CN112852666A - Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent - Google Patents

Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent Download PDF

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CN112852666A
CN112852666A CN202110066033.6A CN202110066033A CN112852666A CN 112852666 A CN112852666 A CN 112852666A CN 202110066033 A CN202110066033 A CN 202110066033A CN 112852666 A CN112852666 A CN 112852666A
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microbial
microbial agent
carrier
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石泉
王宏志
董亮
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Xinjiang Herun Water Industry Co ltd
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    • C12N1/20—Bacteria; Culture media therefor
    • 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
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • 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
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Abstract

The invention discloses a preparation method of a microbial agent and a microbial fertilizer prepared by the microbial agent. The method comprises the following steps: s1, drying, crushing and sieving plant straws to obtain a carrier; s2, mixing the carrier with alkaline water to obtain a treated carrier; s3, mixing the processed carrier with microbial cells or bacterial liquid containing the microbial cells; s4, adding a protective agent; s5, centrifuging or filtering to obtain the microbial agent. The microbial agent obtained by the preparation method of the invention takes plant straws treated by weak base as a carrier, and propylene glycol and carboxymethyl cellulose are used for protection treatment, so that the high temperature resistance of the microbial agent can be obviously improved, the growth inhibition and killing of microorganisms at high temperature in the composting process can be reduced, and the composting time can be shortened.

