CN100465264C - Composite microorganism bacteria agent for compost fermentation and its producing method and use - Google Patents
Composite microorganism bacteria agent for compost fermentation and its producing method and use Download PDFInfo
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- CN100465264C CN100465264C CNB2006100785888A CN200610078588A CN100465264C CN 100465264 C CN100465264 C CN 100465264C CN B2006100785888 A CNB2006100785888 A CN B2006100785888A CN 200610078588 A CN200610078588 A CN 200610078588A CN 100465264 C CN100465264 C CN 100465264C
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- 238000000855 fermentation Methods 0.000 title claims abstract description 46
- 230000004151 fermentation Effects 0.000 title claims abstract description 46
- 241000894006 Bacteria Species 0.000 title claims abstract description 12
- 244000005700 microbiome Species 0.000 title claims description 34
- 239000002361 compost Substances 0.000 title claims description 17
- 238000000034 method Methods 0.000 title claims description 17
- 239000002131 composite material Substances 0.000 title claims 3
- 239000003795 chemical substances by application Substances 0.000 title abstract 5
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 45
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 45
- 241000235526 Mucor racemosus Species 0.000 claims abstract description 45
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 45
- 230000001580 bacterial effect Effects 0.000 claims abstract description 13
- 238000004519 manufacturing process Methods 0.000 claims abstract description 13
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 3
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 3
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 3
- 241000186660 Lactobacillus Species 0.000 claims description 42
- 229940039696 lactobacillus Drugs 0.000 claims description 42
- 239000000417 fungicide Substances 0.000 claims description 28
- 239000002068 microbial inoculum Substances 0.000 claims description 28
- 238000002156 mixing Methods 0.000 claims description 28
- 239000007788 liquid Substances 0.000 claims description 25
- 230000000855 fungicidal effect Effects 0.000 claims description 21
- 235000015097 nutrients Nutrition 0.000 claims description 21
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 18
- 229910052760 oxygen Inorganic materials 0.000 claims description 18
- 239000001301 oxygen Substances 0.000 claims description 18
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 17
- 229910052799 carbon Inorganic materials 0.000 claims description 17
- 230000008569 process Effects 0.000 claims description 15
- 238000003756 stirring Methods 0.000 claims description 13
- 239000002054 inoculum Substances 0.000 claims description 12
- 230000003068 static effect Effects 0.000 claims description 12
- 235000012204 lemonade/lime carbonate Nutrition 0.000 claims description 10
- 239000001888 Peptone Substances 0.000 claims description 9
- 108010080698 Peptones Proteins 0.000 claims description 9
- 235000013379 molasses Nutrition 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 238000012366 Fed-batch cultivation Methods 0.000 claims description 7
- -1 iginal Species 0.000 claims description 7
- 230000033228 biological regulation Effects 0.000 claims description 6
- 239000006052 feed supplement Substances 0.000 claims description 6
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims description 6
- 239000011368 organic material Substances 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 235000011187 glycerol Nutrition 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 abstract description 5
- 230000002906 microbiologic effect Effects 0.000 abstract 2
- 239000002028 Biomass Substances 0.000 abstract 1
- 238000012271 agricultural production Methods 0.000 abstract 1
- 238000005457 optimization Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 32
- 239000000463 material Substances 0.000 description 23
- 239000010902 straw Substances 0.000 description 11
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- 210000003608 fece Anatomy 0.000 description 8
- 230000001186 cumulative effect Effects 0.000 description 6
- 239000002994 raw material Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 235000015099 wheat brans Nutrition 0.000 description 6
- 241000209140 Triticum Species 0.000 description 5
- 235000021307 Triticum Nutrition 0.000 description 5
- 240000008042 Zea mays Species 0.000 description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000000354 decomposition reaction Methods 0.000 description 5
- 241000287828 Gallus gallus Species 0.000 description 4
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 4
- 239000010871 livestock manure Substances 0.000 description 4
- 235000009973 maize Nutrition 0.000 description 4
- 239000007858 starting material Substances 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
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- 238000012258 culturing Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000003895 organic fertilizer Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
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- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
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- 229910052757 nitrogen Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-BJUDXGSMSA-N oxygen-15 atom Chemical compound [15O] QVGXLLKOCUKJST-BJUDXGSMSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229940081969 saccharomyces cerevisiae Drugs 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 230000031068 symbiosis, encompassing mutualism through parasitism Effects 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention involves a compound microbiological bacterial agent and its production method and application. The bacteria agent contains Saccharomyces cerevisiae, Lactobacillus plantarum, Mucor racemosus, Aspergillus oryzae, through slant culture, first class seed, second class seed and combining fermentation to carry out the compound bacteria biomass and optimization of different strains matching, active viable count of compound bacteria agent is over 3*109cfu/mL. The compound microbiological bacterial agent can promote the digest rate of the organics efficiently, can be used in the production of bio-organics, and reduces agricultural production cost.
