CN114456985B - Microbial strain for high-temperature aerobic composting of biogas residues and application of microbial strain - Google Patents
Microbial strain for high-temperature aerobic composting of biogas residues and application of microbial strain Download PDFInfo
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Abstract
The invention relates to the technical field of environmental microorganisms, in particular to a microbial strain for high-temperature aerobic composting of biogas residues and application thereof, wherein the microbial strain for high-temperature aerobic composting of the biogas residues is Geobacillus lituus G6-1, and the preservation number is CCTCC NO: m2022022027. The Geobacillus lituunicus G6-1 is high-temperature resistant and salt resistant, can improve the temperature of a compost body, prolong the high-temperature period of compost, quickly decompose organic matters, accelerate the composting process of biogas residues, shorten the composting time, improve the material maturity and the quality of organic fertilizer products, and has high application value.
Description
Technical Field
The invention relates to the technical field of environmental microorganisms, in particular to a microbial strain for high-temperature aerobic composting of biogas residues and application thereof.
Background
With the acceleration of the urbanization process, the yield of municipal domestic waste is increasing day by day. More than 50% of the domestic garbage is wet garbage, and the yield of the wet garbage is increased rapidly after the garbage is classified. Anaerobic digestion is an effective treatment method for wet garbage, and can convert the wet garbage into renewable energy, namely methane, which also conforms to the development strategy of low carbon. However, a large amount of solid residue, namely biogas residues, can be generated during anaerobic digestion, and about 25 kg of biogas residues are generated when one ton of wet garbage is treated. The biogas residue mainly comprises cellulose which is difficult to degrade and a large amount of anaerobic microorganisms. The conventional biogas residue treatment mode is to dehydrate, dry and incinerate the biogas residue, but the treatment mode has high energy consumption and is not suitable for the development target of low carbon and the resource utilization of organic solid wastes.
The wet waste biogas residues contain a large amount of organic matters, humic acid, crude protein, amino acid and other nutritional ingredients, and have high recycling value. However, untreated biogas residues are potentially dangerous to soil microorganisms, crops and even humans. According to the comprehensive treatment scheme of 'harmlessness, reduction and recycling' of organic wastes, aerobic composting is a harmless and recycling efficient treatment mode. Aerobic composting is a process of converting organic matters in organic wastes into humus and releasing energy under the synergistic action of various microorganisms under the aerobic condition, and has positive significance for promoting the virtuous cycle of the organic wastes in agricultural production.
Although the biogas residues contain a large amount of endogenous microorganisms, the microorganisms are mainly anaerobic microorganisms in the process of anaerobic biogas production, are not suitable for the environmental conditions of subsequent high-temperature aerobic composting, have small effect on the aerobic composting, and only rely on the endogenous microorganisms in the biogas residues to carry out the aerobic composting, so that the organic matter conversion and decomposition effects are not ideal. Therefore, microorganisms capable of accelerating the decomposition and rapid decomposition of organic matters under the high-temperature aerobic condition are screened, a microbial agent suitable for high-temperature aerobic composting of biogas residues is developed, the composting time is shortened, and the method has great significance for resource utilization of the biogas residues.
Disclosure of Invention
In view of the above disadvantages of the prior art, the present invention provides a microbial strain for high temperature aerobic composting of biogas residues and an application thereof, which are used for solving the problems in the prior art.
In order to achieve the aim and other related objects, the invention provides Geobacillus lituunicus G6-1 of Geobacillus lituus with the preservation number of CCTCC NO: M2022022027.
The invention also provides a liquid microbial inoculum, which comprises the Geobacillus lituus G6-1, wherein the concentration of the Geobacillus lituus G6-1 in the liquid microbial inoculum is up toAt a minimum 1X 10 8 CFU/mL。
The invention also provides a preparation method of the liquid microbial inoculum, which comprises the following steps: inoculating the pure strain of Geobacillus lituranicus G6-1 to a liquid culture medium for culture, and obtaining the liquid microbial inoculum after the culture is finished.
The invention also provides a solid microbial inoculum which comprises the Geobacillus lituunicus G6-1 and a carrier.
The invention also provides a preparation method of the solid microbial inoculum, which comprises the following steps: and (2) mixing the liquid microbial inoculum and a carrier according to the mass ratio of 10.
