CN107879774A - A kind of preparation method of biological decomposing agent - Google Patents
A kind of preparation method of biological decomposing agent Download PDFInfo
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- CN107879774A CN107879774A CN201711090692.3A CN201711090692A CN107879774A CN 107879774 A CN107879774 A CN 107879774A CN 201711090692 A CN201711090692 A CN 201711090692A CN 107879774 A CN107879774 A CN 107879774A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- 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
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- 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
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Abstract
The invention discloses a kind of preparation method of biological decomposing agent, belong to agriculture field.The present invention obtains acc deaminase positive strain by screening plant nitrogen-fixing bacteria in soil, being manured into soil can not only promote biology decomposed, there is fixation to the nitrogen in soil simultaneously, directly act on plant root, plant-growth promoting rhizobacteria has acc deaminase activity, ACC can be decomposed ammonification and α-batanone acid reduces ethylene synthase, so as to reduce sensitiveness of the plant to adverse circumstance, plant stress-resistance ability is improved, and the phytoremediation of Organic Pollution and heavy-metal contaminated soil can be promoted.The present invention solves the problems, such as to produce with foul gas in current organic material decomposing agent digest process and decomposed efficiency is low.
Description
Technical field
The invention belongs to agriculture field, and in particular to a kind of preparation method of biological decomposing agent.
Background technology
Decomposing agent refers to that various organic materials can be accelerated（It is dirty including agricultural crop straw, feces of livestock and poultry, house refuse and city
Mud etc.）Decompose, decomposed living microorganisms preparation.With the high speed development of China's agricultural and processing industry, China has become generation
The maximum country of agriculture organic materials yield in boundary, it is annual caused by agriculture organic materials in terms of tens tons, wherein, every year
Produce nearly 4,000,000,000 tons of feces of livestock and poultry, 700,000,000 tons of agricultural crop straw.In recent years, domestic and international agricultural wastes application technology as the second resource
Research achieves larger development, and agricultural wastes recycling gradually steps into the scientific new stage.In all utilization sides
In formula, the method that high temperature aerobic composting is a kind of currently used effectively processing organic waste is carried out using microbial technique,
Because nutritive loss compared with it is small, innoxious degree is high, treating capacity is big, cost is relatively low, has turned into suitable for many advantages, such as factorial praluction
The preferred processing mode of feces of livestock and poultry and agricultural crop straw.
Traditional compost fermentation is to complete organic substance biotransformation using the indigenous microorganism in raw material.Compost
Fermentation process is broadly divided into temperature raising period, megathermal period, cooldown period three phases, and there is its unique microbial population in each stage.
Temperature raising period, mesophilic property microorganism are released by the use of soluble and readily degradable organic matter as nutrition and energy source, rapid propagation
Heat energy is released, temperature is constantly increased.Megathermal period, mesophilic property microorganism are suppressed, and thermophilic microorganism gradually replaces,
Active microorganism is mainly thermophilic fungi and actinomyces etc..As organic matter consumes or converts, temperature declines, mesophilic micro- life
Thing starts to turn into dominant population again.Therefore, according to composting process microorganism diversity feature, at multiple-microorganism assistance
Reason composting material is to accelerate decomposed, the key of raising compost quality of finished.Carried out using organic matter decomposing inoculant it is decomposed, it is not only right
Organic materials has powerful decomposed effect, and also breeds a large amount of function bacteriums during the fermentation and produce a variety of special efficacy metabolism productions
Thing（Such as hormone, antibiotic）, so as to stimulate crop growth, improve crop disease-resistant, drought resisting, cold tolerance.Function bacterium
, can fixed nitrogen, phosphorus decomposing, potassium decomposing, increase soil nutrient, improvement soil texture, raising chemical fertilizer utilization ratio into after soil.Organic materials
Decomposing agent is safe to use, can handle gas chromatography material, nontoxic, harmless, pollution-free.
At present organic matter decomposing inoculant mainly by can in fermentation or growth course eccrine fiber element enzyme and protease it is true
The microorganisms such as bacterium, bacterium and actinomyces are combined, and are by mixed fermentation or after single fermentation is simply mixed later
A kind of organic matter decomposing inoculant.But because the microbe species of compounding are different, characteristic, function and the difference on effect of its product are very
Greatly.Produced using organic matter decomposing inoculant during organic fertilizer often along with the generation of foul gas, make one poor appetite,
Feel dizzy, Nausea and vomiting, and decomposed effect is undesirable.Therefore, produce that a kind of decomposed effect is good and eliminate peculiar smell in use
Decomposing agent exist the very big market demand.
The content of the invention
The technical problems to be solved by the invention：For being produced in current organic material decomposing agent digest process with foul gas
A kind of the problem of raw and decomposed efficiency is low, there is provided preparation method of biological decomposing agent.
