CN118006486B - Composite microbial flora for sheep manure composting - Google Patents

Composite microbial flora for sheep manure composting Download PDF

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CN118006486B
CN118006486B CN202410105453.4A CN202410105453A CN118006486B CN 118006486 B CN118006486 B CN 118006486B CN 202410105453 A CN202410105453 A CN 202410105453A CN 118006486 B CN118006486 B CN 118006486B
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CN118006486A (en
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李馥霞
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Linyi Academy of Agricultural Sciences
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Abstract

The invention discloses a composite microbial flora for sheep manure composting, and belongs to the technical field of microorganisms. The flora comprises functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10. The microbial flora for fermenting the sheep manure is prepared by compounding functional microorganisms, auxiliary microorganisms, fermentation promoters, microbial carriers and the like, and can promote rapid temperature rise of compost, perform efficient fermentation, effectively degrade cellulose substances in the sheep manure, promote the increase of organic matters and total nutrient content in fermentation products, effectively complete harmless treatment of the sheep manure, and have remarkable economic and social benefits.

Description

Composite microbial flora for sheep manure composting
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a composite microbial flora for sheep manure composting.
Background
With the rapid development of economy, the living standard of people is continuously improved, the consumption demand of people for livestock products is also increased year by year, the large-scale and intensive degree of livestock and poultry breeding production is continuously improved, and the problem of livestock and poultry manure treatment is more remarkable. At present, livestock and poultry manure in livestock and poultry farms becomes an important source of agricultural non-point source pollution. The livestock and poultry manure contains rich organic matters, nitrogen, phosphorus, potassium and other microelements, and can be a huge resource library if being reasonably utilized. However, the phenomenon that the livestock and poultry manure is inappropriately utilized in a resource manner is still common, and serious environmental pollution and resource waste are caused.
The main modes of fertilizer, feed and energy are the main modes of livestock and poultry manure resource utilization, and the most common treatment mode is composting. Composting is one of the means for treating various kinds of animal manure in China with long history. The method is a biological process of decomposing organic matters in the livestock and poultry feces into stabilized and harmless humus by means of high-temperature fermentation under the condition of manually controlling fermentation temperature, moisture and ventilation and by means of microorganisms such as bacteria and fungi contained in the feces or manually adding a microbial agent. In the composting process, unstable organic matters are converted into stable nutrients which can be directly absorbed and utilized by plant roots by microorganisms, pathogenic bacteria, ova and weed seeds in the feces are killed by the high temperature generated in the fermentation process, so that the breeding of weeds and pests can be reduced when the organic matters are applied to the field, and the organic fertilizer is not only an excellent soil conditioner, but also a plump organic fertilizer. Meanwhile, the bio-compost fermentation has the characteristics of low operation cost, small secondary pollution and the like, and is one of the ideal and effective modes for harmless treatment of the excrement.
Compared with other animal manure, the sheep manure has higher organic matters, nitrogen, phosphorus and potassium content and richer nutrients, and is a good organic fertilizer resource. However, sheep are typical herbivores, and the manure contains a large amount of cellulose substances which are difficult to decompose, so that composting effect is severely restricted, composting fermentation time is long, organic matter conversion efficiency is low, and the like.
Microbiota plays an important role in the composting process. A great deal of research shows that inoculating exogenous microorganisms is an effective way to accelerate the high-temperature aerobic composting process. The addition of specific types of microorganisms can accelerate the decomposition of the initial matrixes of the compost, accelerate the degradation of lignocellulose, promote the post-maturation of the piled bodies and shorten the composting period. Although various microbial composting corrosion promoting microbial agents exist in the domestic market at present, the microbial composting corrosion promoting microbial agents are often used for sheep manure composting fermentation, and the problems of long fermentation period, low decomposition degree, incomplete degradation of cellulose substances and the like exist. Therefore, proper strains are required to be screened according to the types of fermentation materials so as to ensure the fermentation effect.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a composite microbial agent for efficiently fermenting sheep manure, which comprises functional microorganisms, auxiliary microorganisms, fermentation promoters, microorganism carriers and other components, can efficiently decompose the sheep manure, and can realize rapid and high-degree degradation of cellulose in the sheep manure.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
A composite microbial flora for sheep manure composting comprises functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10.
