CN108728379B - Composite microbial inoculum for quickly decomposing livestock and poultry manure and preparation method thereof - Google Patents

Composite microbial inoculum for quickly decomposing livestock and poultry manure and preparation method thereof Download PDF

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CN108728379B
CN108728379B CN201810557239.7A CN201810557239A CN108728379B CN 108728379 B CN108728379 B CN 108728379B CN 201810557239 A CN201810557239 A CN 201810557239A CN 108728379 B CN108728379 B CN 108728379B
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赵旭
王文丽
李娟�
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INSTITUTE OF SOIL FERTILIZER AND WATER-SAVING AGRICULTURE GANSU ACADEMY OF AGRICULTURAL SCIENCES
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Abstract

The invention discloses a composite microbial inoculum for fast decomposing livestock and poultry manure, which comprises a bacillus safensis strain and a bacillus licheniformis strain; and its preparing process are also disclosed. The invention has the following beneficial effects: two strains in the composite microbial inoculum can be well symbiotic, and antagonism does not exist between the strains; the preparation method of the microbial inoculum is simple and easy to operate, and the preparation cost is low; the method is mainly applied to the rapid decomposition of the livestock and poultry manure compost in low-temperature seasons, has the effects of enhancing the decomposition of organic matters in the livestock and poultry manure, reducing the nitrogen loss rate, eliminating odor, shortening the decomposition time and improving the quality of decomposed products, and has various physical and chemical indexes superior to those of the compost without adding microbial inoculum. Compared with a control group, the livestock manure added with the microbial inoculum has the heap temperature rising speed obviously higher than that of the control group without any microbial inoculum, the compost water content falling speed is higher than that of the control group, and H2The release amount of S gas is reduced, the nitrogen loss rate is reduced, and the organic carbon of the compost product is improved.

Description

Composite microbial inoculum for quickly decomposing livestock and poultry manure and preparation method thereof
Technical Field
The invention relates to the technical field of microbial preparations, in particular to a compound microbial inoculum for quickly decomposing livestock and poultry manure and a preparation method thereof.
Background
With the continuous development of livestock and poultry breeding industry in China, the quantity of livestock and poultry wastes generated every year is increased year by year, if the wastes are not treated properly, the environment can be seriously polluted, and further, the health of human bodies and the continuous development of the livestock and poultry breeding industry are seriously threatened. How to treat the livestock manure in a harmless and resourceful way to the maximum extent can achieve the aims of improving the environment and promoting the economic development, and has important practical significance on the protection of resources and environment in China and the sustainable development of economy. High temperature aerobic composting is the most common and feasible method for treating livestock and poultry manure. The compost treatment can reduce the pollution of livestock and poultry manure to the environment to the maximum extent, and compost products can be used as fertilizers required by the growth of crops, so that the growth of the crops is promoted, and the yield of the crops is increased.
Aerobic composted materials contain a large number of microorganisms that break down the organic matter in the composted material by producing a degrading enzyme. The compost material is added with the exogenous microbial inoculum, so that the number and the types of microorganisms in the compost material are increased, the decomposition of organic substances is accelerated, and the quality of compost products is improved. In the study of Liuchangli and other external microbial inoculum added compost for preparing compost, compared with the compost without the microbial inoculum, the compost with the microbial inoculum has the heap temperature of 63.0 ℃ and the high-temperature period of more than 55.0 ℃ of 7 days, and the composting time is shortened by 17 days compared with the compost without the microbial inoculum. The tomato seeds and tomato seedlings cultured by the compost product grow better than the observation group added with the microbial inoculum obviously than the control group not added with the microbial inoculum. Researches of researchers also show that the odor of excrement can be eliminated, the nitrogen loss rate can be reduced, and the contents of organic matters, total nitrogen, total phosphorus and total potassium in compost products can be improved by adding the exogenous microbial agent.
Cellulose is a main component of cow dung, crop straws and other agricultural organic solid wastes and is a main raw material in the cow dung composting process, the content of the cellulose is a main factor limited in composting, but most of lignocellulose is difficult to degrade, and the decomposition of the compost is closely related to the degradation of cellulose. Researches show that the cellulose degradation bacteria are inoculated in the composting process, so that the cellulose degradation can be promoted, the mineralization speed of organic substances is accelerated, the carbon-nitrogen ratio in the composting process is reduced, and the decomposition rate is increased.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a complex microbial inoculum for quickly decomposing livestock and poultry manure and a preparation method thereof.