Description
Technical field
The present invention relates to a kind of complex micro organism fungicide, particularly a kind of complex micro organism fungicide that is used for organic material composting and produces biological organic fertilizer.The production method that the invention still further relates to described complex micro organism fungicide with and purposes in the organic materials compost.
Technical background
The organic material composting microbial inoculum mainly contains vinelandii, phosphate solubilizing bacteria, cellulose-decomposing bacterium, complex micro organism fungicide etc. both at home and abroad at present.Wherein complex microorganism becomes in recent years an important directions of exploitation both at home and abroad because its accommodative ability of environment is strong, and effect is comparatively stable, and representational product has Japanese enzymatic microorganism, EM bacterium etc.
The release one after another complex microorganism preparations of various combination of the more existing at present units of China, but general problem is in complex microorganism screening and culturing process, lack basic theoretical direction and sophisticated production technique, comparatively not familiar to the fermenting process of compound cultivation and the interaction between each flora thereof and mechanism thereof, cause the product total viable count can not keep certain level, unstable product quality has influenced its effect in use.
Summary of the invention
(1) technical problem that will solve
The objective of the invention is to disclose a kind of complex micro organism fungicide and production method thereof at above-mentioned the deficiencies in the prior art, the present invention can solve collaborative, the symbiotic relationship between the different microorganisms preferably, thereby guarantees stable action effect.
(2) technical scheme
The present invention filters out the combination of best stabilized symbiosis bacterial classification from 30 do not belong to together, plant, by single culture screening high-yield strains.The bacterial classification that filters out is yeast saccharomyces cerevisiae (Saccharomycescerevisiae), plant lactobacillus (lactobacillus plantarum), Mucor racemosus (Mucorracemosus), aspergillus oryzae (Aspergillus oryzae).
According to above-mentioned The selection result, the present invention has designed a kind of complex micro organism fungicide, comprises yeast saccharomyces cerevisiae 5.0 * 10
8～1.0 * 10
10Cfu/mL, plant lactobacillus 5.0 * 10
8～1.0 * 10
10Cfu/mL, Mucor racemosus 1.0 * 10
2～5.0 * 10
4Cfu/mL, aspergillus oryzae 1.0 * 10
3～5.0 * 10
4Cfu/mL;
Preferably, yeast saccharomyces cerevisiae 1.0 * 10
9～6.0 * 10
9Cfu/mL, plant lactobacillus 1.0 * 10
9～6.0 * 10
9Cfu/mL, Mucor racemosus 5.0 * 10
2～5.0 * 10
3Cfu/mL, aspergillus oryzae 4.0 * 10
3～1.0 * 10
4Cfu/mL.
The production method of complex micro organism fungicide of the present invention comprises the steps:
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, to cultivate 2～6 days under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, cultivated 1～3 day under 15～42 ℃ of conditions of plant lactobacillus, make actication of culture;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, 100～200r/min shaking table was cultivated 2～6 days, static cultivation is 1～3 day under 15～42 ℃ of conditions of plant lactobacillus, single bacterial classification liquid culture OD
600Stop in the time of between the value 3.0-4.0 cultivating, make first order seed;
3). secondary seed is cultivated: be 5～20% inoculum size by the volume ratio of liquid nutrient medium, be inoculated in first order seed in the fermentor tank respectively, under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, stirring velocity is 100～200r/min, air flow is 1:0.5～1.5, cultivated 2～6 days, and cultivated 1～3 day under 15～42 ℃ of conditions of plant lactobacillus, make secondary seed;
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 5～20% inoculum size, and secondary seed is inoculated in the fermentor tank, carries out high density fermentation and cultivates, and obtains microbial inoculum.