Further, the functional microorganisms are bacillus megatherium (Bacillus megaterium), lactobacillus plantarum (Lactobacillus plantarum) and marine bacillus aceosus (Pontibacter akesuensis); the preservation number of the bacillus megatherium is CGMCC No.1.16094, the bacillus megatherium is purchased from China general microbiological culture Collection center, and the original preservation time is 2017, 3, 11 days; the lactobacillus plantarum has a preservation number of CGMCC No.1.12935, is purchased from China general microbiological culture Collection center, and has an original preservation time of 2014, 6 months and 7 days; the collection number of the oceanic Acer kesu is CGMCC No.1.12365, the oceanic Acer kesu is purchased from China general microbiological collection center, and the original collection date is 10, 15 and 2012. The three functional microorganisms can be inquired and purchased through the strain catalogue of the China general microbiological deposit management center, and repeated biological deposit is not needed.
Further, the mass ratio of the bacillus megatherium to the lactobacillus plantarum to the marine bacillus aceosum is 1:1:1.
The preparation method of the functional microorganism comprises the following steps: activating bacillus megatherium, lactobacillus plantarum and marine Acetes in LB solid culture medium respectively, then picking single colony, inoculating the single colony into test tubes of LB liquid culture medium respectively, and culturing at 30-37 ℃ at 180-200r/min until OD 600 = 1.2 to obtain seed liquid; inoculating the seed solution into 100mLLB liquid culture medium according to 2% quantity, culturing for 12h at 30-37 ℃ and 180-200r/min, washing 3 times with sterile PBS buffer solution, then resuspending to obtain three bacterial suspensions, respectively spray-drying, and mixing the three bacterial powders according to the mass ratio of 1:1:1 to obtain the functional microorganism.
Furthermore, the number of effective viable bacteria in the three bacterial suspensions is not less than 1X 10 9 CFU/mL.
More preferably, the preparation method of the LB liquid medium comprises the following steps: mixing 1L of water, 12g of tryptone, 6g of yeast extract and 11g of NaCl uniformly until solute is dissolved, and regulating the pH to 7.0 by using 1-3mol/L of NaOH to obtain an LB liquid culture medium; the LB solid culture medium is obtained by adding 10-15g agar based on LB liquid culture medium.
Further, the auxiliary microorganism comprises bacillus subtilis, bacillus licheniformis, saccharomycetes, trichoderma viride and aspergillus niger, and the mass ratio is 1:1: (1-3): (0.5-1): (0.5-1).
The auxiliary microorganisms such as bacillus subtilis, bacillus licheniformis, saccharomycetes and trichoderma viride are used for purchasing the commercial common microbial agents.
Further, the microbial fermentation promoter is potassium dihydrogen phosphate, urea and sucrose according to the following proportion (0.05-0.1): (0.05-0.1): (0.1-0.5) by weight ratio.
Further, the microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of alkali carbon of 2:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
Further, the solvent in the step (2) is a mixed solution of ethanol and water, wherein the volume ratio of the ethanol to the water is 1:1.
The preparation method of the flora comprises the following steps: uniformly mixing functional microorganisms, auxiliary microorganisms, a microorganism fermentation promoter and a microorganism carrier according to the mass ratio of 1:0.5:0.3:10, and according to the solid-liquid ratio of 1g: dispersing 10mL in water, stirring thoroughly, soaking for 10-12 hr to allow the carrier to adsorb microorganism thoroughly, drying, packaging, and warehousing.
The microbial inoculum of the invention has the use amount of 0.3 percent (mass ratio) of the raw material of the compost sheep manure.
Sheep manure curing is a process with continuously changing temperature, and when the composting temperature is lower than or higher than the optimal temperature of microbial flora, the microbial enzyme production efficiency is low or even no enzyme is produced, so that the degradation efficiency of cellulose in the livestock manure composting process is low, and the time is long. The invention screens three microorganism strains for producing cellulase in high yield to form functional microorganisms, the optimal enzyme production temperature of bacillus megatherium, lactobacillus plantarum and marine bacillus aceosum corresponds to the temperature of different stages of high, medium and low composting fermentation, and the enzyme can be continuously produced at different fermentation temperature stages in the composting process after being mixed in equal proportion, thus promoting the decomposition of cellulose in livestock manure under synergistic effect. The synergistic effect of the three functional microorganisms can rapidly improve the level and peak value of cellulase activity of livestock and poultry manure fermentation compost, shorten the fermentation time of the compost, and have good adaptability and decomposition effect on the compost.