In order to achieve the purpose, the technical scheme provided by the invention is as follows: a composite microbial inoculum for quickly decomposing livestock and poultry manure, which comprises bacillus safensis(Bacillus safensis)Strain and bacillus licheniformis（Bacillus licheniformis)A strain;
the Bacillus safensis strain has a preservation number of CGMCC NO.15809 and is named as Bacillus safensis GWX1-2a, the preservation time is 5 months and 24 days in 2018, the preservation organization is the China general microbiological culture Collection center and is No. 3 of Xilu No.1 of Beijing university facing Yang district, and the institute of microbiology of China academy of sciences;
the bacillus licheniformis strain has the preservation number of CGMCC NO.15810 and is named as bacillus licheniformis X10, the preservation time is 5 months and 24 days in 2018, the preservation organization is the China general microbiological culture preservation management center and the address is No. 3 of Beijing university Hokko 1 of sunward area, China academy of sciences microbiology institute.
Further, the composite microbial inoculum for the rapid decomposition of the livestock and poultry manure is prepared by mixing a bacillus fomentarius strain and a bacillus licheniformis strain according to the volume ratio of 1: 1.
The second purpose of the invention is to provide a preparation method of the complex microbial inoculum for fast decomposing the livestock and poultry manure, which comprises the following steps:
1) preparing a bacillus safensis fermentation liquid:
in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus safensis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 12-33 hours, and the bacillus safensis fermentation liquid is stored for later use after the fermentation is finished;
2) preparing a bacillus licheniformis fermentation liquid:
in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus licheniformis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 6-25 hours, and the bacillus licheniformis fermentation liquid is stored for later use after the fermentation is finished;
3) preparing a complex microbial inoculum:
respectively taking 100-800 parts of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps 1) and 2), and respectively filling the two into sterilized containers for sealed storage, so as to obtain the finished product of the composite microbial inoculum.
Further, in the preparation method of the composite microbial inoculum for quickly decomposing the livestock and poultry manure, in the step 1) and the step 2), the viable bacteria amount of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor is more than 2 multiplied by 109cfu/mL。
Further, in the step 1), the fermentation medium is prepared by adding 5.0g of peptone, 0.125g of NaCl and K into 1000ml of water2HPO4 0.5g，MgSO4 0.2g，FeSO40.15g, and adjusting the pH value to 7.2-7.4.
Further, in the step 2), the fermentation medium is prepared by adding 5.0g of CMC-Na, 5.0g of peptone, 0.125g of NaCl, and K into 1000ml of water2HPO4 0.5g，MgSO40.2g, and adjusting the pH value to 7.2-7.4.
Further, in the step 3), the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor are uniformly mixed according to the volume ratio of 1: 1.
The invention relates to a strain in a microbial agent for quickly decomposing livestock and poultry manure, which comprises the following components in percentage by weight: the bacillus safensis strain and the bacillus licheniformis strain are strains which are independently separated from the research institute of soil fertilizer and water conservation of agricultural academy of science of Gansu province and are identified as bacillus safensis and bacillus licheniformis by the research institute of microorganisms of the Chinese academy of science.
The invention provides a microbial agent for fast decomposing livestock and poultry manure, which is mainly characterized in that bacillus licheniformis and bacillus safensis are respectively subjected to liquid fermentation, so that the viable bacteria amount in fermentation liquor is more than 2 multiplied by 109cfu/mL; then mixing the two fermentation liquors according to the proportion of 1:1 in a sterile environment, finally subpackaging the mixture in a sterilized container for sealed storage, and preparing the high-efficiency microbial inoculum which is a liquid microbial inoculum and can be directly used.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention takes bacillus safensis and bacillus licheniformis as strains, and the bacillus safensis and the bacillus licheniformis are prepared by mixing the strains according to the ratio of 1:1 after fermentation in a fermentation tank, and the two strains can well symbiotic without antagonism between the strains.
2. The preparation method of the livestock and poultry manure efficient and rapid decomposition microbial inoculum provided by the invention is simple and easy to operate, and has low preparation cost.
