CN105567609B - One plant of high temperature resistant garden waste decomposer ST2 and its application - Google Patents

One plant of high temperature resistant garden waste decomposer ST2 and its application Download PDF

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CN105567609B
CN105567609B CN201610119770.7A CN201610119770A CN105567609B CN 105567609 B CN105567609 B CN 105567609B CN 201610119770 A CN201610119770 A CN 201610119770A CN 105567609 B CN105567609 B CN 105567609B
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temperature
composting
bacterium solution
adsorbent
garden waste
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CN105567609A (en
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彭霞薇
连鹏
周金星
郑景明
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Beijing Forestry University
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/01Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465Streptomyces
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • C12N1/205Bacterial isolates
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00Other organic fertilisers
    • C05F11/08Organic fertilisers containing added bacterial cultures, mycelia or the like
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F3/00Fertilisers from human or animal excrements, e.g. manure
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20Bacteria; Culture media therefor
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00Technologies for solid waste management
    • Y02W30/40Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Abstract

The invention discloses one plant of thermophilic streptomyces diastaticus of high-temperature fibre element degradation bacteria (Streptomyces thermodiastaticus), deposit number is CGMCC No.12134.Bacterium of the present invention a large amount of producing enzymes, cellulase activity height can have high temperature resistant, efficient degradation cellulosic nature in 40-70 DEG C of high temperature, can be applied in During High-Temperature Composting system as microbial bacterial agent, be suitable for garden waste compost.

Description

One plant of high temperature resistant garden waste decomposer ST2 and its application
Technical field
The invention belongs to field of environmental biotechnology, and in particular to the plant height effect filtered out from garden waste compost The thermoduric bacteria of degraded cellulose and its application in garden waste During High-Temperature Composting.
Background technique
Garden waste refers to that ornamental plant withers and falls naturally or manually trim generated deadwood, fallen leaves, grass cuttings, residual flower, tree Wood and shrub beta pruning and other plant residues etc., main component are cellulose and hemicellulose difficult to degrade.In recent years, China garden The speed increase of the annual 8%-10% of woods waste brings large drag forces to City Green process, and China handles gardens at present The mode of waste predominantly burn and fill up, both processing modes not only waste renewable resource also create it is serious Environmental pollution.Therefore, how garden waste is rationally effectively treated, making its resource utilization is the heat of everybody current common concern One of point problem.
Organic fertilizer and seedling medium etc. is made by garden waste is decomposed using composting technology, is its resource utilization One of effective outlet, however lead to heap fertilizer efficiency containing substances difficult to degrade such as a large amount of lignin, celluloses in garden waste Rate is low, the period is long, greatly limits the recycle value of garden waste.Therefore it needs that specific micro- life is added into compost Object, to accelerate garden waste digest process.Cellulose-degrading bacteria is that one kind can generate extracellular cellulase, and cellulose is big Molecule is hydrolyzed into the microorganism of glucose, can be with the degradation speed of accelerating fibers cellulosic material.In nature, the micro- of cellulose is generated There are many biological species, and study and apply at present more is fungi, such as trichoderma, Penicillium, aspergillus, rhizopus.Bacterium Research and application with actinomyces is less.Cellulose degradation occurs mainly in the megathermal period in composting process, and fungi mainly exists Under medium temperature condition, enzyme activity is maximum, and with the rising of composting process temperature, the enzymatic activity of fungi number of viable and its generation drops significantly Low, which limits the utilizations in compost of cellulase-producing mould.The screening of high-temperature fibre element degradation bacteria and application are The effective measures to solve the above problems.
Many cellulose-degrading bacterias both for agricultural wastes such as corn stover, straw, wheat straws, it is few specifically for The screening of the degradation bacteria of garden waste cellulosic material and research on utilization.Chinese patent " a plant height temperature cellulose-degrading bacteria and It is applied " it (number of patent application: 201410018582.6) discloses one plant and is separated from garden waste megathermal period compost sample High temperature fiber element degradation bacteria ground bacillus, cellulase activity be 7.8 U/ml, the bacterium be bacterium.About discarded from gardens The report that high-temperature fibre element degradation actinomyces are separated in object compost is less.
Summary of the invention
The technical problems to be solved by the present invention are: providing one plant of thermophilic streptomyces diastaticus of high temperature resistant, which can be in 40- A large amount of producing enzymes in 70 DEG C of high temperature, cellulase activity is high, has high temperature resistant, efficient degradation cellulosic nature, can be as micro- Bacteria agent is applied in During High-Temperature Composting system, is suitable for garden waste compost.