Meanwhile, the auxiliary microorganisms such as the bacillus subtilis, the bacillus licheniformis, the saccharomycetes, the trichoderma viride and the aspergillus niger are added, so that the bacillus subtilis can obviously reduce mineralization of compost and mature stages and promote humification of carbon, thereby reducing carbon dioxide emission and obviously improving humic acid content in the compost and quality of a compost product. The bacillus licheniformis, saccharomycetes and other strains can efficiently degrade easily degradable organic components in the compost materials in the initial fermentation period, so that the temperature rise of the compost is promoted rapidly. And the trichoderma viride, aspergillus niger and the like can play a role in promoting fermentation, and the stable and effective fermentation process is kept through secreting various active enzymes.
The invention adds a little potassium dihydrogen phosphate, urea and sucrose to form a microbial accelerant, thereby providing supplementary nutrition for microbial activities. Simultaneously preparing biomass carbon by adopting coconut shells as a microbial carrier, carrying out pyrolysis on the coconut shells under the anoxic condition, forming a honeycomb dense porous structure by virtue of the etching action of a proper amount of activator sodium hydroxide, adsorbing and fixing microorganisms, and enabling a large amount of aromatic substances and functional groups contained in the biomass carbon to simultaneously adsorb anions and cations, and also forming firm covalent bonds with the functional groups on the surfaces of the microorganisms to fix the microorganisms, so that the biomass carbon can also adsorb and stabilize the microorganisms; meanwhile, the biochar has an effect of improving the pore structure of the pile body, can promote composting and decomposition, reduce greenhouse gas emission in the composting process, accelerate mineralization and humification of organic matters, and increase nutrient content of compost products.
Advantageous effects
The microbial flora for fermenting the sheep manure is prepared by compounding functional microorganisms, auxiliary microorganisms, fermentation promoters, microbial carriers and the like, and can promote rapid temperature rise of compost, perform efficient fermentation, effectively degrade cellulose substances in the sheep manure, promote the increase of organic matters and total nutrient content in fermentation products, effectively complete harmless treatment of the sheep manure, and have remarkable economic and social benefits.
Drawings
FIG. 1 is a graph showing the change of cellulase activity with time in the fermentation of sheep manure by applying the flora of example 3 of the present invention;
FIG. 2 is an electron microscopic image of the internal morphology of the porous biochar of the microbial carrier of example 3 and comparative examples 10-11 of the present invention.
Detailed Description
The technical scheme of the present invention is further described below with reference to specific examples, but is not limited thereto.
Example 1
A composite microbial flora for sheep manure composting comprises functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10.
The functional microorganisms are bacillus megatherium, lactobacillus plantarum and marine Acetes; the preservation number of the bacillus megatherium is CGMCC No.1.16094, the bacillus megatherium is purchased from China general microbiological culture Collection center, and the original preservation time is 2017, 3, 11 days; the lactobacillus plantarum has a preservation number of CGMCC No.1.12935, is purchased from China general microbiological culture Collection center, and has an original preservation time of 2014, 6 months and 7 days; the collection number of the oceanic Acer kesu is CGMCC No.1.12365, the oceanic Acer kesu is purchased from China general microbiological collection center, and the original collection date is 10, 15 and 2012. All three functional microorganisms can be inquired and purchased through the strain catalogue of China general microbiological deposit management center without repeated biological deposit.
The mass ratio of the bacillus megatherium to the lactobacillus plantarum to the marine bacillus acer is 1:1:1.
The preparation method of the functional microorganism of the embodiment comprises the following steps: activating bacillus megatherium, lactobacillus plantarum and marine Acetes in LB solid culture medium respectively, then picking single colony, inoculating the single colony into test tubes of LB liquid culture medium respectively, and culturing at 30-37 ℃ at 180-200r/min until OD 600 = 1.2 to obtain seed liquid; inoculating the seed solution into 100mLLB liquid culture medium according to 2% quantity, culturing for 12h at 30-37 ℃ and 180-200r/min, washing 3 times with sterile PBS buffer solution, then resuspending to obtain three bacterial suspensions, respectively spray-drying, and mixing the three bacterial powders according to the mass ratio of 1:1:1 to obtain the functional microorganism.