In order to solve the above technical problems, the present invention is using technical scheme as described below：
A kind of preparation method of biological decomposing agent, it is characterised in that the preparation method comprises the following steps：
（1）Soil sampling is well mixed with sterilized water, shaking table vibration, obtains soil sample mixed liquor, and soil sample mixed liquor is added into fixed nitrogen bacterium solution
Cultivated in body culture medium, fixed nitrogen bacteria culture fluid must be enriched with, repeat enrichment culture 3 times, obtain multiple culture enrichment fixed nitrogen bacteria culture fluid, will
The normal saline dilution that multiple culture enrichment fixed nitrogen bacteria culture fluid mass fraction is 0.9% is to 10-5Dilute level, fixed nitrogen after must diluting
Bacterium solution, fixed nitrogen bacterium solution is coated on nitrogen-fixing bacteria separation plating medium after taking dilution, culture, obtains nitrogen-fixing bacteria bacterium colony, repeats line training
Support 2 ~ 3 times, nitrogen-fixing bacteria bacterium colony must be purified；
（2）Picking purifying nitrogen-fixing bacteria colony inoculation is cultivated into nitrogen-fixing bacteria fluid nutrient medium, fixed nitrogen bacteria culture fluid is obtained, by nitrogen-fixing bacteria
Nutrient solution is seeded to DF culture medium shaken cultivations, obtains nutrient solution, and nutrient solution is seeded in ADF culture mediums and cultivated, repeated inoculation
Culture 2 ~ 3 times, obtains deaminase positive bacteria culture fluid, centrifuges, takes precipitation, dries, obtains deaminase positive bacteria；
（3）Sludge is taken to be mixed with sterilized water, culture, sludge suspension is obtained, is stood, takes the supernatant of sludge suspension, is inoculated with de-
Enrichment culture in sulphur bacterium enriched medium, obtains enrichment culture liquid, and enrichment culture liquid is diluted into 10-5Dilution level, after obtaining dilution
Desulfurization bacterium bacterium solution, desulfurization bacterium bacterium solution after dilution is seeded to desulfurization bacterium screening and culturing medium culture, obtains desulfurization bacterium bacterium colony, picking bacterium footpath
Maximum desulfurization bacterium bacterium colony streak inoculation repeats line culture 2 ~ 3 times, obtains purifying desulfurization to desulfurization bacterium screening and culturing medium culture
Bacterium；
（4）EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium are well mixed, obtained
Decomposing microbial inoculum is mixed, mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran are well mixed, obtain biological decomposing agent.
The step（1）Middle pedotheque is derived from the potato rhizosphere soil in growth period；Nitrogen-fixing bacteria fluid nutrient medium
It is formulated according to the mass fraction, to take 10 ~ 15 parts of sucrose, K2HPO4·3H20.2 ~ 0.5 part of O, NaCl0.2 ~ 0.5 part, CaCO31~2
Part, MgSO4·7H20.2 ~ 0.4 part of O, 0.5 ~ 0.8 part of yeast extract, 1000 parts of distilled water, pH 7. 0 ± 0.2；Nitrogen-fixing bacteria separate
Plating medium：Add 20 ~ 25 parts of agar into nitrogen-fixing bacteria fluid nutrient medium, other components are constant.
The step（1）The mass ratio of middle soil sample and sterilized water is 1：9；Soil sample mixed liquor and nitrogen-fixing bacteria fluid nutrient medium
Volume ratio is 1：8, the condition of culture of enrichment fixed nitrogen bacteria culture fluid is 25 ~ 28 DEG C, 100r/min shaking tables shaken cultivation 2 ~ 3 days；It is pure
The condition of culture for changing nitrogen-fixing bacteria bacterium colony is 25 ~ 29 DEG C of 2 ~ 3d of culture.
The step（2）The formula of middle DF culture mediums is according to the mass fraction, to take KH2PO44 ~ 6 parts, Na2HPO46 ~ 10 parts,
MgSO4·7H20.2 ~ 0.5 part of O, 2 ~ 6 parts of glucose, 3 ~ 5 parts of sodium gluconate, 1 ~ 2 part of citric acid, (NH4) 2SO42 ~ 4 parts,
0.1 ~ 0.2 part of one solution of component, 0.1 ~ 0.2 part of two solution of component, 1000 parts of water, pH 7. 2 ± 0.2, wherein component one be by
Mass fraction meter, takes H3BO30.01 ~ 0.03 part, MnSO4·H20.01 ~ 0.02 part of O, ZnSO4·7H20.1 ~ 0.3 part of O,
CuSO4·5H20.07 ~ 0.09 part of O, MoO30.01 ~ 0.02 part, 100 parts of water；Component two is by FeSO47H2O and aseptic distillation
Water in mass ratio 1：100 is well mixed, produces；The formula of ADF culture mediums is by isolated purifying nitrogen-fixing bacteria in mass ratio 1：
30 are seeded to and are free of (NH4 ) 2SO4In the sterilizing DF culture mediums of component, ADF culture mediums are obtained, pH is 7 ~ 7. 5.
The step（2）The mass ratio of middle purifying nitrogen-fixing bacteria bacterium colony and nitrogen-fixing bacteria fluid nutrient medium is 1：5, condition of culture is
25 ~ 30 DEG C, 24 ~ 36h of 200r/min shaken cultivations；The volume ratio of fixed nitrogen bacteria culture fluid and DF culture mediums is 1：5；Nutrient solution presses body
Product ratio 1：50 are seeded in ADF culture mediums 24 ~ 48h of shaken cultivation under the conditions of 25 ~ 30 DEG C.
The step（3）Middle desulfurization bacterium screening sample is derived from the sludge of sewage deposited bottom；Desulfurization bacterium enriched medium
It is formulated according to the mass fraction, to take 10 ~ 12 parts of peptone, 5 ~ 7 parts of beef extract, 5 ~ 8 parts of sodium chloride, 1000 parts of water, pH is 7.0 ±
0.2；The formula of desulfurization bacterium screening and culturing medium is according to the mass fraction, to take KH2PO42 ~ 5 parts, 15 ~ 18 parts of peptone, beef extract 3 ~ 5
Part, Na2HPO4·12H202 ~ 5 part, 3 ~ 5 parts of sodium chloride, 20 ~ 25 parts of agar, 1000 parts of water, pH is 7.0 ± 0.2.