3. The high-efficiency and quick-decomposing microbial inoculum for the livestock and poultry manure is mainly applied to quick decomposition of compost of the livestock and poultry manure (cow manure) in low-temperature seasons, has the effects of enhancing the decomposition of organic matters in the livestock and poultry manure, reducing the nitrogen loss rate, eliminating odor, shortening the decomposing time and improving the quality of decomposed products, and has various physical and chemical indexes superior to those of compost without microbial inoculum. Compared with a control group, the livestock manure added with the microbial inoculum has the heap temperature rising speed obviously higher than that of the control group without any microbial inoculum, the compost water content falling speed is higher than that of the control group, and H2The release amount of S gas is reduced by 54.24%, the nitrogen loss rate is reduced by 58.77%, and the organic carbon of the compost product is improved by 8.61%.
Drawings
FIG. 1 shows the treatment of shafudi bacterial agent (patent of invention) and no bacterial agent, and is commercially available
Figure DEST_PATH_IMAGE002
Microbial inoculum, commercially available
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Microbial inoculum, commercially available
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Bacterial agent cow dung compost H2And S release amount is compared.
FIG. 2 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 798153DEST_PATH_IMAGE002
Microbial inoculum, commercially available
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Microbial inoculum, commercially available
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NH of microbial inoculum cow dung compost3And (4) comparing the release amount.
FIG. 3 shows the sandford fungicide (patent of invention) and no fungicideTreatment of (2) and marketing
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Microbial inoculum, commercially available
Figure 16197DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 963293DEST_PATH_IMAGE006
And (4) comparing the germination indexes of the microbial inoculum cow dung compost.
FIG. 4 shows the treatment of shafudi bacterial agent (patent of invention) and no bacterial agent, and is commercially available
Figure 230326DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 429226DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 35788DEST_PATH_IMAGE006
Comparing the total nitrogen change of the microbial inoculum cow dung compost.
FIG. 5 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 29152DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 783481DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 707444DEST_PATH_IMAGE006
And (4) comparing the change of organic carbon of the microbial inoculum cow dung compost.
FIG. 6 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 230829DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 598357DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 839982DEST_PATH_IMAGE006
The quantity of fecal colibacillus of the microbial inoculum cow dung compost is compared.
FIG. 7 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 115106DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 945527DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 280694DEST_PATH_IMAGE006
And (3) comparing the pH value of the microbial inoculum cow dung compost.
FIG. 8 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 947298DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 26113DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 524090DEST_PATH_IMAGE006
The water content of the compost of the microbial inoculum cow dung is compared.
FIG. 9 shows the treatment of shafudi bacterial agent (patent of invention) and no addition of bacterial agent, and is commercially available
Figure 482688DEST_PATH_IMAGE002
Microbial inoculum, commercially available
Figure 433326DEST_PATH_IMAGE004
Microbial inoculum, commercially available
Figure 315832DEST_PATH_IMAGE006
The compost temperature change of the microbial inoculum cow dung compost is compared. Wherein, after the sandofan microbial inoculum (invention patent) is added, the pile temperature rises quickly in the initial composting period and the high temperature period lastsThe time is short.
Detailed Description
Example 1:
the formula of the microbial agent for quickly decomposing the livestock and poultry manure is as follows: 500 parts of bacillus licheniformis fermentation liquor and 500 parts of bacillus safensis fermentation liquor.
The preparation method comprises the following steps:
(1) preparing a bacillus safensis fermentation liquid:
the fermentation medium with pH of 7.2 was used for the cultivation (medium components: peptone 5.0g, NaCl 0.125g, K)2HPO4 0.5g，MgSO4 0.2g，FeSO40.15g of water, 1000mL of water), the inoculation amount of the seed liquid is 1.5 percent, the fermentation temperature is 37 ℃, the stirring speed is 300r/min, the optimal ventilation amount is 6.5L/min, and the fermentation is carried out for 12 hours to prepare the bacillus safensis fermentation liquid for later use;
(2) preparing a bacillus licheniformis fermentation liquid:
culturing with pH 7.0 fermentation medium (medium components: CMC-Na 5.0g, peptone 5.0g, NaCl 0.125g, and K)2HPO4 0.5g，MgSO40.2g of water, 1000mL of water), the seed liquid inoculation amount is 2%, the fermentation temperature is 35 ℃, the stirring speed is 320r/min, the optimal ventilation amount is 6.5L/min, the fermentation is carried out for 10 hours, and the fermentation liquid is stored for later use after the fermentation is finished.