Present invention provide the technical scheme that one plant of thermophilic streptomyces diastaticus of high-temperature fibre element degradation bacteria (Streptomyces thermodiastaticus) ST2, deposit number is CGMCC No.12134, is preserved in China Microbiological Culture presevation administration committee common micro-organisms center.
Thermophilic streptomyces diastaticus of the present invention (Streptomyces thermodiastaticus) ST2 is in 2016 Years 18 days 2 months for the preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC institute (preservation address is: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode: 100101), deposit number is CGMCC No.12134, through detecting Survival.
The thermophilic streptomyces diastaticus of the present invention (Streptomyces thermodiastaticus) ST2, it is from Bei Jingchang The strain separated in the sample of flat area apple orchard garden waste compost high temperature mid-term acquisition, being numbered is bacterial strain ST2, should Bacterium can generate cellulase and degraded cellulose under the conditions of 40-70 DEG C.Bacterial strain on Gao Shi I culture medium when growing, gas Raw mycelia is light grey, mycelia multiple-limb, and substrate mycelium lark does not generate soluble pigment, and aerial hyphae branch is raw in gas There is fibrillae of spores on mycelia.Under microscope, thallus is in mycelioid, and Gram's staining is positive, and spore oval, surface is smooth, The slightly curved song of fibrillae of spores, in the shape of a spiral.It is sent out in conjunction with bacterium colony morphological features with the system based on bacterial 16 S rDNA gene order Educate analysis, be accredited as thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus)。
The method of bacterial strain of the present invention production cellulase, by the bacterium be inoculated in fermentation medium (medium component: CMC-Na 0.5%, peptone 1%, wheat bran 3%, NaCl 2%, KH2PO40.1%g, MgSO4•7H2O 0.02%, (NH4)2SO4 0.3%, pH 7.0-7.6) in, turn to cultivate 3-4d in shaking table in temperature 50 C, rpm150, centrifuging and taking supernatant, measures cellulose Enzymatic activity.
It is the During High-Temperature Composting decomposing agent of main composting material that the present invention also provides a kind of suitable for garden waste, this is decomposed Agent is obtained in a manner of liquid fermentation, and bacterial concentration is 5 × 109CFU/mL, wheat bran and maize flour 2:1 mixing are adsorbed as bacterium solution Agent is mixed with adsorbent 1:1 by bacterium solution, is fabricated to solid-state microbial inoculum.
Meanwhile the present invention also provides the thermophilic streptomyces diastaticus ST2 bacterial strains using garden waste as main material Compost maturity in application add animal wastes and water with garden waste for main composting material, make mixed material carbon nitrogen Than for 25-40:1, moisture content 50-60%, solid-state decomposing agent is inoculated into compost material in 5% ratio of weight of material, carry out high Warm compost.
The invention has the following advantages:
Thermophilic streptomyces diastaticus of the present invention (Streptomyces thermodiastaticus) ST2 is from garden The strain of degraded cellulose is filtered out in woods castoff compost, and more cellulose can be generated in 40-70 DEG C of temperature range Enzyme, enzyme activity are up to 63.55 U/mL, have high temperature resistant, efficient degradation cellulosic nature.Opposite fungi enzyme activity under mesophilic condition Maximum, and the problem that bacterium producing enzyme vigor is lower, thermophilic streptomyces diastaticus had not only adapted to hot environment, but also can generate higher Cellulase has good degradation capability to cellulosic material, so thermophilic streptomyces diastaticus is suitable for composting process Complex environment.
Solid-state decomposing agent is made in the high-temperature fibre element degradation bacteria that the present invention obtains to be added to based on garden waste Want in the compost of material, compared with the control not being inoculated with, can be improved compost enter the megathermal period time, extend the megathermal period continue Time megathermal period temperature reduces composting C/N ratio, to accelerate compost maturity process.It can be answered as microbial bacterial agent For being suitable for garden waste compost in During High-Temperature Composting system.
The present invention be directed to the decomposing agents of the strain of garden waste composting material screening and preparation, can be discarded for gardens The resource utilization of object provides a reasonable, effective approach, realizes the purpose for reducing environmental pollution, resource circulation utilization.