The invention also provides application of the Geobacillus lituunicus G6-1 in aerobic composting of biogas residues.
The invention also provides a biogas residue aerobic composting method, which comprises the following steps:
1) Mixing the biogas residues and auxiliary materials to obtain a mixed material;
2) Inoculating the liquid microbial inoculum or the solid microbial inoculum into the mixed material obtained in the step 1) for aerobic composting.
As mentioned above, the microbial strain for high-temperature aerobic composting of biogas residues and the application thereof have the following beneficial effects:
(1) The Geobacillus lituanicus G6-1 strain has the characteristics of high salt tolerance and high NaCl tolerance concentration, and is suitable for raw materials with high salt content in kitchen waste anaerobic fermentation biogas residues.
(2) The Geobacillus lituunicus G6-1 disclosed by the invention is high-temperature resistant, strong in aerobic fermentation self-heating capacity, capable of enabling the maximum temperature of materials to reach 75 ℃, capable of prolonging the high-temperature period, quickly decomposing organic matters, accelerating the biogas residue composting process, improving the material maturity and shortening the composting time.
In conclusion, the Geobacillus lituunicus G6-1 can be applied to high-temperature aerobic composting of biogas residues, and has the advantages of high treatment efficiency, good economic benefit, convenience in operation and no pollution.
Drawings
FIG. 1 is a phylogenetic tree map of strain G6-1;
FIG. 2 shows the temperature change of a compost body in the process of applying the strain G6-1 to biogas residue high-temperature aerobic composting;
FIG. 3 shows the change of C/N of the strain G6-1 in the high-temperature aerobic composting process of the biogas residues;
FIG. 4 shows the change of the Germination Index (GI) of the seeds of the strain G6-1 applied to the biogas residue high-temperature aerobic composting process.
Detailed Description
The invention provides a biogas residue high-temperature aerobic composting microbial strain which is identified as Geobacillus lituanicus, is preserved in China Center for Type Culture Collection (CCTCC), has the strain name of Geobacillus lituanicus G6-1, the preservation date of 2022.1.5 and the preservation number of CCTCC NO: m2022022027, the preservation address is Wuhan university, wuhan, china.
The Geobacillus lituunicus G6-1 contains a gene sequence shown as SEQ ID No. 1.
The Geobacillus lituanicus G6-1 is a rod-shaped gram-positive bacterium; after culturing on a CYS solid culture medium plate for 24 hours, colonies with the diameter of 4.4-5.8 mm can be formed, and the colony morphology is yellow brown round, convex, opaque and glossy.
CYS solid medium: 3g/L of peptone, 2g/L of yeast extract, 1g/L of soluble starch, 3g/L of NaCl and MgCl 2 0.125g/L、CaCl 2 0.5g/L、FeSO 4 ·7H 2 0.01g/L of O, trace element solution (Na) 2 MoO 4 12g/L、MnCl 2 5g/L、ZnSO 4 ·7H 2 O 0.6g/L、CuSO 4 ·5H 2 O 0.15g/L、CoCl 2 ·6H 2 O 8g/L、NiCl 2 ·6H 2 O0.2 g/L) 100 mu L/L and 17g/L agar, and the pH value is adjusted to 7-7.5.
The Geobacillus lituunicus G6-1 can resist high temperature. Specifically, the maximum growth temperature of Geobacillus lituanicus G6-1 of the Geobacillus lituranicus is 75 ℃. Above 75 ℃ the strain does not grow.
The Geobacillus lituunicus G6-1 salt-resistant Paenibacillus immediately. The maximum salt tolerance concentration of the Geobacillus lituranicus G6-1 of the Geobacillus lituunicus lituus is 80G/L sodium chloride. The highest salt tolerance concentration of the Geobacillus lituunicus G6-1 of Geobacillus lituunicus is 80G/L, wherein the sodium chloride refers to the following components:
1) Inoculating Geobacillus lituanicus G6-1 to a plate of a CYS solid culture medium containing 80G/L NaCl, and culturing for 24h at 50 ℃;
2) After 24 hours, the Geobacillus lituunicus G6-1 grows and breeds normally, namely the highest salt tolerance concentration of the Geobacillus lituunicus G6-1 is considered to be 80G/L of sodium chloride.