Description

Preparation method of microbial agent and microbial fertilizer prepared by adopting microbial agent
Technical Field
The invention belongs to the technical field of biological fertilizers, and particularly relates to a preparation method of a microbial agent and a microbial fertilizer prepared by the microbial agent.
Background
The microbial organic fertilizer is an organic material prepared by using animal and plant residues such as livestock and poultry excrement, crop straws and the like as raw materials through harmless treatment and decomposition, and a fertilizer product compounded with effective bacteria, and is a fertilizer with the effects of a microbial agent and an organic fertilizer.
The microbial agent for preparing the microbial fertilizer plays an important role in the preparation of the fertilizer. In the process of preparing the microbial fertilizer, the non-high temperature resistance of some microbial agents can be reduced along with the temperature rise, and the effective distribution of the subsequent components is influenced. In the existing preparation method of the microbial agent, the high temperature resistance of the microbial agent is specially researched less.
Disclosure of Invention
Aiming at the problems, the invention discloses a preparation method of a microbial agent and a microbial fertilizer prepared by the microbial agent. The high temperature resistance of the microbial agent prepared by the method is obviously higher than that of a common liquid microbial agent.
The first aspect of the invention discloses a preparation method of a microbial agent, which comprises the following steps:
s1, drying, crushing and sieving plant straws to obtain a carrier;
s2, mixing the carrier with alkaline water to obtain a treated carrier;
s3, mixing the processed carrier with microbial cells or bacterial liquid containing the microbial cells;
s4, adding a protective agent;
s5, centrifuging or filtering to obtain the microbial agent.
In some embodiments of the invention, the straw is selected from one or more of corn stover, wheat straw, rice straw.
In some embodiments of the invention, the sieving is through a 10-30 mesh sieve.
In some embodiments of the invention, the pH of the alkaline water is 7.5 to 8.5, preferably 7.8 to 8.1.
In some embodiments of the invention, the weight ratio of the vector to the bacterial cells is (1-10): 1, preferably (3-5): 1.
in some embodiments of the invention, the protectant is a mixture of glycerol and carboxymethylcellulose
In some embodiments of the invention, the concentration of glycerol in the system is 0.5-5g/ml, preferably 1.5-3 g/ml.
In some embodiments of the invention, the concentration of the carboxymethyl cellulose in the system is 5 to 15g/ml, preferably 8 to 12 g/ml.
In some embodiments of the invention, the microbial agent used as a starting material is a commercially available microbial agent comprising bacillus subtilis. Preferably, the bacillus subtilis is replaced by a composite strain comprising bacillus subtilis, wherein the composite strain comprises bacillus subtilis a, bacillus circulans, paenibacillus xylanolyticus, paenibacillus mucilaginosus and bacillus subtilis. The number ratio of the bacillus subtilis A to the bacillus circulans to the bacillus xylanilyticus to the bacillus mucilaginosus to the bacillus subtilis is (1-5): (1-5): (1-5): (1-5): (1-5).
The second aspect of the invention is to disclose a microbial fertilizer prepared by the microbial agent prepared by the preparation method of the first aspect.
The third aspect of the invention discloses a preparation method of the microbial fertilizer, which comprises the following steps:
s11, mixing animal or poultry excrement with straws and leaves to prepare a composting raw material;
s12, controlling the water content of the composting raw material between 50 and 55 percent;
s13, mixing and fermenting the microbial inoculum and the compost raw material with the regulated moisture, and turning the compost during the mixing and fermentation to control the fermentation temperature;
and S14, decomposing the raw materials to obtain the microbial fertilizer.
In some embodiments of the present invention, in S13, the amount of microbial agent added is determined by:
s131, using microbial fertilizer samples obtained in different addition amounts as arguments, U ═ x1, x2, …, Xn }, xi ═ (xi1, xi2, xi3, xi4), i ═ 1,2, …, n, using the color, odor, and fecal mass particle diameter of the microbial fertilizer samples as properties;
s132, standard deviation and range conversion is carried out through the following formulas:
Figure BDA0002904097870000021
Figure BDA0002904097870000022
is mean value, SkIs the standard deviation;
Figure BDA0002904097870000023
s133, order
Figure BDA0002904097870000024
Wherein C is a constant and takes a value of 0.38-0.40;
s134, obtaining a transfer closure matrix through a cubic flattening method, and classifying according to the size of lambda;
s135, if the lambda is larger than 0.41 and the materials are divided into at least two types, determining the addition amount in the type with the minimum addition amount as the alternative addition amount;
s136, if the lambda is less than 0.41, repeating S131-S134 until the lambda is more than 0.41;
and S137, repeating the steps for 3 times, and selecting the minimum candidate addition amount as the addition amount.
The beneficial technical effects of the invention are as follows:
the microbial agent obtained by the preparation method of the invention takes plant straws treated by weak base as a carrier, and propylene glycol and carboxymethyl cellulose are used for protection treatment, so that the high temperature resistance of the microbial agent can be obviously improved, the growth inhibition and killing of microorganisms at high temperature in the composting process can be reduced, and the composting time can be shortened.
The microbial fertilizer prepared by the microbial agent obtained by the preparation method has the carbon-nitrogen ratio and organic matter content which are in accordance with the standard of the compound organic fertilizer.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope 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. In the preparation methods of the microbial agents in the examples and the comparative examples, the added microbial agent is a commercial microbial agent sold in the market, the main strains are bacillus subtilis, lactic acid bacteria, saccharomycetes and photosynthetic bacteria, and the CFU ratio is approximately 1500:100:3: 1.
Example 1
The preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 30g of the mixture into 450ml of water, wherein the pH value of the water is 8.5, and stirring for 30 min;
adding microbial agent, and stirring for 5min to obtain wet thallus weight of 5 g;
adding 0.5g of glycerol and 3g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Example 2
The preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 70g of the mixture into 450ml of water, wherein the pH value of the water is 7.5, and stirring for 30 min;
adding microbial agent, and stirring for 5min, wherein the weight of the microbial agent is 10g in terms of wet weight of thallus;
adding 1.5g of glycerol and 7g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Example 3
The preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 50g of the mixture into 450ml of water, wherein the pH value of the water is 8.0, and stirring for 30 min;