Detailed description of the invention
Fig. 1 is thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus) isolate and purify picture
The thermophilic streptomyces diastaticus of Fig. 2 present invention (Streptomyces thermodiastaticus) 16S rDNA Gene order.
Fig. 3 by thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus) and close bacterial strain The gene order of 16SrDNA carries out the phylogenetic tree picture constructed when homology Blast is compared.
Heap temperature changes in Fig. 4 composting process.
Carbon-nitrogen ratio (C/N ratio) changes in Fig. 5 composting process.
Specific embodiment
Come that the present invention is furture elucidated below by the detailed description of specific embodiment, but is not to limit of the invention System, only illustrates.
The screening of 1 high-temperature fibre element degradation bacteria strains of embodiment
From Changping County, Beijing area apple orchard garden waste compost high temperature initial stage, high temperature mid-term, high temperature post material, Beijing Sample is acquired in Yanqing Plain afforestation afforestation waste deposit;Above-mentioned fresh sample 10g is weighed to be put in equipped with 10 Grain bead simultaneously fills in the conical flasks of 90 ml sterile waters, is placed in the shaking table of 30 DEG C of 150 rpm and shakes 30 min, makes Sample sufficiently scatters, and stands enrichment culture for 24 hours at 50 DEG C.1 ml supernatant is drawn with aseptic straw to be added to containing 9ml In the test tube of sterile water, this is 10-1Sample diluting liquid, then from 10-11ml is taken to be added in 9ml sterile water in sample, this is It is 10-2Sample diluting liquid, and so on, obtain 10-3、10-4、10-5、10-6Then sample diluting liquid draws 100 with pipettor The 10 of μ l-3、10-4、10-5、10-6 Sample diluting liquid is in (culture medium composition are as follows: K on Cellulose and congo red differential medium2HPO4 0.5g, microcrystalline cellulose 1.88g, MgSO4 0.25g, gelatin 2.0g, Congo red 0.5g, agar 16g, distilled water 1000ml, pH 7.0), dilution is spread evenly across entire plate with spreader, is placed in 50 DEG C of incubators and cultivates 3 days. The bacterium colony for having obvious transparent circle on the Congo red plate of cellulose is selected, is numbered, then is crossed to isolate and purify repeatedly and be obtained Pure strain is obtained, the strain after separation is connected on inclined-plane, 4 DEG C of preservations carry out subsequent experimentals.
By the pure bacterial strain for being preserved in 4 DEG C be transferred to carboxymethyl cellulose culture medium (medium component: CMC-Na 15.0g, NH4NO31.0g, yeast extract 1.0g, MgSO4•7H2O 0.5g, KH2PO41.0g, distilled water 1000ml, agar 16g, pH 7.0) on plate, then activation culture at 50 DEG C is provoked the single colonie on plate and is transferred on the Congo red plate of cellulose, is placed in It being cultivated in 50 DEG C of incubators, colony diameter d and transparent loop diameter D is measured after 72h, calculates its ratio H, i.e. H=D/d, H value is bigger, It is stronger to be worth the larger ability for indicating the bacterial strain decomposition of cellulose.According to formation transparent circle on the Congo red identification culture medium of cellulose Size primarily determines its cellulase-producing activity.
By aforesaid operations, more plants of cellulose-degrading bacterias are obtained, wherein in Changping County, Beijing area apple orchard garden waste The strain separated in the sample of compost high temperature mid-term acquisition, is named as ST1, and the micro- life of China was deposited on 2 15th, 2016 Object culture presevation administration committee common micro-organisms center is referred to as CGMCC (unit address: the Chaoyang District, Beijing City North Star The institute 3 of West Road 1, Institute of Microorganism, Academia Sinica, postcode: 100101), and deposit number CGMCC No.12134.Optical microscope and Gram's staining identification are carried out to ST2 bacterial strain, are somebody's turn to do when being grown on Gao Shi I culture medium, Aerial hyphae is light gray, and mycelia multiple-limb, substrate mycelium lark does not generate soluble pigment, aerial hyphae branch, in gas There is fibrillae of spores on raw mycelia.Under microscope, thallus is in mycelioid, and Gram's staining is positive, spore oval, surface light Sliding, the slightly curved song of fibrillae of spores is shown in Fig. 1 in the shape of a spiral.
