CN104744091A - Method for preparing organic material decomposing microbial inoculum based on white trichoderma and bacillus subtilis - Google Patents
Method for preparing organic material decomposing microbial inoculum based on white trichoderma and bacillus subtilis Download PDFInfo
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- CN104744091A CN104744091A CN201510136232.4A CN201510136232A CN104744091A CN 104744091 A CN104744091 A CN 104744091A CN 201510136232 A CN201510136232 A CN 201510136232A CN 104744091 A CN104744091 A CN 104744091A
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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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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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
- 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
Abstract
The invention discloses a method for preparing an organic material decomposing microbial inoculum based on white trichoderma and bacillus subtilis. The method comprises the following steps: performing cultivation and proliferation on the white trichoderma and the bacillus subtilis through a multistage culture method; after the maximum activity concentration is reached, adding the microorganism in the processed animal waste according to a special inoculation size and fermenting to obtain the organic material decomposing microbial inoculum. By adopting the organic material decomposing microbial inoculum manufactured by beneficial bacteria and fungus, a macromolecule protein and the carbohydrate in the waste and the straw can be decomposed into micromolecule substances, the elements such as phosphorus and potassium are dissociated and the micro metal element is separated out so that the organic material decomposing microbial inoculum can be absorbed and utilized by a plant and the growth speed of the plant can be quickened; simultaneously, the harm on the environment and the humankind due to hard decomposition of the macromolecule organic matters in the waste such as the protein and the cellulose can be avoided.
Description
Technical field
The present invention relates to a kind of organic material composting microbial inoculum, specifically, relate to a kind of preparation method of the organic material composting microbial inoculum based on white Trichoderma [numbering: CGMCC3.2941] and subtilis [numbering: CGMCC1.4255].
Background technology
Flourish along with modern agriculture in recent years, the agriculture organic materials wastes such as the Amount of poultry excrements of aquaculture and the stalk of plant husbandry also increase year by year.If these organic materials wastes, without the harmless treatment of microorganism, can produce a large amount of waste gas, waste water and waste residue, cause huge environmental pollution, and have a strong impact on the life health of people.The traditional method of process organic materials waste is as fertilizer after ight soil natural packing being fermented or diluting with soil, water, but these methods fundamentally can not solve the pollution problem of ight soil to environment, and nitrogen, phosphorus, carbon, potassium and the trace element in organic materials waste is not absorbed fully by farm crop and utilize, the ight soil in organic materials waste contains infecting both domestic animals and human cause of disease bacterium and is not also entirely killed.
Summary of the invention
The object of the invention is to the defect overcoming the existence of above-mentioned technology, thering is provided a kind of preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis, is a kind ofly carry out the organic material composting microbial inoculum that agriculture organic materials waste harmless treatment makes microbial fertilizer.