In one embodiment, geobacillus lituanicus G6-1 can be prepared into a liquid microbial agent or a solid microbial agent to be added into a mixed material consisting of biogas residues and auxiliary materials for aerobic composting.
The biogas residue refers to solid substances left after anaerobic fermentation of organic substances, and mainly comprises refractory cellulose and a large amount of anaerobic microorganisms.
When the Geobacillus lituranicus G6-1 is used for aerobic composting of biogas residues, the highest composting temperature is 75 ℃. When the Geobacillus lituunicus G6-1 is used for aerobic composting of the biogas residues, the relative high-temperature maintenance period is 5 days. The time taken to reach the maximum heap temperature was shortened by 3 days compared to the control.
The highest temperature of the pile body is the highest temperature which can be reached by the pile body when the aerobic composting is carried out on the biogas residues by using the Geobacillus lituunicus G6-1.
The high temperature maintenance period refers to the days for which the temperature of the stack reaches more than 55 ℃. The relative high temperature maintenance period refers to days in which the high temperature maintenance period of the experimental group is prolonged compared with that of the control group.
When the Geobacillus lituunicus G6-1 is used for aerobic composting of the biogas residues, the germination index of the seeds is 104 percent in 30 days.
When the Geobacillus lituunicus G6-1 strain of Geobacillus lituunicus is used for aerobic composting of biogas residues, the reduction rate of the relative carbon-nitrogen ratio (C/N) in 30 days is 23.49%.
The maximum heap body temperature, the relative high temperature maintenance period, the 30-day relative carbon-nitrogen ratio reduction rate and the 30-day seed germination index are obtained through the following steps:
1) Mixing biogas residues obtained by anaerobic fermentation of kitchen waste with rice straws according to the mass ratio of 10 to 3 to obtain a mixed material, and mixing a solid microbial inoculum of Geobacillus lituunicus G6-1 of Geobacillus immediately with the mixed material, wherein the mass of the solid microbial inoculum is 0.5 percent of the mass of the mixed material;
2) Fermenting in aerobic fermentation equipment, wherein the materials are stirred once a day in the fermentation process, and the ventilation rate is 20L/(kg.min);
3) During the fermentation process, the temperature of the center of the pile body is measured at the same time point every day, and the carbon-nitrogen ratio (C/N) and the Germination Index (GI) of the seeds are sampled and determined. The operation of the control group is completely the same except that the solid microbial inoculum is not added;
the carbon to nitrogen ratio is calculated by the following formula:
carbon to nitrogen ratio = TOC/TN
Wherein, the TOC of the material is measured by a TOC analyzer by a combustion method, and the TN is measured by a Kjeldahl method.
The carbon-nitrogen ratio decrease rate is calculated by the following formula:
carbon-nitrogen ratio decrease rate = (C/N) 0 -C/N i )/(C/N 0 )×100％
Wherein C/N 0 The initial carbon-nitrogen ratio, C/N, of the material compost i The carbon-nitrogen ratio of the materials to be composted on the ith day.
Relative carbon to nitrogen ratio reduction rate = carbon to nitrogen ratio reduction rate of experimental group on day i-carbon to nitrogen ratio reduction rate of control group on day i
The 30-day relative carbon-nitrogen ratio decrease rate = the carbon-nitrogen ratio decrease rate of the experimental group on day 30-the carbon-nitrogen ratio decrease rate of the control group on day 30.
The second aspect of the invention provides a liquid microbial agent, which comprises the Geobacillus lituus G6-1, wherein the concentration of the Geobacillus lituus G6-1 in the liquid microbial agent is at least 1 x 10 8 CFU/mL。
The third aspect of the invention provides a preparation method of the liquid microbial inoculum, which comprises the following steps: inoculating the pure strain of Geobacillus lituranicus G6-1 to a liquid culture medium for culture, and obtaining the liquid microbial inoculum after the culture is finished.
In one embodiment, the pure strain of Geobacillus lituunicus G6-1 is inoculated into a liquid culture medium for culture, then the culture solution is inoculated into another liquid culture medium for amplification culture, and the liquid microbial inoculum is obtained after multiple amplification culture.
In one embodiment, the temperature of the culture is 40 to 60 ℃. Specifically, the culture temperature is, for example, 45 to 55 ℃ and 47 to 53 ℃.