The number of effective viable bacteria in the three bacterial suspensions is not less than 1X 10 9 CFU/mL.
The preparation method of the LB liquid medium comprises the following steps: mixing 1L of water, 12g of tryptone, 6g of yeast extract and 11g of NaCl uniformly until solute is dissolved, and regulating the pH value to 7.0 by using 1mol/L NaOH to obtain an LB liquid culture medium; the LB solid medium is obtained by adding 10g of agar based on LB liquid medium.
The auxiliary microorganism comprises bacillus subtilis, bacillus licheniformis, saccharomycetes, trichoderma viride and aspergillus niger, and the mass ratio is 1:1:1:1:0.5.
The auxiliary microorganisms such as bacillus subtilis, bacillus licheniformis, saccharomycetes and trichoderma viride are used for purchasing the commercial common microbial agents.
The microbial fermentation promoter is prepared from potassium dihydrogen phosphate, urea and sucrose according to the following proportion (0.05): (0.05): (0.5) by weight ratio.
The microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of alkali carbon of 2:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
The solvent in the step (2) is a mixed solution of ethanol and water, wherein the volume ratio of the ethanol to the water is 1:1.
The preparation method of the flora in the embodiment comprises the following steps: uniformly mixing functional microorganisms, auxiliary microorganisms, a microorganism fermentation promoter and a microorganism carrier according to the mass ratio of 1:0.5:0.3:10, and according to the solid-liquid ratio of 1g: dispersing 10mL in water, stirring thoroughly, soaking for 10 hr to allow the carrier to adsorb microorganism thoroughly, drying, packaging, and warehousing.
Example 2
A composite microbial flora for sheep manure composting comprises functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10.
The functional microorganisms are bacillus megatherium, lactobacillus plantarum and marine Acetes; the preservation number of the bacillus megatherium is CGMCC No.1.16094, the bacillus megatherium is purchased from China general microbiological culture Collection center, and the original preservation time is 2017, 3, 11 days; the lactobacillus plantarum has a preservation number of CGMCC No.1.12935, is purchased from China general microbiological culture Collection center, and has an original preservation time of 2014, 6 months and 7 days; the collection number of the oceanic Acer kesu is CGMCC No.1.12365, the oceanic Acer kesu is purchased from China general microbiological collection center, and the original collection date is 10, 15 and 2012. All three functional microorganisms in the embodiment can be inquired and purchased through the strain catalogue of the China general microbiological deposit management center without repeated biological deposit.
The mass ratio of the bacillus megatherium to the lactobacillus plantarum to the marine bacillus acer is 1:1:1.
The preparation method of the functional microorganism of the embodiment comprises the following steps: activating bacillus megatherium, lactobacillus plantarum and marine Acetes in LB solid culture medium respectively, then picking single colony, inoculating the single colony into test tubes of LB liquid culture medium respectively, and culturing at 30-37 ℃ at 180-200r/min until OD 600 = 1.2 to obtain seed liquid; inoculating the seed solution into 100mLLB liquid culture medium according to 2% quantity, culturing for 12h at 30-37 ℃ and 180-200r/min, washing 3 times with sterile PBS buffer solution, then resuspending to obtain three bacterial suspensions, respectively spray-drying, and mixing the three bacterial powders according to the mass ratio of 1:1:1 to obtain the functional microorganism.
The number of effective viable bacteria in the three bacterial suspensions is not less than 1X 10 9 CFU/mL.
The preparation method of the LB liquid medium comprises the following steps: mixing 1L of water, 12g of tryptone, 6g of yeast extract and 11g of NaCl uniformly until solute is dissolved, and regulating the pH to 7.0 by 2mol/L NaOH to obtain an LB liquid culture medium; the LB solid medium is obtained by adding 13g of agar based on LB liquid medium.
The auxiliary microorganism comprises bacillus subtilis, bacillus licheniformis, saccharomycetes, trichoderma viride and aspergillus niger, and the mass ratio is 1:1:2:0.5:1.