Wherein, step 1), 2), 3) in yeast saccharomyces cerevisiae, Mucor racemosus, the used substratum of aspergillus oryzae be the PDA substratum, the used substratum of plant lactobacillus is the MRS substratum.The difference of solid medium and liquid substratum is that solid medium has added an amount of agar in preparation process.
Wherein, the prescription of the substratum that step 4) is used is by mass percentage: molasses 3～20%, ammonium sulfate 0.5～10%, peptone 0.5～5%, lime carbonate 0.5～3%, surplus are water;
Described high density fermentation is cultivated can adopt fed batch cultivation (FBC) mode, and wherein feed supplement carbon source is: glucose, glycerine, sucrose, molasses are any or its mixture wherein; Nitrogenous source is: ammonium sulfate, extractum carnis, peptone be any or its mixture wherein;
And culturing process is regulated and control conditions such as fermentation pH, dissolved oxygen, mixing speed and temperature stage by stage.Specifically the fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval, remain on the aerobic conditions fermentation, air flow 1:1～1.5, regulation and control fermentation dissolved oxygen 5～15%, mixing speed 150～200r/min, 1～3 hour mixing chamber interval, stirred 25～35 ℃ of temperature 1～3 minute; B, slightly soluble oxygen and anaerobism cultivation stage: 24～96 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at interval, 3～5 hours mixing chamber intervals, stirred 3～5 minutes, using lime carbonate to regulate pH keeps being stabilized between 3.8～4.5 25～35 ℃ of temperature.
Complex micro organism fungicide of the present invention can be used for the organic materials compost, comprises the compost of animal excrement, straw, mud or the like.
(3) beneficial effect
Complex micro organism fungicide total viable count height provided by the invention acts synergistically, and effect is stable, can effectively improve the speed of becoming thoroughly decomposed of organic materials, can be used for producing biological organic fertilizer, thereby has reduced the cost of agriculture production.
Embodiment
The present invention is further illustrated below in conjunction with embodiment, but be not used for limiting invention which is intended to be protected.
The formation of embodiment 1 complex micro organism fungicide
Total count is 3.1 * 10
9Cfu/mL
Yeast saccharomyces cerevisiae 1.6 * 10
9Cfu/mL
Plant lactobacillus 1.5 * 10
9Cfu/mL
Mucor racemosus 4.1 * 10
2Cfu/mL
Aspergillus oryzae 6.2 * 10
3Cfu/mL
The formation of embodiment 2 complex micro organism fungicides
Total count is 1.0 * 10
10Cfu/mL
Yeast saccharomyces cerevisiae 1.0 * 10
10Cfu/mL
Plant lactobacillus 5.0 * 10
8Cfu/mL
Mucor racemosus 5.0 * 10
4Cfu/mL
Aspergillus oryzae 1.0 * 10
3Cfu/mL
The formation of embodiment 3 complex micro organism fungicides
Total count is 1.0 * 10
10Cfu/mL
Yeast saccharomyces cerevisiae 5.0 * 10
8Cfu/mL
Plant lactobacillus 1.0 * 10
10Cfu/mL
Mucor racemosus 1.0 * 10
2Cfu/mL
Aspergillus oryzae 5.0 * 10
4Cfu/mL
The formation of embodiment 4 complex micro organism fungicides
Total count is 6.0 * 10
9Cfu/mL
Yeast saccharomyces cerevisiae 1.0 * 10
9Cfu/mL
Plant lactobacillus 5.0 * 10
9Cfu/mL
Mucor racemosus 5.0 * 10
2Cfu/mL
Aspergillus oryzae 4.0 * 10
3Cfu/mL
The formation of embodiment 5 complex micro organism fungicides
Total count is 6.0 * 10
9Cfu/mL
Yeast saccharomyces cerevisiae 5.0 * 10
9Cfu/mL
Plant lactobacillus 1.0 * 10
9Cfu/mL
Mucor racemosus 5.0 * 10
3Cfu/mL
Aspergillus oryzae 1.0 * 10
4Cfu/mL
The production method of embodiment 6 complex micro organism fungicides
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, to cultivate 3 days under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, cultivated 2 days under 37 ℃ of conditions of plant lactobacillus;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, the 150r/min shaking table was cultivated 3 days, static cultivation is 2 days under 37 ℃ of conditions of plant lactobacillus, make first order seed, yeast saccharomyces cerevisiae, Mucor racemosus, aspergillus oryzae and each bacteria suspension optical density(OD) OD of plant lactobacillus when finishing to cultivate