In one embodiment, the stirring speed for the culture is 150 to 200r/min.
In one embodiment, the pure strain of Geobacillus lituunicus G6-1 is inoculated into a liquid culture medium for culture for 13-18 h and then inoculated into another liquid culture medium for expansion culture.
In one embodiment, the liquid microbial inoculum of the strain can be obtained after 2-6 times of expansion culture.
In one embodiment, the liquid medium is a CYS liquid medium.
The CYS liquid culture medium comprises the following components: 3g/L of peptone, 2g/L of yeast extract, 1g/L of soluble starch, 3g/L of NaCl, and MgCl 2 0.125g/L、CaCl 2 0.5g/L、FeSO 4 ·7H 2 0.01g/L of O, trace element solution (Na) 2 MoO 4 12g/L、MnCl 2 5g/L、ZnSO 4 ·7H 2 O 0.6g/L、CuSO 4 ·5H 2 O 0.15g/L、CoCl 2 ·6H 2 O 8g/L、NiCl 2 ·6H 2 O0.2 g/L) 100 mu L/L, and the pH value is adjusted to 7-7.5.
The invention provides a solid microbial inoculum, which comprises Geobacillus lituunicus G6-1 and a carrier.
In one embodiment, the support is a solid support. The solid phase carrier is selected from one or more of soluble starch, cyclodextrin, wood dust, rice hulls and straws.
In one embodiment, the water content of the solid microbial inoculum is 10% or less.
The fifth aspect of the invention provides a preparation method of a solid microbial inoculum, which comprises the following steps: and (2) mixing the liquid microbial inoculum with a carrier according to the mass ratio of 10 to 1.
In one embodiment, the mass ratio of the liquid microbial inoculum to the carrier is 7.
In one embodiment, the mixture is dried until the water content of the mixture is 10% or less.
The sixth aspect of the invention provides application of Geobacillus lituranicus G6-1 in high-temperature aerobic composting of biogas residues.
The high-temperature aerobic compost is selected from any one of a closed type, a strip stack type and a groove type.
The seventh aspect of the invention provides a method for high-temperature aerobic composting of biogas residues, which comprises the following steps:
1) Mixing the biogas residues and auxiliary materials to obtain a mixed material;
2) Mixing the solid microbial inoculum or the liquid microbial inoculum with the mixed material obtained in the step 1) and then carrying out high-temperature aerobic composting.
In the step 1), the auxiliary materials are selected from one or more of straws, rice husks, wheat bran, sawdust, edible fungus residues, garden garbage and livestock and poultry manure.
The mass of the auxiliary material is 10-30% of the mass of the biogas residue.
In one embodiment, the initial moisture content of the mixed material is 55-65%; the initial pH is 7.0-7.5; the initial C/N is 25-30.
In one embodiment, the addition amount of the solid microbial inoculum or the liquid microbial inoculum is 0.1-10% of the mass of the mixed material.
In one embodiment, the aeration and stirring are performed during the high-temperature aerobic composting process. Preferably, the materials are periodically ventilated and stirred or turned over once in 1 to 3 days. The ventilation stirring can ensure that the temperature of the materials is uniform and the materials are carried out under aerobic conditions.
The fermentation time of the high-temperature aerobic compost can be specifically determined according to different materials, and the fermentation can be stopped after the germination index of the seeds reaches the standard. In one embodiment, the high temperature aerobic composting fermentation time is 10 days or more. The aerobic fermentation time is, for example, 10 to 15 days, 15 to 20 days, 20 to 25 days or longer.
The following embodiments of the present invention are provided by way of specific examples, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. 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.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not to be limited to the specific embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, the invention may be practiced using any method, device, and material that is similar or equivalent to the methods, devices, and materials described in examples herein, in addition to those described in prior art practice and the description herein.
Example 1: screening and functional verification of biogas residue high-temperature aerobic compost microbial strains
The screening and culture of the strain adopt CYS liquid culture medium and solid culture medium, and the components are as follows:
CYS liquid medium: 3g/L of peptone, 2g/L of yeast extract, 1g/L of soluble starch, 3g/L of NaCl and MgCl 2 0.125g/L、CaCl 2 0.5g/L、FeSO 4 ·7H 2 0.01g/L of O, trace element solution (Na) 2 MoO 4 12g/L、MnCl 2 5g/L、ZnSO 4 ·7H 2 O 0.6g/L、CuSO 4 ·5H 2 O 0.15g/L、CoCl 2 ·6H 2 O 8g/L、NiCl 2 ·6H 2 O0.2 g/L) 100 mu L/L, and the pH value is adjusted to 7-7.5.
