of optimized decomposing bacteria on fermented organic fertilizer with oil cake as the main raw material was verified, aiming to form the technology for using solid decomposing bacteria in fermented oil cake organic fertilizer.

[0039] 1.1 Materials and methods

[0040] 1.1.1 Test location

[0041] The experiment was arranged in Qinglong, Anning.

[0042] 1.1.2 Test materials

[0043] Tested composting agents: solid composting agent, conventional composting agent on the market, and original composting agent.

[0044] Test materials: oil cake, peat.

[0045] 1.1.3 Test methods

[0046] 1.1.3.1 Experimental design

[0047] The experiment set up 4 treatments, each of which fermented 200 kg of materials. The experimental treatments are as follows:

[0048] Treatment 1: Add 5% solid compost during fermentation;

[0049] Treatment 2: Add 5% of conventional composting agent on the market (straw solid composting agent produced by Shandong Beijia Biotechnology Co., Ltd.) during fermentation;

[0050] Treatment 3: Add 5% of the original composting agent (liquid composting agent for straw produced by Yunnan Microbial Fermentation Engineering Research Center Co., Ltd.) during fermentation;

[0051] Treatment 4: No composting agent added;

[0052] Except for the different weights of composting agents used in the experiment, other composting and fermentation conditions remained the same.

[0053] 1.1.3.2 Test operation

[0054] (1) Raw material crushing

[0055] The raw materials for each treatment were pulverized by a pulverizer and then sieved through a 20-mesh sieve.

[0056] (2) Mixed fermentation of raw materials

[0057] Mix the raw materials after each treatment and screening evenly, then add water and stir evenly to ensure that the moisture content of the material is about 50% (it can be basically dispersed after being pinched into a ball by hand and dropped to the ground with force). At the same time, ensure that the quantity of the material is basically accurate. If the moisture content of the material is high, it should be dried first to reduce the moisture content, and then piled into a cone shape for fermentation.

[0058] (3) Temperature measurement and compost turning

[0059] Every day, measure and record the material temperature at the upper, middle and lower layers of the material pile, and also measure and record the ambient temperature at that time. Turn the pile three hours after the temperature reaches above 40°C, and turn it once a day until the temperature approaches the ambient temperature. If the temperature is still below 40°C after 48 hours, turn the pile to avoid creating an anaerobic environment; in the middle and late stages of fermentation, if the temperature reaches above 60°C, turn the pile to avoid excessive temperature and killing beneficial bacteria.

[0060] 1.1.4 Investigation and Recording

[0061] 1.1.4.1 Determination of temperature changes

[0062] The temperature of each pile is measured at 10 a.m. and recorded. At the same time, the changes in the pile, such as changes in odor, mycelium growth, whether the fertilizer is clumping, etc., are observed and corresponding records are made.

[0063] 1.1.4.2 Determination of nutrients in organic fertilizer after fermentation

[0064] Take 2kg of sample and conduct a full inspection of the organic fertilizer according to the determination method of the industry standard NY 525-2012 "Organic Fertilizer" to analyze the effects of different composting bacteria on material fermentation.

[0065] 1.1.4.3 Determination of seed germination index (GI)

[0066] (1) The samples collected on the day of fermentation and after fermentation were used to determine the seed germination rate.

[0067] (2) Air-dry the compost sample and take 5g of it into a 200mL beaker. Add 100mL of warm water at 60℃ and soak for 3h. Filter it. Take 10mL of the filtered juice and pour it into a Petri dish covered with two layers of filter paper. Sow 50 cabbage or radish seeds and carry out the germination test. A blank control is set up. Distilled water is used in the Petri dish. The seeds and germination test methods are the same as above.

[0068] (3) Collect samples according to the treatment and set up three replicates, and observe the results.

[0069] 2. Results and Analysis

[0070] 2.1 Effects of different fermentation agents on oil cake fermentation temperature

[0071] like Figure 2 As shown,

[0072] 2.2 Effects of different composting agents on the nutrients of oil cake organic fertilizer

[0073]

[0074]

[0075] 2.3 Determination of seed germination rate of different composting agents

[0076] deal with Germination rate Germination Index Process 1 96.00％ 124.69 Process 2 90.66％ 112.08 Process 3 91.34％ 115.78 Process 4 60.00％ 42.25

[0077] It can be seen from the above table that the composting agent of the present application has a great promoting effect on the germination rate of seeds, and the effect is better than that of commercially available products.

[0078] Obviously, the above-mentioned embodiments of the present invention are merely examples for clearly explaining the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is impossible to list all the embodiments here. Any obvious changes or modifications derived from the technical solution of the present invention are still within the scope of protection of the present invention.