600Value all reaches 4.0;
3). secondary seed is cultivated: be 10% inoculum size by the volume ratio of liquid nutrient medium, first order seed is inoculated into respectively in the fermentor tank of 100L, the cumulative volume of nutrient solution is 60L in the fermentor tank, under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, stirring velocity is 150r/min, and air flow is 1:1, cultivates 3 days, cultivated 2 days under 37 ℃ of conditions of plant lactobacillus, make secondary seed;
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 15% inoculum size, secondary seed is inoculated in 1 ton the fermentor tank, and the substratum cumulative volume in the fermentor tank is 700L, carries out high density fermentation and cultivates, and obtains microbial inoculum,
Wherein, step 1), 2), 3) in yeast saccharomyces cerevisiae, Mucor racemosus, the used substratum of aspergillus oryzae be the PDA substratum, the used substratum of plant lactobacillus is the MRS substratum.
Wherein, the prescription of the substratum that step 4) is used is by mass percentage: molasses 8%, ammonium sulfate 5%, peptone 2.5%, lime carbonate 2%, surplus are water;
High density fermentation is cultivated and is adopted the fed batch cultivation mode, and wherein feed supplement carbon source is: glucose, glycerine, sucrose, molasses; Nitrogenous source is: ammonium sulfate, extractum carnis, peptone.
The fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval remains on the aerobic conditions fermentation, air flow 1:1.2, regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, 2 hours mixing chamber intervals, stirred 30 ℃ of temperature 2 minutes; B, slightly soluble oxygen and anaerobism cultivation stage: 60 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation is stirred at interval, 4 hours mixing chamber intervals, stirred 4 minutes, use lime carbonate to regulate and stablize pH4.0,30 ℃ of temperature.
The production method of embodiment 7 complex micro organism fungicides
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, to cultivate 6 days under yeast saccharomyces cerevisiae, Mucor racemosus, 15 ℃ of conditions of aspergillus oryzae, cultivated 1 day under 42 ℃ of conditions of plant lactobacillus;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 15 ℃ of conditions of aspergillus oryzae, the 200r/min shaking table was cultivated 6 days, static cultivation is 3 days under 15 ℃ of conditions of plant lactobacillus, make first order seed, yeast saccharomyces cerevisiae, Mucor racemosus, aspergillus oryzae and each bacteria suspension optical density(OD) OD of plant lactobacillus when finishing to cultivate
600Value all reaches 3.0;
3). secondary seed is cultivated: be 5% inoculum size by the volume ratio of liquid nutrient medium, first order seed is inoculated into respectively in the fermentor tank of 100L, the cumulative volume of nutrient solution is 60L in the fermentor tank, under yeast saccharomyces cerevisiae, Mucor racemosus, 15 ℃ of conditions of aspergillus oryzae, stirring velocity is 200r/min, and air flow is 1:0.5, cultivates 6 days, cultivated 1 day under 42 ℃ of conditions of plant lactobacillus, make secondary seed.
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 20% inoculum size, secondary seed is inoculated in 1 ton the fermentor tank, and the substratum cumulative volume in the fermentor tank is 700L, carries out high density fermentation and cultivates, and obtains microbial inoculum.
Wherein, step 1), 2), 3) in yeast saccharomyces cerevisiae, Mucor racemosus, the used substratum of aspergillus oryzae be the PDA substratum, the used substratum of plant lactobacillus is the MRS substratum.
Wherein, the prescription of the substratum that step 4) is used is by mass percentage: molasses 3%, ammonium sulfate 10%, peptone 5%, lime carbonate 0.5%, surplus are water;
High density fermentation is cultivated and is adopted the fed batch cultivation mode, and wherein feed supplement carbon source is: glucose, glycerine; Nitrogenous source is: extractum carnis, peptone.
The fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval remains on the aerobic conditions fermentation, air flow 1:1, regulation and control fermentation dissolved oxygen 5%, mixing speed 200r/min, 3 hours mixing chamber intervals, stirred 35 ℃ of temperature 3 minutes; B, slightly soluble oxygen and anaerobism cultivation stage: 96 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation is stirred at interval, 3 hours mixing chamber intervals, stirred 3 minutes, use lime carbonate to regulate and stablize pH4.5,35 ℃ of temperature.
The production method of embodiment 8 complex micro organism fungicides
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, to cultivate 2 days under yeast saccharomyces cerevisiae, Mucor racemosus, 38 ℃ of conditions of aspergillus oryzae, cultivated 3 days under 15 ℃ of conditions of plant lactobacillus;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 38 ℃ of conditions of aspergillus oryzae, the 100r/min shaking table was cultivated 2 days, static cultivation is 1 day under 42 ℃ of conditions of plant lactobacillus, make first order seed, yeast saccharomyces cerevisiae, Mucor racemosus, aspergillus oryzae and each bacteria suspension optical density(OD) OD of plant lactobacillus when finishing to cultivate
600Value all reaches 3.0;
3). secondary seed is cultivated: be 20% inoculum size by the volume ratio of liquid nutrient medium, first order seed is inoculated into respectively in the fermentor tank of 100L, the cumulative volume of nutrient solution is 60L in the fermentor tank, under yeast saccharomyces cerevisiae, Mucor racemosus, 38 ℃ of conditions of aspergillus oryzae, stirring velocity is 100r/min, and air flow is 1:1.5, cultivates 2 days, cultivated 3 days under 15 ℃ of conditions of plant lactobacillus, make secondary seed.
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 5% inoculum size, secondary seed is inoculated in 1 ton the fermentor tank, and the substratum cumulative volume in the fermentor tank is 700L, carries out high density fermentation and cultivates, and obtains microbial inoculum.
Wherein, step 1), 2), 3) in yeast saccharomyces cerevisiae, Mucor racemosus, the used substratum of aspergillus oryzae be the PDA substratum, the used substratum of plant lactobacillus is the MRS substratum.
Wherein, the prescription of the substratum that step 4) is used is by mass percentage: molasses 20%, ammonium sulfate 0.5%, peptone 0.5%, lime carbonate 3%, surplus are water;
High density fermentation is cultivated and is adopted the fed batch cultivation mode, and wherein feed supplement carbon source is: sucrose, molasses; Nitrogenous source is: ammonium sulfate, extractum carnis.
The fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval remains on the aerobic conditions fermentation, air flow 1:1.5, regulation and control fermentation dissolved oxygen 15%, mixing speed 150r/min, 1 hour mixing chamber interval, stirred 25 ℃ of temperature 1 minute; B, slightly soluble oxygen and anaerobism cultivation stage: 24 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation is stirred at interval, 5 hours mixing chamber intervals, stirred 5 minutes, use lime carbonate to regulate and stablize pH3.8,25 ℃ of temperature.
Embodiment 9 is the compost of main raw material with the chicken manure
Form: chicken manure 80% wheat straw 18% wheat bran 1.7% brown sugar 0.1% complex micro organism fungicide 0.2% of the present invention
Making processes:
Must add this microbial inoculum again and fully stir evenly earlier with the brown sugar water dissolution during batching, then even the dialling of diluent is sprinkled upon on the starting material, and fully stir (best with stirrer) is even.
Material moisture generally should be controlled at 50～60%, and the C/N ratio is adjusted to 25～30:1, and the pH value is neutrality or weakly alkaline, and material particular diameter is between 2～60mm.During fermentation, bank is chevron or trapezoidal the fermentation into strips.As pile the strip chevron, and the heap height generally is no more than 80cm, the wide about 120～150cm in bottom, and length is not limit; As pile trapezoidal, the heap height generally be no more than 50cm, length and width are not limit.(the C/N here refers to total carbon and the total nitrogen ratio in the middle of the material.The C/N value is high, contains the high material of N amount by interpolation and regulates, and the C/N value is low, regulates by adding the high material of C content.)