The step（3）The mass ratio of middle sludge and sterilized water is 1：10, condition of culture is 25 ~ 30 DEG C, 120r/min shakes
Swing 3 ~ 5h of culture；The mass ratio of sludge suspension supernatant and desulfurization bacterium enriched medium is 1：10, condition of culture is 25 ~ 30
DEG C, 120r/min enrichment cultures 2 ~ 3 days.
The step（4）Middle mixing decomposing microbial inoculum is EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, deamination enzyme positive
Bacterium, purifying desulfurization bacterium in mass ratio 5：2：1：4：4 is well mixed, and biological decomposing agent is mixing decomposing microbial inoculum, chicken manure, rice husk, rice
Chaff in mass ratio 5：7：8：4 is well mixed.
Compared with other method, advantageous effects are the present invention：
（1）The present invention obtains deaminase positive strain by screening plant nitrogen-fixing bacteria in soil, and being manured into soil can not only promote
Biology is decomposed, while has fixation to the nitrogen in soil, directly acts on plant root, plant-growth promoting rhizobacteria has deaminase
Activity, can decompose ammonification and α-batanone acid reduces ethylene synthase, so as to reduce sensitiveness of the plant to adverse circumstance, improve Genes For Plant Tolerance
Inverse ability, and the phytoremediation of Organic Pollution and heavy-metal contaminated soil can be promoted；
（2）The present invention by sludge screen obtain high-efficiency desulfurization bacterium, in digest process can to caused foul gas desulfurization,
Decomposed generation pernicious gas is avoided, pollutes environment；
（3）The present invention breeds substantial amounts of function bacterium, and produce a variety of sovereign remedies by the compounding of composite bacteria agent in digest process
Matter, decomposed, raising efficiency can be accelerated, while growth and development of plants can be stimulated, function bacterium enters soil, can fixing nitrogen, dissolving phosphor and dissolving
Potassium, increase soil nutrient, improve soil texture, and it is safe to use, it is nontoxic pollution-free.
Embodiment
Pedotheque：It is derived from the potato rhizosphere soil in growth period.
Desulfurization bacterium screening sample：It is derived from the sludge of sewage deposited bottom.
Decomposing agent base starting material：Wood chip, rice husk, chicken manure, rice bran.
Decomposed fermented bacterium：EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte.
Desulphurization Strains screening sample：Activated sludge and anaerobic sludge selected from sewage treatment plant.
Nitrogen-fixing bacteria fluid nutrient medium：According to the mass fraction, 10 ~ 15 parts of sucrose, K are taken2HPO4·3H2O0.2 ~ 0.5 part,
NaCl0.2 ~ 0.5 part, CaCO31 ~ 2 part, MgSO4·7H2O0.2 ~ 0.4 part, 0.5 ~ 0.8 part of yeast extract, 1000 parts of distilled water, pH
7.0±0.2。
Nitrogen-fixing bacteria separate plating medium：Add 20 ~ 25 parts of agar into nitrogen-fixing bacteria fluid nutrient medium, other components are constant.
DF culture mediums：According to the mass fraction, KH is taken2PO44 ~ 6 parts, Na2HPO46 ~ 10 parts, MgSO4·7H2O 0.2~0.5
Part, 2 ~ 6 parts of glucose, 3 ~ 5 parts of sodium gluconate, 1 ~ 2 part of citric acid, (NH4)2SO42 ~ 4 parts, 0.1 ~ 0.2 part of one solution of component,
0.1 ~ 0.2 part of two solution of component, 1000 parts of water, pH7. 2 ± 0.2.Wherein component one is according to the mass fraction, to take H3BO30.01~
0.03 part, MnSO4·H2O0.01 ~ 0.02 part, ZnSO4·7H2O0.1 ~ 0.3 part, CuSO4·5H2O0.07 ~ 0.09 part,
MoO30.01 ~ 0.02 part, 100 parts of water；Component two is by FeSO47H2O and sterile purified water in mass ratio 1：100 is well mixed,
Produce.
ADF culture mediums：By isolated purifying nitrogen-fixing bacteria in mass ratio 1：30 are seeded to and are free of (NH4 )2SO4Component is gone out
In bacterium DF culture mediums, ADF culture mediums are obtained, pH is 7 ~ 7. 5.
Desulfurization bacterium enriched medium：According to the mass fraction, 10 ~ 12 parts of peptone, 5 ~ 7 parts of beef extract, sodium chloride 5 ~ 8 are taken
Part, 1000 parts of water, pH is 7.0 ± 0.2.
Desulfurization bacterium screening and culturing medium：According to the mass fraction, KH is taken2PO42 ~ 5 parts, 15 ~ 18 parts of peptone, 3 ~ 5 parts of beef extract,
Na2HPO4·12H2O2 ~ 5 part, 3 ~ 5 parts of sodium chloride, 20 ~ 25 parts of agar, 1000 parts of water, pH are 7.0 ± 0.2.