2 ST2 bacterial strain molecular biology identification of embodiment
Molecular Identification is carried out to the thermophilic streptomyces diastaticus that screening obtains, follow the steps below: picking screens bacterium The single colonie of strain is inoculated in liquid Gao Shi I culture medium, 30 DEG C, 120r/min shaking table shaken cultivation, is taken out in 2d Culture solution, 5000r/min centrifugation 1min take supernatant, and according to bacterial genomes DNA extraction kit, (Tiangeng biochemical technology has Limit company provides), extract bacterium colony DNA;Universal primer 27F and 1492R carries out PCR amplification to the DNA of bacteria of extraction; 27F sequence is 5 '-AGA GTT TGA TCC TGG CTC AG-3 ';1492R sequence is 5 '-AAG GAG GTG ATC CAG CCG CA-3′;PCR product is subjected to sequence, sequencing result BLAST in NCBI database carries out sequence point Analysis, and carry out tetraploid rice.
16S rDNA gene order (the referring to Fig. 2) length of Potsdam bacillus brevis is 1426bp, by gene sequence Column are submitted on Genbank, carry out tetraploid rice, are then used 6.0 Software on Drawing phylogenetic tree of MEGA, are seen Fig. 3, So that it is determined that the kind of bacterial strain.The result shows that the sequence and thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus) 16S rDNA gene order similarity be up to 99%, in combination with colony morphology characteristic, life Reason biochemical character, thallus microscopic features determine ST1 bacterial strain be thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus)。
The thermophilic streptomyces diastaticus growth measurement of embodiment 3
By thermophilic streptomyces diastaticus (Streptomyces thermodiastaticus) ST2 be inoculated into CMC liquid training Support (CMC-Na 15.0g, NH in base4NO31.0g, yeast extract 1.0g, MgSO40.5g, KH2PO41.0g, distilled water 1000mL), culture solution being taken every 2h, connection is continuous to be sampled to 48h, the OD600 value in each period is measured, using incubation time as abscissa, The OD600 value of each sample point is ordinate, draws the growth curve of the bacterium, the i.e. growth measurement of the bacterium.It can be with from measurement result Find out 0-8h be period of delay, 9 ~ 16h be logarithmic growth phase, 16 ~ be for 24 hours stationary phase, > 26h be decline phase.The bacterium of logarithmic phase Strain growth it is rapid, energetic therefore later enzymatic production experiment in, 12 hours fermentation liquids of Ying Xuanyong be seed liquor into Row inoculation.
The thermophilic streptomyces diastaticus cellulase-producing vitality test of embodiment 4
By the thermophilic streptomyces diastaticus ST2 of logarithmic phase by 1% inoculum concentration be inoculated into fermentation medium (medium component: CMC-Na 0.5%, peptone 1%, wheat bran 3%, NaCl 2%, KH2PO40.1%, MgSO4•7H2O 0.02%, (NH4)2SO4 0.3%), pH 7.0-7.6) in, turn to cultivate 3-4d in shaking table in temperature 50 C, rpm150, culture 8000r/min is centrifuged 5min, taking supernatant is crude enzyme liquid, measures cellulase activity with DNS method.Taking 1 mL supernatant, (blank is with 1 in test tube The replacement of ml distilled water), it is placed in 50 DEG C of water-baths, preheats 2min, the substrate that 4 ml have been preheated at 50 DEG C is then added Solution takes out after clock reaction 5min, and 4mL DNS developing solution is added, is placed in boiling water bath after shaking up and heats 5 min, taken It is immediately placed in cooling in cold bath after out, is settled to 20ml with distilled water, measures absorbance after mixing at 540 nm wavelength. Convert the producing enzyme vigor of the bacterial strain after reference standard curve.Enzyme activity unit is according to international unit stipulative definition, i.e., in 1 mL In system, enzyme amount needed for catalyzing cellulose hydrolysis generates 1 μm of ol glucose in 1 min is an enzyme activity unit (U/mL).The cellulase activity of ST2 bacterial strain is 63.55 U/mL after measured.
The preparation of 5 solid-state decomposing agent of embodiment
(1) bacterial strain activates: take 4 DEG C of preservation inclined plane inoculatings of microorganism of the present invention to high family name I solid plate culture medium, 12h is cultivated in 50 DEG C of permanent case realizes bacterial strain activation.
(2) prepared by seed liquor: strain plate 1 in step (1) through slant activation is seeded to the sterile Gao Shi of 1 L I In fluid nutrient medium, seed liquor is obtained after 12 h are cultivated under the conditions of 50 DEG C of 150rpm shaking tables.