adding microbial agent, and stirring for 5min, wherein the weight of the microbial agent is 10g in terms of wet weight of thallus;
adding 1g of glycerol and 5g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Example 4
The preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 10g of the mixture into 450ml of water, wherein the pH value of the water is 8.0, and stirring for 30 min;
adding microbial agent, and stirring for 5min, wherein the weight of the microbial agent is 10g in terms of wet weight of thallus;
adding 1g of glycerol and 5g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Example 5
The preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 50g of the mixture into 450ml of water, wherein the pH value of the water is 8.0, and stirring for 30 min;
adding microbial agent, and stirring for 5min, wherein the weight of the microbial agent is 20g in terms of wet weight of thallus;
adding 1g of glycerol and 5g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Example 6
The method for preparing the microbial fertilizer by using the microbial agent prepared in the examples 1-5 and the comparative examples 1 and 2 comprises the following steps:
mixing fresh pig manure and corn straws in a weight ratio of 1:1, and spraying water to adjust the water content to 50-55% to obtain a composting raw material;
taking the obtained microbial agent, converting into the microbial agent added in the preparation, mixing with the compost raw materials twice according to the weight ratio of 1:100, uniformly stirring, and preparing into a pile with the length, width and height of 20:1: 1;
the microbial inoculum can be directly mixed with the composting raw materials or dispersed by water and then mixed with the composting raw materials, and the water content of the last mixture is ensured to meet the requirement by mainly controlling the water content.
The pile was turned once every day for the first 30 days and then every 5 days.
Example 7
The method for preparing the microbial fertilizer by using the microbial agent prepared in the examples 1-5 and the comparative examples 1 and 2 comprises the following steps:
the difference from example 6 is that in the mixed fermentation of the microbial agent and the compost raw material with the adjusted moisture content, the amount of the microbial agent to be added is determined by the following method:
s131, using microbial fertilizer samples obtained in different addition amounts as arguments, U ═ x1, x2, …, Xn }, xi ═ (xi1, xi2, xi3, xi4), i ═ 1,2, …, n, using the color, odor, and fecal mass particle diameter of the microbial fertilizer samples as properties;
s132, standard deviation and range conversion is carried out through the following formulas:
Figure BDA0002904097870000051
Figure BDA0002904097870000052
is mean value, SkIs the standard deviation;
Figure BDA0002904097870000053
s133, order
Figure BDA0002904097870000054
Wherein, C is a constant and takes a value between 0.38 and 0.40;
s134, obtaining a transfer closure matrix through a cubic flattening method, and classifying according to the size of lambda;
s135, if the lambda is larger than 0.41 and the materials are divided into at least two types, determining the addition amount in the type with the minimum addition amount as the alternative addition amount;
s136, if the lambda is less than 0.41, repeating S131-S134 until the lambda is more than 0.41;
and S137, repeating the steps for 3 times, and selecting the minimum candidate addition amount as the addition amount.
In the process of preparing the microbial fertilizer by using the microbial agent, the addition amount of the microbial agent is a key control parameter. According to the preparation method, the color, the odor and the fecal mass particle diameter of the fertilizer sample are taken as properties, the addition amount of the microbial agent in the mixed fermentation of the microbial agent and the compost raw material for adjusting the moisture is determined by a fuzzy clustering analysis method, and the addition amount of the microbial agent can be rapidly determined by combining the fermentation time, so that the cost for using the microbial agent is reduced on the premise of ensuring the quality of the microbial fertilizer.
Comparative example 1
The difference from example 1 is that the preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving by a 20-mesh sieve;
adding 30g of the mixture into 450ml of water, wherein the pH value of the water is 8.5, and stirring for 30 min;
adding microbial agent, and stirring for 5min to obtain wet thallus weight of 5 g;
adding 0.5g propylene glycol and 3g chitin, and stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
Comparative example 2
The difference from example 1 is that the preparation method of the microbial agent comprises the following steps:
taking corn straws, drying, crushing and sieving with a 5-mesh sieve;
adding 30g of the mixture into 450ml of water, wherein the pH value of the water is 8.5, and stirring for 30 min;
adding microbial agent, and stirring for 5min to obtain wet thallus weight of 5 g;
adding 0.5g of glycerol and 3g of carboxymethyl cellulose, and continuing stirring for 30 min;
filtering or centrifuging to obtain the microbial agent.
High temperature resistance of microbial Agents prepared in Experimental example 1
Taking the microbial inoculum prepared in the examples and the comparative examples, adding 1ml of beef extract peptone liquid culture medium diluted by 10 times in terms of the wet weight of the thalli, and culturing for 10 days; washing with sterile water for 3 times, making into serial 10-fold dilutions, mixing with agar culture medium, culturing for 48 hr, and recording the number of colonies on the plate. The control was 5g of cells obtained by direct separation of microbial agents without treatment with corn stover, glycerol, and carboxymethylcellulose. The relative value of the number of colonies was set to 100% for example 1, i.e., the number of colonies/the number of colonies in example 1 × 100%. The results are shown in Table 1.
High temperature resistance of the microbial Agents prepared in Table 1
Relative value of colony counts/%)
Example 1 100e
Example 2 105e
Example 3 130f
Example 4 95d
Example 5 90d
Comparative example 1 60b
Comparative example 2 75c
Control 50a
The different lower case letters represent significant differences, P ═ 0.05
Experimental example 2
Microbial fertilizer prepared by using microbial agent prepared in examples 1-5 and comparative examples 1 and 2
The initial stage of composting has large odor, yellow brown color and large fecal mass, the total odor is reduced in the fermentation process, the color is blackened, and the diameter of the fecal mass is reduced. Until black brown, loose and no obvious stink.
The microorganisms of examples 1 to 5 showed no significant odor after about 60 days, and had a water content of about 30%, thereby completing the decomposition. Comparative example 1 is about 80 days and comparative example 2 is about 70 days.
The carbon nitrogen ratio of the decomposed organic fertilizer is about 18, the organic matter content is about 50%, and the finished product standard of the organic fertilizer is met.
While the preferred embodiments and examples of the present invention have been described in detail, the present invention is not limited to the embodiments and examples, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art.