The auxiliary microorganisms such as bacillus subtilis, bacillus licheniformis, saccharomycetes and trichoderma viride are used for purchasing the commercial common microbial agents.
The microbial fermentation promoter is prepared from potassium dihydrogen phosphate, urea and sucrose according to the following proportion (0.1): (0.1): (0.1) by weight ratio.
The microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of alkali carbon of 2:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
The solvent in the step (2) is a mixed solution of ethanol and water, wherein the volume ratio of the ethanol to the water is 1:1.
The preparation method of the flora in the embodiment comprises the following steps: uniformly mixing functional microorganisms, auxiliary microorganisms, a microorganism fermentation promoter and a microorganism carrier according to the mass ratio of 1:0.5:0.3:10, and according to the solid-liquid ratio of 1g: dispersing 10mL in water, stirring thoroughly, soaking for 10 hr to allow the carrier to adsorb microorganism thoroughly, drying, packaging, and warehousing.
Example 3
A composite microbial flora for sheep manure composting comprises functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10.
The functional microorganisms are bacillus megatherium, lactobacillus plantarum and marine Acetes; the preservation number of the bacillus megatherium is CGMCC No.1.16094, the bacillus megatherium is purchased from China general microbiological culture Collection center, and the original preservation time is 2017, 3, 11 days; the lactobacillus plantarum has a preservation number of CGMCC No.1.12935, is purchased from China general microbiological culture Collection center, and has an original preservation time of 2014, 6 months and 7 days; the collection number of the oceanic Acer kesu is CGMCC No.1.12365, the oceanic Acer kesu is purchased from China general microbiological collection center, and the original collection date is 10, 15 and 2012. All three functional microorganisms in the embodiment can be inquired and purchased through the strain catalogue of the China general microbiological deposit management center without repeated biological deposit.
The mass ratio of the bacillus megatherium to the lactobacillus plantarum to the marine bacillus acer is 1:1:1.
The preparation method of the functional microorganism of the embodiment comprises the following steps: activating bacillus megatherium, lactobacillus plantarum and marine Acetes in LB solid culture medium respectively, then picking single colony, inoculating the single colony into test tubes of LB liquid culture medium respectively, and culturing at 30-37 ℃ at 180-200r/min until OD 600 = 1.2 to obtain seed liquid; inoculating the seed solution into 100mLLB liquid culture medium according to 2% quantity, culturing for 12h at 30-37 ℃ and 180-200r/min, washing 3 times with sterile PBS buffer solution, then resuspending to obtain three bacterial suspensions, respectively spray-drying, and mixing the three bacterial powders according to the mass ratio of 1:1:1 to obtain the functional microorganism.
The number of effective viable bacteria in the three bacterial suspensions is not less than 1X 10 9 CFU/mL.
The preparation method of the LB liquid medium comprises the following steps: mixing 1L of water, 12g of tryptone, 6g of yeast extract and 11g of NaCl uniformly until solute is dissolved, and regulating the pH to 7.0 by using 3mol/L NaOH to obtain an LB liquid culture medium; the LB solid culture medium is obtained by adding 15g of agar on the basis of LB liquid culture medium.
The auxiliary microorganism comprises bacillus subtilis, bacillus licheniformis, saccharomycetes, trichoderma viride and aspergillus niger, and the mass ratio is 1:1:3:0.8:0.8.
The auxiliary microorganisms such as bacillus subtilis, bacillus licheniformis, saccharomycetes and trichoderma viride are used for purchasing the commercial common microbial agents.
The microbial fermentation promoter is prepared from potassium dihydrogen phosphate, urea and sucrose according to the following proportion (0.08): (0.08): (0.3) by weight ratio.
The microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of alkali carbon of 2:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
The solvent in the step (2) is a mixed solution of ethanol and water, wherein the volume ratio of the ethanol to the water is 1:1.
The preparation method of the flora in the embodiment comprises the following steps: uniformly mixing functional microorganisms, auxiliary microorganisms, a microorganism fermentation promoter and a microorganism carrier according to the mass ratio of 1:0.5:0.3:10, and according to the solid-liquid ratio of 1g:10mL of the solution is dispersed in water, fully and uniformly stirred, soaked for 12 hours, so that the carrier fully adsorbs microorganisms, and then dried, packaged and put in storage.