A kind of preparation method of biological decomposing agent, comprises the following steps：
（1）Soil sampling in mass ratio 1：9 are well mixed with sterilized water, shaking table vibration 20min, obtain soil sample mixed liquor, soil sample is mixed
Close liquid by volume 1：8 add in nitrogen-fixing bacteria fluid nutrient mediums, in 25 ~ 28 DEG C, 100r/min shaking tables shaken cultivation 2 ~ 3 days, obtain
Fixed nitrogen bacteria culture fluid is enriched with, repeats enrichment culture 3 times, obtains multiple culture enrichment fixed nitrogen bacteria culture fluid, multiple culture is enriched with nitrogen-fixing bacteria
The normal saline dilution that nutrient solution mass fraction is 0.9% is to 10-5Level is diluted, fixed nitrogen bacterium solution after must diluting, takes fixed nitrogen after dilution
Bacterium solution is coated on nitrogen-fixing bacteria separation plating medium, 25 ~ 29 DEG C of 2 ~ 3d of culture, obtains nitrogen-fixing bacteria bacterium colony, repeats line culture 2 ~ 3
It is secondary, nitrogen-fixing bacteria bacterium colony must be purified；
（2）Picking purifying nitrogen-fixing bacteria bacterium colony in mass ratio 1：5 are seeded in nitrogen-fixing bacteria fluid nutrient medium, 25 ~ 30 DEG C, 200r/min
24 ~ 36h of shaken cultivation, fixed nitrogen bacteria culture fluid is obtained, by fixed nitrogen bacteria culture fluid by volume 1：5 are seeded to DF culture medium shaken cultivations
18 ~ 24h, nutrient solution is obtained, by nutrient solution by volume 1：50 are seeded in ADF culture mediums the shaken cultivation under the conditions of 25 ~ 30 DEG C
24 ~ 48h, repeated inoculation culture 2 ~ 3 times, deaminase positive bacteria culture fluid is obtained, centrifuge, take precipitation, dried, obtain deamination enzyme positive
Bacterium；
（3）Take sludge in mass ratio 1：10 mix with sterilized water, 25 ~ 30 DEG C, 120r/min 3 ~ 5h of shaken cultivation, obtain sludge suspension
Liquid, 2 ~ 4h is stood, takes sludge suspension supernatant in mass ratio 1：In 10 inoculation desulfurization bacterium enriched mediums, 25 ~ 30 DEG C,
120r/min enrichment cultures 2 ~ 3 days, obtain enrichment culture liquid, and enrichment culture liquid is diluted into 10-5Dilution level, desulfurization after must diluting
Bacterium bacterium solution, by desulfurization bacterium bacterium solution in mass ratio 1 after dilution：10 are seeded to desulfurization bacterium screening and culturing medium, and 25 ~ 30 DEG C are cultivated 2 ~ 3 days,
Obtain desulfurization bacterium bacterium colony, the maximum desulfurization bacterium bacterium colony streak inoculation in picking bacterium footpath to 25 ~ 30 DEG C of cultures 2 ~ 3 of desulfurization bacterium screening and culturing medium
My god, line culture 2 ~ 3 times is repeated, obtains purifying desulfurization bacterium；
（4）By EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium in mass ratio 5：
2：1：4：4 is well mixed, obtains mixing decomposing microbial inoculum, by mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran in mass ratio 5：7：8：4 is mixed
Close uniformly, obtain biological decomposing agent.
Example 1
Pedotheque：It is derived from the field-crop rhizosphere in growth period.
Desulfurization bacterium screening sample：It is derived from the sludge of sewage deposited bottom.
Decomposing agent base starting material：Wood chip, rice husk, chicken manure, rice bran.
Decomposed fermented bacterium：EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte.
Desulphurization Strains screening sample：Activated sludge and anaerobic sludge selected from sewage treatment plant.
Nitrogen-fixing bacteria fluid nutrient medium：According to the mass fraction, 10 parts of sucrose, K are taken2HPO4·3H2O0.2 parts, NaCl0.2 parts,
CaCO31 part, MgSO4·7H20.2 part of O, 0.5 part of yeast extract, 1000 parts of distilled water, pH 6.8.
Nitrogen-fixing bacteria separate plating medium：Add 20 parts of agar into nitrogen-fixing bacteria fluid nutrient medium, other components are constant.
DF culture mediums：According to the mass fraction, KH is taken2PO44 parts, Na2HPO46 parts, MgSO4·7H2O0.2 parts, glucose 2
Part, 3 parts of sodium gluconate, 1 part of citric acid, (NH4)2SO42 parts, 0.1 part of one solution of component, 0.1 part of two solution of component, water 1000
Part, pH 7.0.Wherein component one is according to the mass fraction, to take H3BO30.01 part, MnSO4·H2O0.01 parts, ZnSO4·
7H2O0.1 parts, CuSO4·5H2O0.07 parts, MoO30.01 part, 100 parts of water；Component two is by FeSO47H2O and sterile purified water
In mass ratio 1：100 is well mixed, produces.
ADF culture mediums：By isolated purifying nitrogen-fixing bacteria in mass ratio 1：30 are seeded to and are free of (NH4 )2SO4Component is gone out
In bacterium DF culture mediums, ADF culture mediums, pH 7 are obtained.
Desulfurization bacterium enriched medium：According to the mass fraction, 10 parts of peptone, 5 parts of beef extract, 5 parts of sodium chloride, water 1000 are taken
Part, pH 6.8.
Desulfurization bacterium screening and culturing medium：According to the mass fraction, KH is taken2PO42 parts, 15 parts of peptone, 3 parts of beef extract,
Na2HPO4·12H2O2 parts, 3 parts of sodium chloride, 20 parts of agar, 1000 parts of water, pH 6.8.