(3) Preparation of the microbial inoculum:
and (3) taking 500 parts of each of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps (1) and (2), mixing and stirring uniformly, and filling the mixture into a sterilized container for sealing and storage, thus obtaining the finished microbial inoculum.
Example 2:
the formula of the microbial agent for quickly decomposing cow dung is as follows: 800 parts of bacillus licheniformis fermentation liquor and 800 parts of bacillus safensis fermentation liquor.
The preparation method comprises the following steps:
(1) preparing a bacillus safensis fermentation liquid:
the culture was carried out using a fermentation medium of pH 7.4 (medium composition: peptone 5.0g, N)aCl 0.125g，K2HPO4 0.5g，MgSO4 0.2g，FeSO40.15g of water, 1000mL of water), the inoculation amount of the seed liquid is 1.5 percent, the fermentation temperature is 35 ℃, the stirring speed is 280r/min, the optimal ventilation amount is 6L/min, and the fermentation is carried out for 24 hours to prepare the bacillus safensis fermentation liquid for later use;
(2) preparing a bacillus licheniformis fermentation liquid:
culturing with pH 7.2 fermentation medium (medium components: CMC-Na 5.0g, peptone 5.0g, NaCl 0.125g, and K)2HPO4 0.5g，MgSO40.2g of water, 1000mL of water), the seed liquid inoculation amount is 1%, the fermentation temperature is 33 ℃, the stirring speed is 320r/min, the optimal ventilation amount is 5.5L/min, the fermentation is carried out for 15 hours, and the fermentation liquid is stored for later use after the fermentation is finished.
(3) Preparation of the microbial inoculum:
and (3) taking 800 parts of each of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps (1) and (2), mixing and stirring uniformly, and filling the mixture into a sterilized container for sealing and storage, so that the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor can be finished microbial inoculum.
Example 3:
the formula of the microbial agent for quickly decomposing cow dung is as follows: 800 parts of bacillus licheniformis fermentation liquor and 200 parts of bacillus safensis fermentation liquor.
The preparation method comprises the following steps:
(1) preparing a bacillus safensis fermentation liquid:
the fermentation medium with pH of 7.3 was used for the cultivation (medium components: peptone 5.0g, NaCl 0.125g, K)2HPO4 0.5g，MgSO4 0.2g，FeSO40.15g of water, 1000mL of water), the inoculation amount of the seed liquid is 2%, the fermentation temperature is 37 ℃, the stirring speed is 300r/min, the optimal ventilation amount is 6.5L/min, and the fermentation is carried out for 30 hours to obtain the bacillus safensis fermentation liquid for later use;
(2) preparing a bacillus licheniformis fermentation liquid:
culturing with pH 7.1 fermentation medium (medium components: CMC-Na 5.0g, peptone 5.0g, NaCl 0.125g, and K)2HPO4 0.5g，MgSO40.2g of water, 1000mL of water), the seed liquid inoculation amount is 0.5%, the fermentation temperature is 37 ℃, the stirring speed is 300r/min, the optimal ventilation amount is 6.5L/min, the fermentation is carried out for 24 hours, and the fermentation liquid is stored for later use after the fermentation is finished.
(3) Preparation of the microbial inoculum:
and (3) taking 200 parts of each of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps (1) and (2), mixing and stirring uniformly, and filling the mixture into a sterilized container for sealing and storage, thus obtaining the finished microbial inoculum.
Example 4:
the composting effects of the microbial agents are compared, as shown in figures 1-9:
according to the compost contrast test of the microbial agent, the microbial agent for quickly decomposing the livestock and poultry manure is mainly adopted, the microbial agent is added according to the volume weight ratio of 1 per thousand, the microbial agent and the cow dung are uniformly mixed, the water content of the cow dung is adjusted to about 65 percent, and a pile with the width of 1.5 meters, the height of 1 meter and the length of 20 meters is built for compost fermentation. The main composting effects are as follows:
(1) the composting temperature of the cow dung added with the microbial inoculum is increased to 60.33 ℃ on 8 days, while the cow dung of a control group without any microbial inoculum is increased to 61.67 ℃ on 10 days, and the heap temperature increasing speed of the cow dung added with the microbial inoculum is obviously higher than that of the cow dung of the control group without any microbial inoculum.