Its concrete technical scheme is:
Based on a preparation method for the organic material composting microbial inoculum of white Trichoderma and subtilis, comprise the steps:
Step one: cultivate white Trichoderma:
(1) in solid medium, slant culture is carried out to white Trichoderma, at 28-30 DEG C, cultivate 72h, obtain first order seed bacterium;
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 28-30 DEG C of temperature, cultivate 3-5 days, obtain secondary seed bacterium;
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 3 days under 28-30 DEG C of condition, vibration velocity is 5-8 time/min; Again the bacterium liquid in liquid triangular flask is inoculated in the fermentor tank of same liquid substratum by 1-2%, 28-30 DEG C of bottom fermentation 3-5 days, obtains three grades of kind daughter bacterias;
(4) tank is by fermentation cultivated the white Trichoderma three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% ~ 45% of fermention medium, first pH6.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated into aerobic fermentation in the ight soil for first processing by 1-2%, temperature stirs cooling more than 45 degree, fermentation 6-8 days, every day, control temperature was at 30-33 DEG C, obtained stand-by white Trichoderma bacterium liquid fermentation ight soil;
Step 2: cultivate subtilis:
(1) in solid medium, slant culture is carried out to subtilis, temperature 37-38 DEG C, under the condition of humidity 60%, ventilation lucifuge cultivates 36-48h, obtains first order seed bacterium;
(2) first order seed bacterium is gone down to posterity to on the solid slant culture base of filling a prescription identical in (1) through inclined-plane, at 37 DEG C of temperature, cultivate 24h, obtain secondary seed bacterium;
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 36-48h under 37 DEG C of conditions, vibration velocity is 5-8 time/min; Again the bacterium liquid in liquid triangular flask is inoculated in the fermentor tank of same liquid substratum by 1-2%, aerated culture 3-5 days at 35-38 DEG C, obtains three grades of kind daughter bacterias;
(4) tank is by fermentation cultivated the subtilis three grades of kind daughter bacterias obtained to be inoculated into and to consist of in the animal excrement after the humidity process of 30% ~ 45% containing substratum, first pH7.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, stirring cooling is carried out after ventilation 24h, ferment about 5 days time, every day, control temperature was at 30-40 DEG C, obtained stand-by subtilis bacterium liquid;
Step 3: the stand-by subtilis bacterium liquid that stand-by white Trichoderma bacterium liquid step one obtained and step 2 obtain, add in animal excrement by the add-on of 20-30% and 10-15% respectively, and add the sodium carbonate of animal excrement amount 0.1-0.22% simultaneously, obtain microbial fertilizer through there being the alternately fermentation of aerobic and anaerobism.
Preferably, the formula of step one (1) and the solid medium described in (2) is: glucose 2%, sodium-chlor 0.1%, peptone 1-2%, yeast extract paste 0.5%, agar powder 2%, surplus is water, sterilizing 15min under pH 6.5,115 DEG C of conditions.
Preferably, the formula of the liquid nutrient medium described in step one (3) is:
Glucose 2%, sodium-chlor 0.1%, peptone 1-2%, yeast extract paste 0.5%, surplus is water, pH6.5, sterilizing 20min under 115 DEG C of conditions.
Preferably, the bacterium number of stand-by white Trichoderma bacterium liquid that step one (4) obtains is 400-600 hundred million/mL.
Preferably, the formula of solid medium described in step 2 (1) and (2) is:
Peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, agar powder 2%, pH7.5, sterilizing 20min under 121 DEG C of conditions.
Preferably, the formula of the liquid fermentation medium described in step 2 (3) is:
Peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, surplus is water, pH7.5,121 DEG C, sterilizing 20min.
Preferably, the bacterium number of stand-by subtilis bacterium liquid that step 2 (4) obtains is 2.4-4.8 × 10^8/mL.
Preferably, described organic material composting microbial inoculum with stalk be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1.5%.
Preferably, described organic material composting microbial inoculum with feces of livestock and poultry be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1%.
Preferably, described organic material composting microbial inoculum with stalk and feces of livestock and poultry for raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 1% and 1.5%.
Compared with prior art, beneficial effect of the present invention is:
Organic material composting microbial inoculum of the present invention utilizes beneficial bacteria and fungi to carry out special processing to ight soil and stalk, macro-molecular protein in ight soil and stalk and carbohydrate breakdown are become small-molecule substance, inorganic molecules material even, and the element such as phosphorus, potassium is dissociated, micro-metals is separated out, thus is easily absorbed by plants and utilizes, and plant growth rate is accelerated, disease resistance strengthens, and product quality improves, mouthfeel promotes; Simultaneously, decrease larger molecular organics matter in ight soil and stalk as the harm to environment and the mankind owing to not easily decomposing such as protein, Mierocrystalline cellulose, decrease the discharge of the three wastes (waste water,waste gas and industrial residue), also eradicate the potential pathogenic harm of the infecting both domestic animals and human cause of disease bacterium in ight soil.
Embodiment
Below in conjunction with specific embodiment, technical scheme of the present invention is described in more detail.