In the fermenting process, should observe temperature of charge and change, after temperature rises to more than 60 ℃, material turning in per 2～3 days once, turning is 3～4 times continuously.Ferment and finished substantially in 7～10 days.The compost maturity effect: as shown in table 1, adding this microbial inoculum has certain facilitation effect for the decomposition of organic carbon.
The variation of organic carbon after this microbial inoculum of table 1 interpolation
| Raw material becomes thoroughly decomposed | Starting material organic carbon content % | Become thoroughly decomposed and finish material organic carbon content % | Material organic carbon content decline % |
| Chicken manure+wheat straw+0.2% microbial inoculum | 42.63 | 26.48 | 37.88 |
| Chicken manure+wheat straw | 42.63 | 30.36 | 28.78 |
Case study on implementation 10 is the compost of main raw material with the cow dung
Form: cow dung 95% wheat bran 4.7% brown sugar 0.1% complex micro organism fungicide 0.2% making processes of the present invention: earlier that a certain amount of brown sugar is water-soluble, the amount of brown sugar is 1 ‰ of a composting material dry weight, add this microbial inoculum of 2 ‰, with the sorbing material of a spot of wheat bran as microbial inoculum.With wheat bran and composting material mixing, carry out compost then, and fully stir (best with stirrer) is even.
Material moisture generally should be controlled at 50～60%, and the C/N ratio is adjusted to 25～30:1, and the pH value is neutrality or weakly alkaline, and material particular diameter is between 2～60mm.
During fermentation, the heap height generally is no more than 80cm, the wide 120～150cm in bottom, and length is not limit; As pile trapezoidal, the heap height generally be no more than 50cm, length and width are not limit.After bank is good, can cover plastic cloth, and with suitably shading such as straw screen or mat.
In the fermenting process, should observe temperature of charge and change, after temperature rises to more than 60 ℃, turning in per 2～3 days once, turning is 3～4 times continuously.Ferment and finished substantially in 7～10 days.
The compost maturity effect: this microbial inoculum of adding as shown in table 2 has certain facilitation effect for the decomposition of organic carbon.
The variation of organic carbon after this microbial inoculum of table 2 interpolation
| Raw material becomes thoroughly decomposed | Starting material organic carbon content % | Become thoroughly decomposed and finish material organic carbon content % | Material organic carbon content decline % |
| Cow dung+wheat straw+2 ‰ microbial inoculum | 35.67 | 27.48 | 22.96 |
| Cow dung+wheat straw | 35.67 | 28.87 | 19.06 |
Case study on implementation 11 is the compost of main raw material with the stalk
Form: maize straw 98% complex micro organism fungicide 0.2% of the present invention, brown sugar 1.8%
Making processes: must earlier the brown sugar water be dissolved (separating) during batching, add this microbial inoculum again and fully stir evenly, then even the dialling of diluent is sprinkled upon on the starting material.The maize straw that adds this microbial inoculum is placed into the soil.
The result: behind the maize straw, in the first two months of surviving the winter (to December 5), maize straw adds this microbial inoculum of spray, and an average day decomposition amount reaches 1.08 grams, and not adding the average day decomposition amount of this microbial inoculum of spray is 0.38 gram.This explanation sprays the decomposition that this microbial inoculum can quicken stalk.
Application case 12 is the compost of main raw material with mud
Form: mud: corn cob: returned sluge: flyash=14:2:3:1 (weight ratio), complex micro organism fungicide 0.2% of the present invention, brown sugar 0.1%.
Making processes: manually with mixing of materials well after, drop into fermentation vat.Material moisture is controlled at 55～65%, and the suitableeest 60%.The compost volume is 12m
3Same recipe material preparation two heaps, the initial C/N of composting material is 25.
Earlier that a certain amount of brown sugar is water-soluble, the amount of brown sugar is 1 ‰ of a composting material dry weight, adds this microbial inoculum of 2 ‰, with the sorbing material of a spot of wheat bran as microbial inoculum.With wheat bran and composting material mixing, carry out compost then, and fully stir evenly with stirrer.Compost growing * wide * highly carrying out in the cement pit of 30m * 6m * 1m, mechanical heap turning over device is arranged at fermentation vat top, and the bottom is equipped with the PVC ventpipe.Regularly ventilate every day, fermented altogether 48 days.