Comparative examples 1 to 9
The composition ratio of the functional microorganisms, that is, the mass ratio of Bacillus megaterium, lactobacillus plantarum and Marine Acinetobacter was changed, and a comparative example was set. The rest of the raw materials and the process are the same as in example 3. Mass ratio of bacillus megaterium, lactobacillus plantarum and marine alchohol in comparative example is shown in table 1:
TABLE 1 comparative example strain mass ratio
Bacillus megaterium Lactobacillus plantarum Marine bacillus of Ackersu
Comparative example 1 2 1 1
Comparative example 2 1 2 1
Comparative example 3 1 1 2
Comparative example 4 0 1 1
Comparative example 5 1 0 1
Comparative example 6 1 1 0
Comparative example 7 0 0 1
Comparative example 8 0 1 0
Comparative example 9 1 0 0
Comparative example 10
The preparation method of the microbial carrier porous biochar is changed, namely the dosage ratio of the activator sodium hydroxide to the precursor is changed, and the rest raw materials and the preparation method are the same as in example 3.
The microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of 1:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
Comparative example 11
The preparation method of the microbial carrier porous biochar is changed, namely the dosage ratio of the activating agent to the precursor is changed, and the rest raw materials and the preparation method are the same as in example 3.
The microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of 3:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
Performance testing
The composting raw material is fresh sheep manure, the microbial inoculum obtained in the examples and the comparative examples is used as a fermenting agent, the proportion of the fermenting agent to the composting raw material is 0.3 percent (mass ratio), the fermenting agent and the sheep manure are mixed and stirred uniformly, the water content of the material is regulated to be 55+/-5 percent, and the control standard is' the condition that a mass of material is tightly held by hands without liquid flowing out and the material is not scattered after loosening hands; the pile body is conical, the pile height is 1.2-1.4m, and the diameter is 2-3m. The composting period is 25d (2022, 5, 15, 6, 15 days), and the composting turning adopts mechanical turning and natural ventilation.
Turning over the compost for 1 time when the temperature of the compost reaches 50 ℃ in the composting fermentation process; the temperature of the pile is continuously increased, and when the temperature reaches about 60 ℃, the pile is turned over for 1 time; the compost continues to ferment, the high temperature (50-65 ℃) is maintained for about 10d, the temperature of the pile body is not higher than 70 ℃ in the high temperature fermentation process, and when the temperature of the pile body reaches about 65 ℃, the pile body needs to be turned over, cooled and aerated; and (3) entering a post-decomposition period, turning the stack for 1 time at intervals of 3-4d, and gradually reducing the temperature of the stack until the temperature approaches the ambient temperature.
Detection method
The sample for completing the sampling of the compost is a multipoint mixed sample. The organic content of each mixed sample was determined by the K 2Cr2O7 capacity method. The indexes such as C/N content, fecal coliform number, roundworm mortality and the like in each mixed sample are determined by adopting national agricultural standard NY525-2011, and the cellulase activity in each mixed sample stage is determined by adopting a 3.5-dinitrosalicylic acid chromogenic method.
Extracting fresh compost samples with deionized water for 3 hours at a solid-to-liquid ratio of 1:10 (mass/volume), and filtering to obtain compost extract. Respectively measuring the pH value and EC of the diffuse compost extracting solution by a pH meter and a conductivity meter; measuring absorbance values (marked as E4 and E6) of the compost leaching solution at 465 and 665nm wavelengths respectively by using an ultraviolet spectrophotometer, and calculating an E4/E6 value; 20mL of the leaching solution is taken in a 9cm culture dish, filter paper is filled, 20 Chinese cabbage seeds are added into the leaching solution, the germination rate and root length are measured after the culture is carried out for 48 hours under the dark condition of 20 ℃, and the Germination Index (GI) of the seeds is calculated according to the following formula:
Table 2 sample test results
From the experimental data, the samples of the compost in the embodiment of the invention have the organic matter content not lower than 60%, the carbon-nitrogen ratio lower than 20 and the GI value higher than 100% and reach a complete decomposition state. E4/E6 is one of the humification parameters for evaluating the degree of compost maturity, and the smaller the value is, the greater the degree of compost humification is. Compared with the examples, the comparative examples change the composition and the dosage of the functional microbial agent, the synergistic balance among the three strains is broken, the fermentation effect is reduced, and the humification degree and various nutrition indexes are weakened to different degrees. The continuous and effective secretion of the cellulase with high activity by the three microorganisms in the whole fermentation process is a key for realizing the rapid sheep manure fermentation, as shown in figure 1, the temperature of the compost is continuously increased along with the change of time in the early stage of the composting, the activity of the cellulase in the compost is increased until the fermentation is finished, and the activity of the cellulase is kept at a higher level of 2-2.1 U.g -1.