A kind of preparation method of biological decomposing agent, comprises the following steps：
（1）Soil sampling in mass ratio 1：9 are well mixed with sterilized water, shaking table vibration 20min, obtain soil sample mixed liquor, soil sample is mixed
Close liquid by volume 1：8 add in nitrogen-fixing bacteria fluid nutrient mediums, in 25 DEG C, 100r/min shaking tables shaken cultivation 2 days, must be enriched withs and consolidate
Nitrogen bacteria culture fluid, repeats enrichment culture 3 times, obtains multiple culture enrichment fixed nitrogen bacteria culture fluid, and multiple culture is enriched with into fixed nitrogen bacteria culture fluid
With the normal saline dilution that mass fraction is 0.9% to 10-5Level is diluted, fixed nitrogen bacterium solution after must diluting, fixed nitrogen bacterium solution applies after taking dilution
Nitrogen-fixing bacteria separation plating medium is distributed in, 25 DEG C of culture 2d, nitrogen-fixing bacteria bacterium colony is obtained, repeats line culture 2 times, nitrogen-fixing bacteria must be purified
Bacterium colony；
（2）Picking purifying nitrogen-fixing bacteria bacterium colony in mass ratio 1：5 are seeded in nitrogen-fixing bacteria fluid nutrient medium, 25 DEG C, 200r/min shakes
Culture 24h is swung, obtains fixed nitrogen bacteria culture fluid, by fixed nitrogen bacteria culture fluid by volume 1：5 are seeded to DF culture medium shaken cultivation 18h,
Nutrient solution, by nutrient solution by volume 1：50 are seeded in ADF culture mediums the shaken cultivation 24h under the conditions of 25 DEG C, and repetition connects
Kind culture 2 times, obtains deaminase positive bacteria culture fluid, centrifuges, takes precipitation, dries, obtains deaminase positive bacteria；
（3）Take sludge in mass ratio 1：10 mix with sterilized water, 25 DEG C, 120r/min shaken cultivation 3h, stand, and obtain sludge suspension
Liquid, 2h is stood, takes sludge suspension supernatant in mass ratio 1：In 10 inoculation desulfurization bacterium enriched mediums, 25 DEG C, 120r/min
Enrichment culture 2 days, obtains enrichment culture liquid, and enrichment culture liquid is diluted into 10-5Dilution level, desulfurization bacterium bacterium solution after must diluting, will
Desulfurization bacterium bacterium solution in mass ratio 1 after dilution：10 are seeded to desulfurization bacterium screening and culturing medium, and 25 DEG C are cultivated 2 days, obtain desulfurization bacterium bacterium colony,
The maximum desulfurization bacterium bacterium colony streak inoculation in picking bacterium footpath is cultivated 2 days for 25 DEG C to desulfurization bacterium screening and culturing medium, repeats line culture 2
It is secondary, obtain purifying desulfurization bacterium；
（4）By EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium in mass ratio 5：
2：1：4：4 is well mixed, obtains mixing decomposing microbial inoculum, by mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran in mass ratio 5：7：8：4 is mixed
Close uniformly, obtain biological decomposing agent.
Example 2
Pedotheque：It is derived from the field-crop rhizosphere in growth period.
Desulfurization bacterium screening sample：It is derived from the sludge of sewage deposited bottom.
Decomposing agent base starting material：Wood chip, rice husk, chicken manure, rice bran.
Decomposed fermented bacterium：EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte.
Desulphurization Strains screening sample：Activated sludge and anaerobic sludge selected from sewage treatment plant.
Nitrogen-fixing bacteria fluid nutrient medium：According to the mass fraction, 15 parts of sucrose, K are taken2HPO4·3H2O0.5 parts, NaCl0.5 parts,
CaCO32 parts, MgSO4·7H2O0.4 parts, 0.8 part of yeast extract, 1000 parts of distilled water, pH 7.2.
Nitrogen-fixing bacteria separate plating medium：Add 25 parts of agar into nitrogen-fixing bacteria fluid nutrient medium, other components are constant.
DF culture mediums：According to the mass fraction, KH is taken2PO46 parts, Na2HPO410 parts, MgSO4·7H2O0.5 parts, glucose 6
Part, 5 parts of sodium gluconate, 2 parts of citric acid, (NH4)2SO44 parts, 0.2 part of one solution of component, 0.2 part of two solution of component, water 1000
Part, pH 7. 4.Wherein component one is according to the mass fraction, to take H3BO30.03 part, MnSO4·H2O0.02 parts, ZnSO4·
7H2O0.3 parts, CuSO4·5H2O0.09 parts, MoO30.02 part, 100 parts of water；Component two is by FeSO47H2O and sterile purified water
In mass ratio 1：100 is well mixed, produces.
ADF culture mediums：By isolated purifying nitrogen-fixing bacteria in mass ratio 1：30 are seeded to and are free of (NH4 )2SO4Component is gone out
In bacterium DF culture mediums, ADF culture mediums are obtained, pH is 7. 5.
Desulfurization bacterium enriched medium：According to the mass fraction, 12 parts of peptone, 7 parts of beef extract, 8 parts of sodium chloride, water 1000 are taken
Part, pH 7.2.
Desulfurization bacterium screening and culturing medium：According to the mass fraction, KH is taken2PO45 parts, 18 parts of peptone, 5 parts of beef extract,
Na2HPO4·12H2O5 parts, 5 parts of sodium chloride, 25 parts of agar, 1000 parts of water, pH 7.2.