Claims (10)

1. A preparation method of a microbial agent comprises the following steps:
s1, drying, crushing and sieving plant straws to obtain a carrier;
s2, mixing the carrier with alkaline water to obtain a treated carrier;
s3, mixing the processed carrier with microbial cells or bacterial liquid containing the microbial cells;
s4, adding a protective agent;
s5, centrifuging or filtering to obtain the microbial agent.
2. The method of claim 1, wherein the straw is selected from one or more of corn stover, wheat straw, rice straw.
3. The method of claim 1, wherein said sieving is through a 10-30 mesh sieve.
4. The method according to claim 1, characterized in that the pH of the alkaline water is 7.5-8.5, preferably 7.8-8.1.
5. The method according to claim 1, wherein the weight ratio of the carrier to the bacteria is (1-10): 1, preferably (3-5): 1.
6. the method of claim 1, wherein the protectant is a mixture of glycerol and carboxymethylcellulose.
7. The method according to claim 6, wherein the concentration of glycerol in the system is 0.5-5g/ml, preferably 1.5-3 g/ml;
and/or the concentration of the carboxymethyl cellulose in the system is 5-15g/ml, preferably 8-12 g/ml.
8. A microbial fertilizer prepared from the microbial agent prepared by the preparation method of any one of claims 1 to 7.
9. A method of preparing a microbial fertilizer as claimed in claim 8 comprising the steps of:
s11, mixing animal or poultry excrement with straws and leaves to prepare a composting raw material;
s12, controlling the water content of the composting raw material between 50 and 55 percent;
s13, mixing and fermenting the microbial inoculum and the compost raw material with the regulated moisture, and turning the compost during the mixing and fermentation to control the fermentation temperature;
and S14, decomposing the raw materials to obtain the microbial fertilizer.
10. The method according to claim 9, wherein the amount of the microbial agent to be added is determined in S13 by:
s131, using microbial fertilizer samples obtained with different addition amounts as arguments, U ═ x1, x2, …, Xn }, xi ═ (xi1, xi2, xi3), i ═ 1,2, …, n, using the color, odor, and fecal mass particle diameter of the microbial fertilizer samples as properties;
s132, standard deviation and range conversion is carried out through the following formulas:
Figure FDA0002904097860000021
Figure FDA0002904097860000022
is mean value, SkIs the standard deviation;
Figure FDA0002904097860000023
s133, order
Figure FDA0002904097860000024
Wherein, C is a constant and takes a value between 0.38 and 0.40;
s134, obtaining a transfer closure matrix through a cubic flat method according to
Figure FDA0002904097860000028
The size of (2), classifying;
s135, if
Figure FDA0002904097860000025
If the additive amount is at least divided into two types, determining the additive amount in the class with the minimum additive amount as the alternative additive amount;
s136, if
Figure FDA0002904097860000026
Repeating S131-S134 until
Figure FDA0002904097860000027
And S137, repeating the steps for 3 times, and selecting the minimum candidate addition amount as the addition amount.
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