CYS solid medium: 17g/L agar was added on the basis of CYS liquid medium.
The prepared culture medium is sterilized by high pressure steam at 121 deg.C for 20 min.
Separating and screening strains: the strains were isolated by dilution plating. The experimental sample is collected from a pile of high-temperature compost, 1g of the sample is weighed into a conical flask filled with 25mL of sterile water, and the conical flask is placed in a shaking table at 50 ℃ and 150r/min for 1h, so that the strains are uniformly dispersed in the liquid. Diluting the liquid to prepare a series of diluted liquids in sequence. Properly selecting 3-4 dilution concentrations, using a pipette to divide 100 mu L of the dilution to inoculate on a CYS solid culture medium, using a sterilized coating rod to uniformly coat the dilution, and placing the dilution in an incubator at 50 ℃ for culturing for 16-24 h. According to the colony morphology on the plate, colonies with different morphologic sizes are picked, and the colonies are subjected to streak purification and then are numbered and preserved. A total of 7 strains were obtained by separation and purification.
And (4) function verification: inoculating the obtained pure strain into a CYS liquid culture medium, and culturing for 16h at 50 ℃ under the condition of 180 r/min. The strain is inoculated into a mixture of 5kg of biogas residues and 1kg of wood chips according to the inoculation amount of 5% for aerobic composting function verification, and the result shows that the seed Germination Index (GI) is the highest after the biogas residues and the wood chips mixed material inoculated with the strain G6-1 are composted, and reaches 101% after 20 days.
Gram staining and microscopic observation of the strain G6-1 showed that it was a gram-positive bacterium, rod-like. After culturing on a CYS solid culture medium plate for 24 hours, colonies with the diameter of 4.4-5.8 mm can be formed, and the colony morphology is yellow brown round, convex, opaque and glossy.
Temperature tolerance determination of Strain G6-1: streaking the strain G6-1 on a CYS solid culture medium plate, respectively placing in incubators with the temperatures of 50, 55, 60, 65, 70, 75 and 80 ℃, recording the growth condition of the strain, and judging the highest tolerance temperature of the strain. As a result, it was found that the maximum tolerated temperature of the strain G6-1 was 75 ℃.
And (3) determining the salt tolerance of the strain G6-1: inoculating the strain G6-1 to CYS solid culture medium plates with different NaCl concentrations (20-120G/L) by a scribing method, carrying out aerobic culture in an incubator at 50 ℃ for 24h, and regularly observing the growth condition of the strain, wherein the result shows that the strain G6-1 grows well on the CYS solid culture medium containing 80G/L NaCl, the colony size is 3-4 mm, and the strain G6-1 does not grow on the CYS solid culture medium containing 90G/L NaCl, which indicates that the highest salt tolerance concentration of the strain G6-1 for good growth is 80G/L NaCl.
Example 2: identification of Strain G6-1
Genomic DNA of the strain G6-1 was extracted and used as a template to amplify the strain 16S rDNA using a pair of universal primers (27F, 1492R). The upstream primer is 27F (5-. The PCR reaction (20. Mu.L) was as follows: 0.5 μ L of template DNA, 10 μ L of PCR Taqmix, 0.6 μ L of each of the upstream and downstream primers, and ddH 2 O to the reaction system was 20. Mu.L. PCR procedure: pre-denaturation at 94 ℃ for 5min, denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s, extension at 72 ℃ for 1min for 30s, circulating for 30 times, extension at 72 ℃ for 10min, and storing at 4 ℃. Purification and sequencing of the PCR product was performed by Shanghai Jili Biotechnology Ltd. The 16S rDNA sequence obtained by sequencing was submitted at NCBI, and homology sequence alignment analysis was performed with GenBank by software, and MEGA11 software was used to construct phylogenetic trees of the strain (as shown in FIG. 1).