Meanwhile, in the comparative examples 10-11 in which the proportion of the activator is changed, the carrier shape is changed, the alkali-carbon ratio is too low, the etching reaming effect of sodium hydroxide is weaker, and a developed porous structure cannot be formed; and the alkali carbon ratio is too high, the etching effect of sodium hydroxide is too strong, partial pore structures are broken, and the excellent porous biochar material cannot be formed, as shown in figure 2. Thus, the proper alkali-carbon ratio helps to form a good microporous biochar, which is more advantageous to microbial adsorption and continuous function.
It should be noted that the above-mentioned embodiments are merely some, but not all embodiments of the preferred mode of carrying out the invention. It is evident that all other embodiments obtained by a person skilled in the art without making any inventive effort, based on the above-described embodiments of the invention, shall fall within the scope of protection of the invention.

Claims (5)

1. The composite microbial flora for sheep manure composting is characterized by comprising functional microorganisms, auxiliary microorganisms, a microbial fermentation promoter and a microbial carrier, wherein the mass ratio of the functional microorganisms to the auxiliary microorganisms to the microbial fermentation promoter to the microbial carrier is 1:0.5:0.3:10; the functional microorganisms are bacillus megatherium (Bacillus megaterium), lactobacillus plantarum (Lactobacillus plantarum) and marine Acetes (Pontibacter akesuensis); the preservation number of the bacillus megatherium is CGMCC No.1.16094, and the bacillus megatherium is purchased from China general microbiological culture Collection center; the lactobacillus plantarum has a preservation number of CGMCC No.1.12935 and is purchased from China general microbiological culture Collection center; the collection number of the oceanic Acetes is CGMCC No.1.12365, and the oceanic Acetes is purchased from China general microbiological collection center; the mass ratio of the bacillus megatherium to the lactobacillus plantarum to the marine bacillus acer is 1:1:1.
2. The composite microbial flora for sheep manure composting of claim 1, wherein the auxiliary microorganisms comprise bacillus subtilis, bacillus licheniformis, saccharomycetes, trichoderma viride and aspergillus niger in a mass ratio of 1:1: (1-3): (0.5-1): (0.5-1).
3. The composite microbial flora for sheep manure composting of claim 1, wherein the microbial fermentation promoter is prepared by mixing potassium dihydrogen phosphate, urea and sucrose according to the weight ratio of (0.05-0.1) to (0.05-0.5).
4. The composite microbial flora for sheep manure composting of claim 1, wherein the microbial carrier is porous biochar, and the preparation method comprises the following steps:
(1) Pulverizing and grinding coconut shell, drying, placing in a crucible, placing in a box-type atmosphere furnace, and heating from room temperature to 400-500 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, and maintaining for 1h after reaching a target temperature to obtain a precursor of activated carbon;
(2) Mixing sodium hydroxide and active carbon precursor according to the mass ratio of alkali carbon of 2:1, and then according to the solid-liquid ratio of 1g: adding 20mL of solvent, mixing, magnetically stirring for 30min, standing for 12h after stirring uniformly, placing in an oven at 80-100 ℃ for drying, placing the dried sample in a box-type furnace, heating from room temperature to 800 ℃ at a heating rate of 10 ℃/min in a nitrogen atmosphere with a flow rate of 500mL/min, preserving heat and activating for 2h, washing the sample to be neutral by hydrochloric acid and deionized water after activation is finished, and drying to obtain the porous activated carbon.
5. The composite microbial flora for sheep manure composting of claim 4, wherein the solvent in the step (2) is a mixed solution of ethanol and water, wherein the volume ratio of ethanol to water is 1:1.
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