(2) The water content of the compost of the cow dung added with the microbial agent is reduced faster than that of a control group.
(3) Compared with cow dung of a control group without any microbial inoculum, the cow dung added with the microbial inoculum of the invention has the advantages that H is higher2The release amount of S gas is reduced by 54.24 percent, and the odor of the cow dung compost is effectively reduced.
(4) Compared with cow dung of a control group without any microbial inoculum, the cow dung added with the microbial inoculum has the advantages that the organic carbon of a compost product is improved by 8.61%, and the nitrogen loss rate is reduced by 58.77%.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (5)

1. A composite microbial inoculum for quickly decomposing livestock and poultry manure is characterized by comprising bacillus safensis(Bacillus safensis)Strain and bacillus licheniformis(Bacillus licheniformis)Strain composition;
the Bacillus safensis strain has a preservation number of CGMCC NO.15809 and is named as Bacillus safensis GWX1-2a, the preservation time is 2018, 5 and 24 days, and the preservation organization is the China general microbiological culture preservation management center;
the bacillus licheniformis strain has a preservation number of CGMCC NO.15810 and is named as bacillus licheniformis X10, the preservation time is 5 months and 24 days in 2018, and the preservation organization is the China general microbiological culture preservation management center;
the preparation method of the complex microbial inoculum comprises the following steps:
1) preparing a bacillus safensis fermentation liquid: in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus safensis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 12-33 hours, and the bacillus safensis fermentation liquid is stored for later use after the fermentation is finished;
2) preparing a bacillus licheniformis fermentation liquid: in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus licheniformis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 6-25 hours, and the bacillus licheniformis fermentation liquid is stored for later use after the fermentation is finished;
3) preparing a complex microbial inoculum: respectively taking 100-800 parts of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps 1) and 2), and respectively filling the two into sterilized containers for sealed storage, so as to obtain a finished product of the composite microbial inoculum;
in the step 1), the proportion of the fermentation medium is that peptone 5.0g, NaCl 0.125g and K are added into every 1000ml of water2HPO4 0.5g，MgSO4 0.2g，FeSO40.15g, adjusting the pH value to 7.2-7.4;
in the step 2), the proportion of the fermentation medium is that every 1000ml of water is added with 5.0g of CMC-Na, 5.0g of peptone, 0.125g of NaCl and K2HPO4 0.5g，MgSO40.2g, and adjusting the pH value to 7.2-7.4.
2. The complex microbial inoculant for the rapid decomposition of livestock and poultry manure as claimed in claim 1, wherein the complex microbial inoculant is prepared by mixing a bacillus saffron strain and a bacillus licheniformis strain according to a volume ratio of 1: 1.
3. The preparation method of the complex microbial inoculum for the rapid decomposition of the livestock and poultry manure as claimed in claim 1, which is characterized by comprising the following steps:
1) preparing a bacillus safensis fermentation liquid: in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus safensis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 12-33 hours, and the bacillus safensis fermentation liquid is stored for later use after the fermentation is finished;
2) preparing a bacillus licheniformis fermentation liquid: in a fermentation tank, the liquid loading amount of a fermentation medium is 55-65%, the inoculation amount of a bacillus licheniformis seed liquid is 0.5-2%, the fermentation temperature is 33-37 ℃, the stirring speed is 280-320 r/min, the ventilation amount is 5.5-6.5L/min, the fermentation end point is 6-25 hours, and the bacillus licheniformis fermentation liquid is stored for later use after the fermentation is finished;
3) preparing a complex microbial inoculum: respectively taking 100-800 parts of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor prepared in the steps 1) and 2), and respectively filling the two into sterilized containers for sealed storage, so as to obtain the finished product of the composite microbial inoculum.
4. The method for preparing the composite microbial inoculum for the rapid decomposition of the livestock and poultry manure according to the claim 3, wherein in the step 1) and the step 2), the viable bacteria amount of the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor is more than 2 x109cfu/mL。
5. The method for preparing the complex microbial inoculum for the rapid decomposition of the livestock and poultry manure according to the claim 3, wherein in the step 3), the bacillus safensis fermentation liquor and the bacillus licheniformis fermentation liquor are uniformly mixed according to the volume ratio of 1: 1.
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