The 16S rDNA length of the strain G6-1 obtained by sequencing is 1403bp, and the nucleotide sequence is shown as SEQ ID NO. 1. The sequence has 99% similarity with Geobacillus lituunicius (NCBI serial number: NR 025657.1) in NCBI database by Nucleotide BLAST analysis. Simultaneously combining the morphological characteristics and growth characteristics of the bacterium: gram positive, rod-like; after the culture is carried out on a CYS solid culture medium plate for 24 hours, colonies with the diameter of 4.4-5.8 mm can be formed, and the colonies are in a yellow brown circular convex shape, are opaque and have luster. Finally, the screened G6-1 strain is determined to be Geobacillus lituus of Geobacillus lituus, and the strain is named as Geobacillus lituus G6-1.
Example 3: preparation of bacterial strain G6-1 microbial inoculum
Preparing a liquid microbial inoculum: inoculating G6-1 pure strain into 5mL CYS liquid culture medium, culturing for 16h in a constant temperature shaking table at 50 ℃ and 180r/min, then inoculating the pure strain into the same liquid culture medium by an inoculum size of 1% in volume ratio, and performing multistage amplification culture under the same condition to obtain the liquid microbial inoculum of the strain, wherein the concentration of Geobacillus lituranicus G6-1 is 1 x 10 8 CFU/mL。
Preparing a solid microbial inoculum: mixing a liquid microbial agent and carrier soluble starch according to a mass ratio of 5 to 1, and performing spray drying until the water content of the mixture is below 10% to obtain a solid microbial agent, wherein the concentration of Geobacillus lituus G6-1 is 1 × 10 8 CFU/mL。
Example 4: strain G6-1 strengthens high-temperature aerobic composting effect of biogas residues
10kg of biogas residues obtained by anaerobic fermentation of kitchen waste and 3kg of rice straws are placed in aerobic fermentation equipment with good heat preservation performance, 13kg of mixed materials are obtained after uniform mixing, 65G of solid microbial inoculum of Geobacillus lituanicus G6-1 of Geobacillus lituanicus is added according to the mass ratio of 0.5%, and the mixture is uniformly stirred. At this time, the initial C/N of the material was 26, the initial water content of the material was adjusted to 60%, and the initial pH was 7.0. Then starting aerobic fermentation of the materials, continuously ventilating in the process, and stirring the materials once a day. Meanwhile, a control group without added microbial inoculum is arranged, and aerobic composting is carried out by adopting the same materials and conditions. And measuring the temperature of the compost at the same time every day, sampling and measuring other physical and chemical indexes, and comparing the change conditions of the indexes in the composting process of the experimental group and the control group. The detection method of the germination index of the seeds in the physical and chemical indexes is carried out according to the regulation in organic fertilizer (NY 525-2021).
The stack temperature change is shown in figure 2. As can be seen from FIG. 2, the stack temperatures of both groups showed a tendency of rising first and then falling, and the temperature variation range was between 32 ℃ and 75 ℃. The initial temperature of the two groups of piles is about 32 ℃, after the experimental group is added with the Geobacillus lituuicus G6-1 solid microbial inoculum, the temperature of the piles is rapidly increased, the highest temperature is 75 ℃, and the time required for reaching the highest temperature is 6d; the maximum temperature of the control group was 62 ℃, the time required for reaching the maximum temperature was 9d, and the time required for reaching the maximum temperature of the experimental group was 3d shorter than that of the control group. In the composting process, the maintaining time of the experimental group in the high temperature period (not less than 55 ℃) is 8 days, the maintaining time of the control group in the high temperature period is 3 days, and the maintaining time of the experimental group in the high temperature period is prolonged by 5 days compared with that of the control group. The result shows that the inoculation of Geobacillus lituunicus G6-1 microbial inoculum of Geobacillus lituunicus can improve the highest temperature of a pile body in the aerobic composting process of biogas residues and prolong the duration of a high temperature period.
The C/N variation is shown in FIG. 3. The change trend of C/N in the two groups of stacks is continuously reduced, the C/N of a control group is reduced to 20.5 from 25.6, and the C/N of an experimental group added with Geobacillus lituranicus G6-1 microbial inoculum of Geobacillus lilacinus is reduced to 14.6 from 25.8. This is because the organic matter in the compost is continuously decomposed and utilized by microorganisms as the composting proceeds. Wherein, the C/N reduction rate in the experimental group added with the microbial inoculum is 23.49 percent higher than that in the control group, which shows that the microbial metabolism rate in the pile body of the experimental group is faster.