Claims

1. A method for preparing a solid decomposing bacteria agent, characterized in that: The invention comprises the following steps: the first step is to prepare a solid fermentation device, wherein the cover of a gasoline barrel (1) with a diameter of 40 cm and a height of 120 cm is cut off by oxygen welding, a hole (2) is punched at a position 10 cm below the bottom of the gasoline barrel side, the hole diameter is 7 cm, a steel bar with a width of 2 cm and a thickness of 0.2 cm is welded at a position 5 cm square to the top of the hole, one steel bar is welded at a distance of 5 cm, and 8 steel bars are welded in total. After welding, a screen (3) with a hole diameter of 2 mm is attached, the hole (2) is connected by a hose, the other end of the hose is connected to a blower (4), and a heating device (5) is arranged at the air inlet of the blower (4); The second step is to prepare the seed solution. The four preserved functional bacteria: functional bacteria A, functional bacteria B, functional bacteria C, and functional fungus D are activated respectively. Functional bacteria A, functional bacteria B, and functional bacteria C are cultured using LB basic plate culture medium, and functional fungus D is cultured using PDA basic plate culture medium. After activation, functional bacteria A, functional bacteria B, and functional bacteria C are inoculated into 200 mL of liquid LB culture medium respectively, and cultured in an incubator at a temperature of 35°C and a speed of 180 r / min for 48 hours. The cultured liquid LB culture medium is then added to 1000 mL of LB culture medium. B liquid culture medium, placed in an incubator at a temperature of 35°C and a rotation speed of 180r / min for propagation for 48h, functional bacteria D was inoculated into 200mL liquid PDA culture medium, placed in an incubator at a temperature of 28°C and a rotation speed of 180r / min for 72h, the cultured liquid PDA culture medium was added to 1000mL PDA liquid culture medium, placed in an incubator at a temperature of 28°C and a rotation speed of 180r / min for propagation for 72h, and seed solution A, seed solution B, seed solution C and seed solution D were prepared respectively; The third step is to prepare the fermentation broth. 600 L of fermentation medium was added to four fermentation tanks with a volume of 1000 L. The four different seed solutions were added to the four fermentation tanks respectively. 3 / h, temperature 28 ° C and rotation speed 160r / min, fermentation for 48h, to prepare fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D; The fourth step is solid fermentation. The fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D are mixed, and the mixed fermentation liquid is added to the solid culture medium at a volume ratio of 1:

10. Water is added to adjust the water content of the fermentation material to 60% to 70%, and the mixture is mixed evenly in a mixer. After mixing, the fermentation material is loaded into the fermentation device prepared in the first step. The height of the mixed material is 80 cm to 90 cm, and the temperature is 28 ° C to 30 ° C and the ventilation volume is 5m 3 / h for 60h. After 60h, at 30℃~35℃ and ventilation volume 5m 3 / h conditions for fermentation for 36 hours, after 36 hours, add the mixed material into a drying barrel, dry it at a temperature of 50°C to 55°C until the moisture content of the material drops to 10% to 15% (mass ratio), and then grind it after drying to obtain a solid composting agent; In the second step, the functional strain A is Bacillus subtilis, with a deposit number of CGMCCNO.15844; The functional strain B is Bacillus stearothermophilus, with a deposit number of CGMCCNO.15852; The functional strain C is Bacillus licheniformis, and its deposit number is CGMCCNO.20149; The functional strain D shown is Aspergillus oryzae, and its deposit number is CGMCCNO.23280.

2. The method for preparing a solid decomposition agent according to claim 1, characterized in that: The fermentation medium in the third step is composed of 3g / L-5g / L yeast extract, 3g / L-5g / L peptone, 8g / L-10g / L corn steep liquor powder, 3g / L-5g / L corn starch, 0.2g / L-0.5g / L dipotassium hydrogen phosphate, 0.1g / L-0.3g / L potassium dihydrogen phosphate, 10g / L-15g / L glucose and 1g / L-10g / L molasses powder.

3. The method for preparing a solid decomposition agent according to claim 1, characterized in that: In the fourth step, the mass ratio of the mixture of fermentation liquid A, fermentation liquid B, fermentation liquid C and fermentation liquid D is 1.5:1.5:0.5:0.

5.

4. The method for preparing a solid decomposition agent according to claim 1, characterized in that: The formula of the solid culture medium in the fourth step is 25.0g / L-30.0g / L of bran, 30.0g / L-35g / L of sawdust, 17.0g / L-22.0g / L of rice husk, 10.0g / L-15g / L of corn flour, 4.2g / L-5.2g / L of (NH4)2SO4, 2.0g / L-3.0g / L of glucose, 0.5g / L-1.0g / L of K2HPO4, and 0.3g / L-0.5g / L of MgSO4·7H2O.

Images