A kind of preparation method of biological decomposing agent, comprises the following steps：
（1）Soil sampling in mass ratio 1：9 are well mixed with sterilized water, shaking table vibration 20min, obtain soil sample mixed liquor, soil sample is mixed
Close liquid by volume 1：8 add in nitrogen-fixing bacteria fluid nutrient mediums, in 28 DEG C, 100r/min shaking tables shaken cultivation 3 days, must be enriched withs and consolidate
Nitrogen bacteria culture fluid, repeats enrichment culture 3 times, obtains multiple culture enrichment fixed nitrogen bacteria culture fluid, and multiple culture is enriched with into fixed nitrogen bacteria culture fluid
With the normal saline dilution that mass fraction is 0.9% to 10-5Level is diluted, fixed nitrogen bacterium solution after must diluting, fixed nitrogen bacterium solution applies after taking dilution
Nitrogen-fixing bacteria separation plating medium is distributed in, 29 DEG C of culture 3d, nitrogen-fixing bacteria bacterium colony is obtained, repeats line culture 3 times, obtain purifying fixed nitrogen
Bacterium bacterium colony；
（2）Picking purifying nitrogen-fixing bacteria bacterium colony in mass ratio 1：5 are seeded in nitrogen-fixing bacteria fluid nutrient medium, 30 DEG C, 200r/min shakes
Culture 36h is swung, obtains fixed nitrogen bacteria culture fluid, by fixed nitrogen bacteria culture fluid by volume 1：5 are seeded to DF culture medium shaken cultivation 24h,
Nutrient solution, by nutrient solution by volume 1：50 are seeded in ADF culture mediums the shaken cultivation 48h under the conditions of 30 DEG C, and repetition connects
Kind culture 3 times, obtains deaminase positive bacteria culture fluid, centrifuges, takes precipitation, dries, obtains deaminase positive bacteria；
（3）Take sludge in mass ratio 1：10 mix with sterilized water, 30 DEG C, 120r/min shaken cultivation 5h, stand, and obtain sludge suspension
Liquid, 4h is stood, takes sludge suspension supernatant in mass ratio 1：In 10 inoculation desulfurization bacterium enriched mediums, 30 DEG C, 120r/min
Enrichment culture 3 days, obtains enrichment culture liquid, and enrichment culture liquid is diluted into 10-5Dilution level, desulfurization bacterium bacterium solution after must diluting, will
Desulfurization bacterium bacterium solution in mass ratio 1 after dilution：10 are seeded to desulfurization bacterium screening and culturing medium, and 30 DEG C are cultivated 3 days, obtain desulfurization bacterium bacterium colony,
The maximum desulfurization bacterium bacterium colony streak inoculation in picking bacterium footpath is cultivated 3 days for 30 DEG C to desulfurization bacterium screening and culturing medium, repeats line culture 3
It is secondary, obtain purifying desulfurization bacterium；
（4）By EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium in mass ratio 5：
2：1：4：4 is well mixed, obtains mixing decomposing microbial inoculum, by mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran in mass ratio 5：7：8：4 is mixed
Close uniformly, obtain biological decomposing agent.
Example 3
Pedotheque：It is derived from the field-crop rhizosphere in growth period.
Desulfurization bacterium screening sample：It is derived from the sludge of sewage deposited bottom.
Decomposing agent base starting material：Wood chip, rice husk, chicken manure, rice bran.
Decomposed fermented bacterium：EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte.
Desulphurization Strains screening sample：Activated sludge and anaerobic sludge selected from sewage treatment plant.
Nitrogen-fixing bacteria fluid nutrient medium：According to the mass fraction, 12.5 parts of sucrose, K are taken2HPO4·3H2O0.3 parts, NaCl0.3
Part, CaCO31.5 parts, MgSO4·7H2O0.3 parts, 0.7 part of yeast extract, 1000 parts of distilled water, pH 7.0.
Nitrogen-fixing bacteria separate plating medium：Add 22.5 parts of agar into nitrogen-fixing bacteria fluid nutrient medium, other components are constant.
DF culture mediums：According to the mass fraction, KH is taken2PO45 parts, Na2HPO48 parts, MgSO4·7H20.3 part of O, glucose 4
Part, 4 parts of sodium gluconate, 1.5 parts of citric acid, (NH4)2SO43 parts, 0.15 part of one solution of component, 0.15 part of two solution of component, water
1000 parts, pH 7. 2.Wherein component one is according to the mass fraction, to take H3BO30.02 part, MnSO4·H2O0.01 parts, ZnSO4·
7H2O0.2 parts, CuSO4·5H2O0.08 parts, MoO30.015 part, 100 parts of water；Component two is by FeSO47H2O and sterile purified water
In mass ratio 1：100 is well mixed, produces.
ADF culture mediums：By isolated purifying nitrogen-fixing bacteria in mass ratio 1：30 are seeded to and are free of (NH4 )2SO4Component is gone out
In bacterium DF culture mediums, ADF culture mediums, pH 7.2 are obtained.
Desulfurization bacterium enriched medium：According to the mass fraction, 11 parts of peptone, 6 parts of beef extract, 6.5 parts of sodium chloride, water are taken
1000 parts, pH 7.0.
Desulfurization bacterium screening and culturing medium：According to the mass fraction, KH is taken2PO43.5 parts, 16.5 parts of peptone, 4 parts of beef extract,
Na2HPO4·12H2O3.5 parts, 4 parts of sodium chloride, 22.5 parts of agar, 1000 parts of water, pH 7.0.