GI variation is shown in fig. 4. The germination indexes of the seeds of the two groups of piles are increased continuously, the initial GI is about 20%, and the germination indexes of the seeds of the control group are increased to 42% on the 30 th day of composting; the GI of the experimental group added with the microbial inoculum reaches 72 percent at 18 days of composting, meets the index of germination index in organic fertilizer (NY 525-2021), and rises to 104 percent at 30 days to achieve complete decomposition. Therefore, the Geobacillus lituanicus G6-1 has the capability of fast decomposing, and can obviously promote the biogas residue compost fermentation.
After composting is finished, physicochemical indexes such as organic matters and total nutrients of the organic fertilizer products obtained by the two groups of experiments are measured, and the results show that: adding ofThe organic matter (calculated on a drying basis) of the experimental group organic fertilizer product of the microbial inoculum is 78 percent, and the total nutrient is (N + P) 2 O 5 +K 2 O) (on a dried basis) 8.7%; the organic matter (calculated on a drying basis) of the organic fertilizer product of the control group without adding the microbial inoculum is 65 percent, and the total nutrient is (N + P) 2 O 5 +K 2 O) (on a dried basis) was 7.3%. The results show that the organic matter and total nutrient content of the organic fertilizer products of the experimental group are higher than those of the control group, and the product quality is improved.
In conclusion, the Geobacillus lituus G6-1 microbial inoculum provided by the invention is added into aerobic biogas residue compost, so that the degradation of organic matters in the compost can be effectively promoted, the composting process is accelerated, and the maturity of the compost and the quality of organic fertilizers are improved.
The above examples are intended to illustrate the disclosed embodiments of the invention and are not to be construed as limiting the invention. In addition, various modifications of the methods set forth herein, as well as variations of the methods of the invention, will be apparent to those skilled in the art without departing from the scope and spirit of the invention. While the invention has been specifically described in connection with various specific preferred embodiments thereof, it should be understood that the invention is not limited to those specific embodiments. Indeed, various modifications of the above-described embodiments which are obvious to those skilled in the art to which the invention pertains are intended to be covered by the scope of the present invention.
Sequence listing
<110> Shanghai higher research institute of Chinese academy of sciences
<120> microbial strain for high-temperature aerobic composting of biogas residues and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1403
<212> DNA
<213> Bacillus Liotaoticus (Geobacillus lituranicus G6-1)
<400> 1
tgagggttac ctcaccgact tcgggtgttg caagctctcg tggtgtgacg ggcggtgtgt 60
acaaggcccg ggaacgtatt caccgcggca tgctgatccg cgattactag cgattccggc 120
ttcatgcagg cgagttgcag cctgcaatcc gaactgagag cggctttttg ggattcgctc 180
cccctcgcgg gttcgcagcc ctttgtaccg cccattgtag cacgtgtgta gcccaggtca 240
taaggggcat gatgatttga cgtcatcccc accttcctcc gacttgtcgc cggcagtccc 300
tctagagtgc ccaccttcgt gctggcaact agaggcgagg gttgcgctcg ttgcgggact 360
taacccaaca tctcacgaca cgagctgacg acaaccatgc accacctgtc accctgtccc 420
cccgaagggg gaacgcccaa tctcttgggt tgtcagggga tgtcaagacc tggtaaggtt 480
cttcgcgttg cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 540
ctttgagttt cagccttgcg gccgtactcc ccaggcggag tgcttatcgc gttagctgca 600
gcactaaagg gtgtgacccc tctaacactt agcactcatc gtttacggcg tggactacca 660
gggtatctaa tcctgtttgc tccccacgct ttcgcgcctc agcgtcagtt gcaggccaga 720
gagccgcctt cgccactggt gttcctccac atctctacgc atttcaccgc tacacgtgga 780
attccgctct cctctcctgc actcaagtcc cccagtttcc aatgaccctc cacggttgag 840
ccgtgggctt tcacatcaga cttaaggaac cgcctgcgcg cgctttacgc ccaataattc 900
cggacaacgc tcgcccccta cgtattaccg cggctgctgg cacgtagtta gccggggctt 960
cctcgtgagg taccgtcacc gcgccgccct cttcgaacgg cgctccttcg tccctcacaa 1020
cagagcttta cgacccgaag gccttcttcg ctcacgcggc gtcgctccgt caggctttcg 1080
cccattgcgg aagattccct actgctgcct cccgtaggag tctgggccgt gtctcagtcc 1140
cagtgtggcc ggtcaccctc tcaggccggc tacgcatcgt cgccttggtg agccgttacc 1200
tcaccaacta gctaatgcgc cgcgggccca tccgcaagtg acagccaaag gccgcctttc 1260
aaccaaagac catgcggtct tcggtgttat ccggtattag ctccggtttc ccggagttat 1320
cccggtcttg cgggcaggtt gcccacgtgt tactcacccg tccgccgctg accaaatcag 1380
agcaagctcc gatttggtcc gct 1403
Claims (8)
1. A Paeonia lactiflora (Bacillus licheniformis) (B.sp.) (C.sp.)Geobacillus lituanicus) G6-1 with the preservation number of CCTCC NO: M2022022027.