The preparation method of a kind of organic material composting microbial inoculum based on white Trichoderma and subtilis involved in the present invention, under certain condition, the amylase that the proteolytic enzyme utilizing white Trichoderma to produce and subtilis produce and cellulase, pre-treatment in early stage is carried out to the ight soil of animal and stalk, ight soil acts on mutually with stalk and specific function yeast, carry out the alternately fermentation of aerobic and anaerobism, to reduce the pollution to environment such as organic in ight soil and harmful levels of pathogens.Meanwhile, after fermentation, the ight soil of animal is easily fully absorbed by farm crop and utilizes, thus promotes the growth of plant.
White Trichoderma bacterium liquid and subtilis bacterium liquid all entered multistage cultivation, plant daughter bacteria to humans and animals without any harm, biological nature typical case, benefit materials (proteolytic enzyme, cellulase, amylase etc.) can be produced, secretory volume is large, energetic, long service life, be convenient at Laboratories Accession and the bacterial classification used that goes down to posterity, secondary seed bacterium is bred, reaches certain quantity and obtain by first order seed bacterium Secondary Culture, and secondary seed bacterium is carried out cultivations and rises in value and reach production and application requirement and obtain by three grades of kind daughter bacterias.Three grades of kind daughter bacterias, after animal excrement fermentative processing, obtain stand-by bacterium liquid, finally make organic material composting microbial inoculum.This organic material composting microbial inoculum adds the alternately fermentation through aerobic and anaerobism in animal excrement, obtains organic material composting microbial inoculum.This organic material composting microbial inoculum process animal excrement, make that it becomes loose, free from extraneous odour, to the stingless excitation of people on sense organ, organic macromolecule in ight soil changes organic little macromolecular substance into, the element such as phosphorus, potassium dissociates, micro-metals is therefrom separated out, and makes the ight soil be harmful to reach the object of microorganism harmless treatment.
Embodiment 1
Described organic material composting microbial inoculum concrete steps are as follows:
Step one: cultivate white Trichoderma:
(1) in solid medium, slant culture is carried out to white Trichoderma, cultivate 3 days at 28-30 DEG C, obtain first order seed bacterium;
The formula of described solid medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, sterilizing 15min under agar powder 2%, pH 6.5,115 DEG C of condition.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 28-30 DEG C of temperature, cultivate 3-5 days, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 3 days under 28-30 DEG C of condition, vibration velocity is 5-8 time/min; Again the bacterium liquid in liquid triangular flask is inoculated in the fermentor tank of same liquid substratum by 1-2%, 28-30 DEG C of bottom fermentation 3-5 days, obtains three grades of kind daughter bacterias;
The formula of described liquid nutrient medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, pH6.5, sterilizing 20min under 115 DEG C of conditions.
(4) tank is by fermentation cultivated the white Trichoderma three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% ~ 45% of fermention medium, first pH6.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1-2%, stirring cooling is carried out after ventilation 24h, fermentation 6-8 days, every day, control temperature was at 30-33 DEG C, obtain stand-by white Trichoderma bacterium liquid, bacterium total count is 400-600 hundred million/mL.
Step 2: cultivate subtilis:
(1) in solid medium, slant culture is carried out to subtilis, temperature 36-38 DEG C, under the condition of humidity 60%, ventilation lucifuge cultivates 48h, obtains first order seed bacterium;
The formula of described solid medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, agar powder 2%, pH7.5, sterilizing 20min under 121 DEG C of conditions.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 37 DEG C of temperature, cultivate 24h, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 48h under 37 DEG C of conditions, vibration velocity is 8 times/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1-2% again, shaking culture of ventilating at 35 DEG C 5 days, vibration velocity is 8 times/min, obtains three grades of kind daughter bacterias;
The formula of described liquid fermentation medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, and surplus is water, pH7.5,121 DEG C, sterilizing 20min.