A kind of preparation method of biological decomposing agent, comprises the following steps：
（1）Soil sampling in mass ratio 1：9 are well mixed with sterilized water, shaking table vibration 20min, obtain soil sample mixed liquor, soil sample is mixed
Close liquid by volume 1：8 add in nitrogen-fixing bacteria fluid nutrient mediums, in 26 DEG C, 100r/min shaking tables shaken cultivation 2.5 days, must be enriched with
Fixed nitrogen bacteria culture fluid, repeats enrichment culture 3 times, obtains multiple culture enrichment fixed nitrogen bacteria culture fluid, and multiple culture is enriched with into nitrogen-fixing bacteria culture
The normal saline dilution that liquid mass fraction is 0.9% is to 10-5Level is diluted, fixed nitrogen bacterium solution after must diluting, takes fixed nitrogen bacterium solution after dilution
Nitrogen-fixing bacteria separation plating medium is coated on, 27 DEG C of culture 2.5d, nitrogen-fixing bacteria bacterium colony is obtained, repeats line culture 2 times, must purify solid
Nitrogen bacterium bacterium colony；
（2）Picking purifying nitrogen-fixing bacteria bacterium colony in mass ratio 1：5 are seeded in nitrogen-fixing bacteria fluid nutrient medium, 27 DEG C, 200r/min shakes
Culture 30h is swung, obtains fixed nitrogen bacteria culture fluid, by fixed nitrogen bacteria culture fluid by volume 1：5 are seeded to DF culture medium shaken cultivation 21h,
Nutrient solution, by nutrient solution by volume 1：50 are seeded in ADF culture mediums the shaken cultivation 36h under the conditions of 27 DEG C, and repetition connects
Kind culture 2 times, obtains deaminase positive bacteria culture fluid, centrifuges, takes precipitation, dries, obtains deaminase positive bacteria；
（3）Take sludge in mass ratio 1：10 mix with sterilized water, 27 DEG C, 120r/min shaken cultivation 4h, stand, and obtain sludge suspension
Liquid, 3h is stood, takes sludge suspension supernatant in mass ratio 1：In 10 inoculation desulfurization bacterium enriched mediums, 27 DEG C, 120r/min
Enrichment culture 2.5 days, obtains enrichment culture liquid, and enrichment culture liquid is diluted into 10-5Dilution level, desulfurization bacterium bacterium solution after must diluting,
Desulfurization bacterium bacterium solution in mass ratio 1 after diluting：10 are seeded to desulfurization bacterium screening and culturing medium, and 27 DEG C are cultivated 2.5 days, obtain desulfurization bacterium bacterium
Fall, the maximum desulfurization bacterium bacterium colony streak inoculation in picking bacterium footpath is cultivated 2.5 days for 27 DEG C to desulfurization bacterium screening and culturing medium, repeats line training
Support 2 times, obtain purifying desulfurization bacterium；
（4）By EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium in mass ratio 5：
2：1：4：4 is well mixed, obtains mixing decomposing microbial inoculum, by mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran in mass ratio 5：7：8：4 is mixed
Close uniformly, obtain biological decomposing agent.
Reference examples：The biological decomposing agent of company of Beijing production.
Using pig manure as fermenting raw materials organic fertilizer, sent out respectively using the decomposing agent of the present invention and the decomposing agent of reference examples
Ferment, wherein using decomposing agent 8kg per 1000kg raw materials.Fermentation temperature is controlled at 50 ~ 65 DEG C, after the completion of fermentation, calculates each processing
The content of Nitrogen.In addition, the 1st, 3,5,7 day in fermentation respectively, gathers each processing ambient gas respectively, detection is therein
The reduction content of ammonia and hydrogen sulfide, it the results are shown in Table 1, table 2, table 3.
Table 1：The reduction content of ammonia, hydrogen sulfide and foul smell discharge.
| Test event | Example 1 | Example 2 | Example 3 | Reference examples |
| Ammonia reduces % | 84 | 82 | 82 | 38~42 |
| Hydrogen sulfide reduces % | 81 | 84 | 81 | 37~43 |
| Foul smell reduces % | 74 | 76 | 77 | 32~36 |
Table 2：Each component content in fertilizer.
| Test event | Example 1 | Example 2 | Example 3 | Reference examples |
| Nitrogen content % | 51 | 54 | 58 | 18~26 |
| Total nutrient % | 9.4 | 9.2 | 9.6 | 3.4~4.2 |
| Organic matter % | 21 | 16 | 14 | 6.2~7.2 |
Table 3：Decomposed effect（d）.
| Test event | Example 1 | Example 2 | Example 3 | Reference examples |
| The time required to softening | 6 | 6 | 5 | 11~15 |
| Decomposed required time | 21 | 22 | 21 | 40~48 |
Summary, the decomposed effect of biological decomposing agent of the invention is good and digest process in can reduce the row of foul smell well
Put, be worth of widely use.
Claims (8)
1. a kind of preparation method of biological decomposing agent, it is characterised in that the preparation method comprises the following steps：
Soil sampling is well mixed with sterilized water, shaking table vibration, obtains soil sample mixed liquor, and soil sample mixed liquor is added into the training of fixed nitrogen bacteria liquid
Support and cultivated in base, fixed nitrogen bacteria culture fluid must be enriched with, repeat enrichment culture 3 times, obtain multiple culture enrichment fixed nitrogen bacteria culture fluid, by multiple training
Normal saline dilution that enrichment fixed nitrogen bacteria culture fluid mass fraction is 0.9% is supported to 10-5Dilution level, fixed nitrogen bacterium solution after must diluting,
Fixed nitrogen bacterium solution is coated on nitrogen-fixing bacteria separation plating medium after taking dilution, culture, obtains nitrogen-fixing bacteria bacterium colony, repeats line culture 2 ~ 3
It is secondary, nitrogen-fixing bacteria bacterium colony must be purified；
Picking purifying nitrogen-fixing bacteria colony inoculation is cultivated into nitrogen-fixing bacteria fluid nutrient medium, obtains fixed nitrogen bacteria culture fluid, nitrogen-fixing bacteria are trained
Nutrient solution is seeded to DF culture medium shaken cultivations, obtains nutrient solution, and nutrient solution is seeded in ADF culture mediums and cultivated, repeated inoculation training
Support 2 ~ 3 times, obtain deaminase positive bacteria culture fluid, centrifuge, take precipitation, dry, obtain deaminase positive bacteria；
Sludge is taken to be mixed with sterilized water, culture, sludge suspension is obtained, is stood, takes the supernatant of sludge suspension, is inoculated with desulfurization
Enrichment culture in bacterium enriched medium, enrichment culture liquid is obtained, enrichment culture liquid is diluted to 10-5Dilution level, after must diluting take off
Sulphur bacterium bacterium solution, desulfurization bacterium bacterium solution after dilution is seeded to desulfurization bacterium screening and culturing medium culture, obtains desulfurization bacterium bacterium colony, picking bacterium footpath is most
Big desulfurization bacterium bacterium colony streak inoculation repeats line culture 2 ~ 3 times, obtains purifying desulfurization bacterium to desulfurization bacterium screening and culturing medium culture；
EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, acc deaminase positive bacteria, purifying desulfurization bacterium are well mixed, obtained mixed
Decomposing microbial inoculum is closed, mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran are well mixed, obtain biological decomposing agent.
2. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（1）Middle soil-like
Product are derived from the potato rhizosphere soil in growth period；The formula of nitrogen-fixing bacteria fluid nutrient medium for according to the mass fraction, take sucrose 10 ~
15 parts, K2HPO4·3H20.2 ~ 0.5 part of O, NaCl0.2 ~ 0.5 part, CaCO31 ~ 2 part, MgSO4·7H20.2 ~ 0.4 part of O, yeast
0.5 ~ 0.8 part of cream, 1000 parts of distilled water, pH 7. 0 ± 0.2；Nitrogen-fixing bacteria separate plating medium：To nitrogen-fixing bacteria Liquid Culture
Add 20 ~ 25 parts of agar in base, other components are constant.
3. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（1）Middle soil sample with
The mass ratio of sterilized water is 1：9；The volume ratio of soil sample mixed liquor and nitrogen-fixing bacteria fluid nutrient medium is 1：8, enrichment nitrogen-fixing bacteria culture
The condition of culture of liquid is 25 ~ 28 DEG C, 100r/min shaking tables shaken cultivation 2 ~ 3 days；Purify the condition of culture of nitrogen-fixing bacteria bacterium colony for 25 ~
29 DEG C of 2 ~ 3d of culture.
4. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（2）Middle DF cultures
The formula of base is according to the mass fraction, to take KH2PO44 ~ 6 parts, Na2HPO46 ~ 10 parts, MgSO4·7H20.2 ~ 0.5 part of O, glucose
2 ~ 6 parts, 3 ~ 5 parts of sodium gluconate, 1 ~ 2 part of citric acid, (NH4) 2SO42 ~ 4 parts, 0.1 ~ 0.2 part of one solution of component, component two
0.1 ~ 0.2 part of solution, 1000 parts of water, pH 7. 2 ± 0.2, wherein component one are according to the mass fraction, to take H3BO30.01~0.03
Part, MnSO4·H20.01 ~ 0.02 part of O, ZnSO4·7H20.1 ~ 0.3 part of O, CuSO4·5H20.07 ~ 0.09 part of O, MoO3
0.01 ~ 0.02 part, 100 parts of water；Component two is by FeSO47H2O and sterile purified water in mass ratio 1：100 is well mixed, i.e.,
；The formula of ADF culture mediums is by isolated purifying nitrogen-fixing bacteria in mass ratio 1：30 are seeded to and are free of (NH4 ) 2SO4Component
Sterilizing DF culture mediums in, obtain ADF culture mediums, pH is 7 ~ 7. 5.
5. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（2）Middle purifying is solid
The mass ratio of nitrogen bacterium bacterium colony and nitrogen-fixing bacteria fluid nutrient medium is 1：5, condition of culture be 25 ~ 30 DEG C, 200r/min shaken cultivations 24 ~
36h；The volume ratio of fixed nitrogen bacteria culture fluid and DF culture mediums is 1：5；Nutrient solution by volume 1：50 are seeded in ADF culture mediums
24 ~ 48h of shaken cultivation under the conditions of 25 ~ 30 DEG C.
6. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（3）Middle desulfurization bacterium
Screening sample is derived from the sludge of sewage deposited bottom；The formula of desulfurization bacterium enriched medium is according to the mass fraction, to take peptone
10 ~ 12 parts, 5 ~ 7 parts of beef extract, 5 ~ 8 parts of sodium chloride, 1000 parts of water, pH is 7.0 ± 0.2；The formula of desulfurization bacterium screening and culturing medium
According to the mass fraction, to take KH2PO42 ~ 5 parts, 15 ~ 18 parts of peptone, 3 ~ 5 parts of beef extract, Na2HPO4·12H202 ~ 5 part, chlorination
3 ~ 5 parts of sodium, 20 ~ 25 parts of agar, 1000 parts of water, pH are 7.0 ± 0.2.
7. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（3）Middle sludge with
The mass ratio of sterilized water is 1：10, condition of culture is 25 ~ 30 DEG C, 3 ~ 5h of 120r/min shaken cultivations；Sludge suspension supernatant
Mass ratio with desulfurization bacterium enriched medium is 1：10, condition of culture is 25 ~ 30 DEG C, 120r/min enrichment cultures 2 ~ 3 days.
8. the preparation method of biological decomposing agent according to claim 1, it is characterised in that the step（4）Middle mixing is rotten
Ripe microbial inoculum is EM bacterium, false Ruan's silk saccharomycete, Gymboree zymophyte, deaminase positive bacteria, purifying desulfurization bacterium in mass ratio 5：2：1：
4：4 is well mixed, and biological decomposing agent is mixing decomposing microbial inoculum, chicken manure, rice husk, rice bran in mass ratio 5：7：8：4 is well mixed.
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