2. A liquid microbial agent comprising the Bacillus terroris (Geobacillus sp.) (according to claim 1)Geobacillus lituanicus) G6-1, bacillus terradoides (A) in the liquid microbial inoculumGeobacillus lituanicus) The concentration of G6-1 is at least 1X 10 8 CFU/mL。
3. The method for preparing a liquid microbial inoculum according to claim 2, comprising the steps of: bacillus Litahuanianus as claimed in claim 1 (A)Geobacillus lituanicus) Inoculating the pure strain of G6-1 into a liquid culture medium for culture, and obtaining the liquid microbial inoculum after the culture is finished.
4. A solid microbial agent comprising the Bacillus terroris (Geobacillus sp.) (claim 1)Geobacillus lituanicus) G6-1 and a carrier.
5. The method for preparing a solid microbial inoculum according to claim 4, which comprises the following steps: mixing the liquid microbial inoculum according to claim 2 with a carrier according to a mass ratio of 10 to 1.
6. The Bacillus taedania sp (A. Terrestris) (B. Terrestris) as claimed in claim 1Geobacillus lituanicus) G6-1 is applied to the aerobic composting of the biogas residues.
7. A biogas residue aerobic composting method is characterized by comprising the following steps:
1) Mixing the biogas residues and auxiliary materials to obtain a mixed material;
2) Inoculating the liquid microbial inoculum according to claim 2 or the solid microbial inoculum according to claim 4 into the mixed material obtained in the step 1) for aerobic composting.
8. The method of claim 7, further comprising one or more of the following conditions:
A. the auxiliary materials are selected from one or more of straws, rice hulls, wheat bran, sawdust, edible fungus residues, garden garbage or livestock and poultry manure;
B. the mass of the auxiliary material is 10 to 30 percent of the mass of the biogas residue;
C. the addition amount of the microbial inoculum is 0.1 to 10 percent of the mass of the mixed material;
D. the initial water content of the mixed material is 55 to 65 percent;
E. the initial pH of the mixed material is 7.0 to 7.5;
F. the initial C/N of the mixed material is 25 to 30.
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| CN109022327A (en) * | 2018-09-05 | 2018-12-18 | 福建省致青生态环保有限公司 | A kind of preparation method of microorganism mix bacterium agent and the application in During High-Temperature Composting |
| CN112725214A (en) * | 2020-10-23 | 2021-04-30 | 江西农业大学 | Biogas residue aerobic compost composite microbial agent and application thereof |
| CN112760274A (en) * | 2021-04-07 | 2021-05-07 | 中国科学院上海高等研究院 | Organic solid waste high-temperature aerobic composting strain and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109022327A (en) * | 2018-09-05 | 2018-12-18 | 福建省致青生态环保有限公司 | A kind of preparation method of microorganism mix bacterium agent and the application in During High-Temperature Composting |
| CN112725214A (en) * | 2020-10-23 | 2021-04-30 | 江西农业大学 | Biogas residue aerobic compost composite microbial agent and application thereof |
| CN112760274A (en) * | 2021-04-07 | 2021-05-07 | 中国科学院上海高等研究院 | Organic solid waste high-temperature aerobic composting strain and application thereof |
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