The result of becoming thoroughly decomposed: the initial C/N of composting material is 25, is reduced to C/N to be the standard of becoming thoroughly decomposed below 20, and the processing of inoculating this microbial inoculum reached in advance than the processing of not inoculating this microbial inoculum in 6 days becomes thoroughly decomposed.
Claims (6)
1, a kind of complex micro organism fungicide that is used for compost fermentation is characterized in that this microbial inoculum is a yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) 5.0 * 10
8～1.0 * 10
10Cfu/mL, plant lactobacillus (lactobacillus plantarum) 5.0 * 10
8～1.0 * 10
10Cfu/mL, Mucor racemosus (Mucor racemosus) 1.0 * 10
2～5.0 * 10
4Cfu/mL and aspergillus oryzae (Aspergillusoryzae) 1.0 * 10
3～5.0 * 10
4The composite bacteria of cfu/mL.
2, complex micro organism fungicide according to claim 1 is characterized in that this microbial inoculum is a yeast saccharomyces cerevisiae 1.0 * 10
9～6.0 * 10
9Cfu/mL, plant lactobacillus 1.0 * 10
9～6.0 * 10
9Cfu/mL, Mucor racemosus 5.0 * 10
2～5.0 * 10
3Cfu/mL and aspergillus oryzae 4.0 * 10
3～1.0 * 10
4The composite bacteria of cfu/mL.
3, complex micro organism fungicide according to claim 1 is characterized in that this microbial inoculum is a yeast saccharomyces cerevisiae 1.6 * 10
9Cfu/mL, plant lactobacillus 1.5 * 10
9Cfu/mL, Mucor racemosus 4.1 * 10
2Cfu/mL and aspergillus oryzae 6.2 * 10
3The mixture of cfu/mL.
4, a kind of production method as each described complex micro organism fungicide of claim 1～3 is characterized in that comprising following steps:
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, with cultivating 2～6 days under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, cultivated 1～3 day under 15～42 ℃ of conditions of plant lactobacillus;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, 100～200r/min shaking table was cultivated 2～6 days, static cultivation is 1～3 day under 15～42 ℃ of conditions of plant lactobacillus, single bacterial classification liquid culture OD
600Stop in the time of between the value 3.0-4.0 cultivating, make first order seed;
3). secondary seed is cultivated: be 5～20% inoculum size by the volume ratio of liquid nutrient medium, be inoculated in first order seed in the fermentor tank respectively, under yeast saccharomyces cerevisiae, Mucor racemosus, 15～38 ℃ of conditions of aspergillus oryzae, stirring velocity is 100～200r/min, air flow is 1:0.5～1.5, cultivated 2～6 days, and cultivated 1～3 day under 15～42 ℃ of conditions of plant lactobacillus, make secondary seed;
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 5～20% inoculum size, and secondary seed is inoculated in the fermentor tank, carries out high density fermentation and cultivates, and obtains microbial inoculum,
Wherein, described high density fermentation is cultivated and adopted the fed batch cultivation mode, and wherein feed supplement carbon source is: glucose, glycerine, sucrose, molasses are any or its mixture wherein; Nitrogenous source is: ammonium sulfate, extractum carnis, peptone be any or its mixture wherein,
The fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval, remain on the aerobic conditions fermentation, air flow 1:1～1.5, regulation and control fermentation dissolved oxygen 5～15%, mixing speed 150-200r/min, 1～3 hour mixing chamber interval, stirred 25～35 ℃ of temperature 1～3 minute; B, slightly soluble oxygen and anaerobism cultivation stage: 24～96 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation, stir at interval, 3～5 hours mixing chamber intervals, stirred 3～5 minutes, using lime carbonate to regulate pH keeps being stabilized between 3.8～4.5 25～35 ℃ of temperature.