(4) tank is by fermentation cultivated the subtilis three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 45% of fermention medium, first pH7.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 2%, stirring cooling is carried out after ventilation 24h, ferment 5 days, every day, control temperature was at 40 DEG C, obtain stand-by subtilis bacterium liquid, bacterium total count bacterium number is 2.4-4.8 × 1,0^8 hundred million/mL.
Step 3: the stand-by subtilis bacterium liquid that stand-by white Trichoderma bacterium liquid step one obtained and step 2 obtain, add in animal excrement by the add-on of 30% and 15% respectively, and adding the sodium carbonate of animal excrement amount 2% simultaneously, the alternately fermentation through aerobic and anaerobism obtains organic material composting microbial inoculum.
Embodiment 2
Described organic material composting microbial inoculum concrete steps are as follows:
Step one: cultivate white Trichoderma:
(1) in solid medium, slant culture is carried out to white Trichoderma, cultivate 3 days at 28 DEG C, obtain first order seed bacterium;
The formula of described solid medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, sterilizing 15min under agar powder 2%, pH 6.5,115 DEG C of condition.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 28 DEG C of temperature cultivation 3 days, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 3 days under 28 DEG C of conditions, vibration velocity is 5 times/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1% again, 28 DEG C of bottom fermentations 3 days, obtain three grades of kind daughter bacterias;
The formula of described liquid nutrient medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, and surplus is water, pH6.5, sterilizing 20min under 115 DEG C of conditions.
(4) tank is by fermentation cultivated the white Trichoderma three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% of fermention medium, first pH6.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1-2%, stirring cooling is carried out after ventilation 24h, ferment 6 days, every day, control temperature was at 45 DEG C, obtain stand-by white Trichoderma bacterium liquid, bacterium total count bacterium number is 400-600 hundred million/mL.
Step 2: cultivate subtilis:
(1) in solid medium, carry out slant culture to subtilis, under the condition of temperature 37 DEG C, humidity 60%, ventilation lucifuge cultivates 48h, obtains first order seed bacterium;
The formula of described solid medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, agar powder 2%, pH7.5, sterilizing 20min under 121 DEG C of conditions.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 37 DEG C of temperature, cultivate 24h, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 48h under 37 DEG C of conditions, vibration velocity is 5 times/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1-2% again, shaking culture of ventilating at 35 DEG C 3 days, vibration velocity is 5 times/min, obtains three grades of kind daughter bacterias;
The formula of described liquid fermentation medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, and surplus is water, pH7.5,121 DEG C, sterilizing 15min.
(4) tank is by fermentation cultivated the subtilis three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% of fermention medium, first pH7.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1%, stirring cooling is carried out after ventilation 24h, ferment 5 days, every day, control temperature was at 30-40 DEG C, obtain stand-by subtilis bacterium liquid, bacterium number is 2.4-4.8 × 10^8/mL.
Step 3: the stand-by subtilis bacterium liquid that stand-by white Trichoderma bacterium liquid step one obtained and step 2 obtain, add in animal excrement by the add-on of 20% and 10% respectively, and adding the sodium carbonate of animal excrement amount 1% simultaneously, the alternately fermentation through aerobic and anaerobism obtains organic material composting microbial inoculum.
Embodiment 3
Described organic material composting microbial inoculum concrete steps are as follows:
Step one: cultivate white Trichoderma:
(1) in solid medium, slant culture is carried out to white Trichoderma, cultivate 3 days at 28 DEG C, obtain first order seed bacterium;
The formula of described solid medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, agar powder 2%, and surplus is water, sterilizing 15min under pH 6.5,115 DEG C of conditions.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 29 DEG C of temperature cultivation 4 days, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 3 days under 29 DEG C of conditions, vibration velocity is 6 times/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1.5% again, 29 DEG C of bottom fermentations 4 days, obtain three grades of kind daughter bacterias;
The formula of described liquid nutrient medium is: glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, pH6.5, sterilizing 15-20min under 115 DEG C of conditions.