(3) preparation of fermentation seed liquid: above-mentioned seed liquor is by 6-10%(v/v) inoculum concentration be seeded to sterilized hair In fermentation tank, expansion fermented and cultured is carried out.Under conditions of temperature 50 C, 120 rpm of frequency of oscillation, it must be sent out after cultivating 48h Ferment seed liquor.
(4) preparation of solid-state decomposing agent: being used as bacterium solution adsorbent after wheat bran is mixed with maize flour according to 2:1 mass ratio, With bacterium solution obtained in step (3), is mixed by bacterium solution with adsorbent volume mass ratio 1:1, be fabricated to solid-state decomposing agent, bank up 1 Zhou Hou can be used for During High-Temperature Composting.
The compost effect test of 6 decomposing agent of embodiment
With garden wastes for main composting material, animal wastes and water are added, mixed material carbon-nitrogen ratio 25-40 is made: 1, moisture content 50-60%, solid-state decomposing agent are inoculated into compost material in 5% ratio of weight of material, During High-Temperature Composting are carried out, with not Add the material of decomposing agent for control, when heap temperature rises to 50 DEG C, start turning, the megathermal period, every turning in 2 days was primary, drop The warm phase, turning was primary weekly, not in turning after temperature drops to 40 DEG C.In composting process, pass through the daily temperature of measurement heap body Degree variation, composting material carbon nitrogen (C/N) investigate influence of the addition decomposing agent to garden-waste compost decomposition progress than variation.
Heap temperature variation is as shown in Figure 4 in composting process.As shown in Figure 4, the compost treatment of decomposing agent is inoculated in compost 2d temperature rises to 50 DEG C or more, and 50 DEG C or more of megathermal period continues 25d, and temperature is begun to decline later.And it does not connect The compost treatment temperature of kind just rises to 50 DEG C or more in compost 3d, and it is high to postpone 50 DEG C of 1d arrival or more than the processing of inoculation Wen Qi, and 50 DEG C or more duration megathermal period are 20d, fewer than the processing of inoculation 5d, is inoculated with the heap body megathermal period of processing Maximum temperature also above processing is not inoculated with, illustrate to accelerate compost after being inoculated with decomposing agent and enter time megathermal period, Yi Jigao Warm duration phase.C/N variation is as shown in Figure 5 in composting process.As shown in Figure 5, with the progress of compost, be inoculated with decomposing agent and The lasting processing decline degree for reducing, and being inoculated with of the processing C/N ratio not being inoculated with, which is higher than, is not inoculated with processing, thus illustrates, adds Decomposing agent made from this bacterium can accelerate composting process, improve the decomposed effect of garden waste compost.

Claims (5)

1. one plant of thermophilic streptomyces diastaticus of high-temperature fibre element degradation bacteria (Streptomyces thermodiastaticus), Deposit number is CGMCC No.12134.
2. it is a kind of suitable for garden waste be main composting material During High-Temperature Composting decomposing agent, it is characterised in that: the decomposing agent It is obtained in a manner of liquid fermentation, raw material is bacterium solution and adsorbent, and bacterium solution mixes in proportion with adsorbent, is fabricated to solid-state bacterium Agent, wherein bacterium solution be thermophilic streptomyces diastaticus described in claim 1 (Streptomyces thermodiastaticus) Bacterium solution, adsorbent are wheat bran and maize flour.
3. During High-Temperature Composting decomposing agent as claimed in claim 2, it is characterised in that: the bacterial concentration is 5 × 109CFU/mL, Wheat bran and maize flour 2:1 mixing are used as bacterium solution adsorbent, mix by bacterium solution with adsorbent 1:1, are fabricated to solid-state microbial inoculum.
4. application of the During High-Temperature Composting decomposing agent in compost maturity as described in Claims 2 or 3.
5. application as claimed in claim 4, it is characterised in that: with garden waste for main composting material, add animal excreta Just and water, make mixed material carbon-nitrogen ratio 25-40:1, moisture content 50-60%, solid-state decomposing agent presses the 2.5%-5% ratio of weight of material Example is inoculated into compost material, carries out During High-Temperature Composting.
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CN106350075A (en) * 2016-08-28 2017-01-25 何晓东 Soil improvement agent with sterilizing effect and preparation method of soil improvement agent
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CN114507090A (en) * 2021-12-28 2022-05-17 镇江贝思特有机活性肥料有限公司 Tobacco stem waste fermentation treatment device and treatment method

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