5, a kind of production method of complex micro organism fungicide as claimed in claim 4 is characterized in that:
1). slant culture: will be inoculated on the solid medium respectively under yeast saccharomyces cerevisiae, plant lactobacillus, Mucor racemosus, the aspergillus oryzae original strain aseptic condition, to cultivate 3 days under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, cultivated 2 days under 37 ℃ of conditions of plant lactobacillus;
2). first order seed is cultivated: be inoculated in liquid nutrient medium respectively under the bacterial classification aseptic condition with the step 1) cultivation, under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, the 150r/min shaking table was cultivated 3 days, and static cultivation is 2 days under 37 ℃ of conditions of plant lactobacillus, single bacterial classification liquid culture OD
600Stop in the time of between the value 3.0-4.0 cultivating, make first order seed;
3). secondary seed is cultivated: be 10% inoculum size by the volume ratio of liquid nutrient medium, be inoculated in first order seed in the fermentor tank respectively, under yeast saccharomyces cerevisiae, Mucor racemosus, 30 ℃ of conditions of aspergillus oryzae, stirring velocity is 150r/min, air flow is 1:1, cultivated 3 days, and cultivated 2 days under 37 ℃ of conditions of plant lactobacillus, make secondary seed;
4). mixing fermentation culture: by the volume ratio of liquid nutrient medium is 15% inoculum size, and secondary seed is inoculated in the fermentor tank, carries out high density fermentation and cultivates, and obtains microbial inoculum,
Wherein said high density fermentation is cultivated the fed batch cultivation mode that adopts, and wherein feed supplement carbon source is: molasses; Nitrogenous source is: peptone,
The fermentation culture process comprises: a, aerobic cultivation stage: in initial 0～24 hour, ventilation at interval remains on the aerobic conditions fermentation, air flow 1:1.2, regulation and control fermentation dissolved oxygen 10%, mixing speed 180r/min, 2 hours mixing chamber intervals, stirred 30 ℃ of temperature 2 minutes; B, slightly soluble oxygen and anaerobism cultivation stage: 60 hours, keep fermented liquid upper strata slightly soluble oxygen condition, static cultivation is stirred at interval, 4 hours mixing chamber intervals, stirred 4 minutes, use lime carbonate to regulate pH and keep being stabilized in 4.0,30 ℃ of temperature.
6, a kind of as the purposes of each described complex micro organism fungicide of claim 1～3 in the organic materials compost.
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| CN101416610B (en) * | 2008-06-24 | 2011-08-17 | 李季 | Pigpen fermentation bed and use thereof |
| CN102050646A (en) * | 2009-11-10 | 2011-05-11 | 江苏省农业科学院 | In-situ composting method and device of crop straws |
| CN102010268A (en) * | 2010-10-21 | 2011-04-13 | 哈尔滨工业大学 | Compost activating agent and method for compositing organic solid wastes by using same |
| CN102174398B (en) * | 2010-12-14 | 2012-10-31 | 北京沃土天地生物科技有限公司 | Composite microbiological bacterial agent used for returning maize straws to field and preparation method and applications thereof |
| CN102212494B (en) * | 2011-04-12 | 2013-03-13 | 王颖 | Organic matter decomposing inoculant, and preparation method and application thereof |
| CN103172420A (en) * | 2012-11-08 | 2013-06-26 | 曾凡强 | Broad-spectrum applicable organic material decomposition agent |
| CN103172421A (en) * | 2013-02-21 | 2013-06-26 | 北京沃土天地生物科技有限公司 | Treatment method of fruit/vegetable waste |
| CN103466910A (en) * | 2013-08-23 | 2013-12-25 | 福州大用生物应用科技有限公司 | Compound micro ecological bacteria agent used for live pig excrement treatment |
| CN103444784B (en) * | 2013-08-23 | 2015-09-09 | 福州大用生物应用科技有限公司 | A kind of compound micro-ecological preparation for live pig auxiliary epidemic prevention |
| CN103992153B (en) * | 2014-06-06 | 2016-04-20 | 武汉市蔬菜科学研究所 | The biological organic fertilizer that a kind of early spring, booth was suitable for and preparation method |
| CN107365215A (en) * | 2016-05-11 | 2017-11-21 | 沈阳市现代三川生物技术研究所 | One kind contains diatomaceous microbe soil insecticide preparation method |
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