(4) tank is by fermentation cultivated the white Trichoderma three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 40% of fermention medium, first pH6.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1-2%, stirring cooling is carried out after ventilation 24h, ferment 7 days, every day, control temperature was at 32 DEG C, obtain stand-by white Trichoderma bacterium liquid, bacterium number is 400-600 hundred million/mL.
Step 2: cultivate subtilis:
(1) in solid medium, carry out slant culture to subtilis, under the condition of temperature 38 DEG C, humidity 60%, ventilation lucifuge cultivates 48h, obtains first order seed bacterium;
The formula of described solid medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, agar powder 2%, pH7.5, sterilizing 20min under 121 DEG C of conditions.
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 37 DEG C of temperature, cultivate 24h, obtain secondary seed bacterium.
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 48h under 37 DEG C of conditions, vibration velocity is 7 times/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1-2% again, shaking culture of ventilating at 35 DEG C 4 days, vibration velocity is 65-8 time/min, obtains three grades of kind daughter bacterias;
The formula of described liquid fermentation medium is: peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, and surplus is water, pH7.5,121 DEG C, sterilizing 20min.
(4) tank is by fermentation cultivated the subtilis three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 40% of fermention medium, first pH7.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1.5%, stirring cooling is carried out after ventilation 24h, ferment 5 days, every day, control temperature was at 35 DEG C, obtain stand-by subtilis bacterium liquid, bacterium total count bacterium number is 2.4-4.8 × 1,0^8 hundred million/mL.
Step 3: the stand-by subtilis bacterium liquid that stand-by white Trichoderma bacterium liquid step one obtained and step 2 obtain, add in animal excrement by the add-on of 25% and 12% respectively, and adding the sodium carbonate of animal excrement amount 1.5% simultaneously, the alternately fermentation through aerobic and anaerobism obtains organic material composting microbial inoculum.
Described organic material composting microbial inoculum with stalk be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1.5%; Described organic material composting microbial inoculum with feces of livestock and poultry be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1%.Described organic material composting microbial inoculum with stalk and feces of livestock and poultry for raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 1% and 1.5%;
The above; be only the present invention's preferably embodiment; protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses, the simple change of the technical scheme that can obtain apparently or equivalence are replaced and are all fallen within the scope of protection of the present invention.
Claims (10)
1., based on a preparation method for the organic material composting microbial inoculum of white Trichoderma and subtilis, it is characterized in that, comprise the steps:
Step one: cultivate white Trichoderma:
(1) in solid medium, slant culture is carried out to white Trichoderma, cultivate 3 days at 28-30 DEG C, obtain first order seed bacterium;
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 28-30 DEG C of temperature, cultivate 3-5 days, obtain secondary seed bacterium;
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 3 days under 28-30 DEG C of condition, vibration velocity is 5-8 time/min; Again the bacterium liquid in liquid triangular flask is inoculated in the fermentor tank of same liquid substratum by 1-2%, 28-30 DEG C of bottom fermentation 3-5 days, obtains three grades of kind daughter bacterias;
(4) tank is by fermentation cultivated the white Trichoderma three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% ~ 45% of fermention medium, first pH6.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1-2%, stirring cooling is carried out after ventilation 24h, fermentation 6-8 days, every day, control temperature was at 30-33 DEG C, obtained stand-by white Trichoderma bacterium liquid;
Step 2: cultivate subtilis:
(1) in solid medium, slant culture is carried out to subtilis, temperature 37-40 DEG C, under the condition of humidity 60%, ventilation lucifuge cultivates 48h, obtains first order seed bacterium;
(2) first order seed bacterium is gone down to posterity to through inclined-plane with (1) in fill a prescription on identical solid medium, at 37 DEG C of temperature, cultivate 24h, obtain secondary seed bacterium;
(3) be inoculated in liquid nutrient medium by secondary seed bacterium, be divided in liquid triangular flask, oscillation and fermentation cultivation 48h under 37 DEG C of conditions, vibration velocity is 5-8 time/min; Be inoculated in the fermentor tank of same liquid substratum by the bacterium liquid in liquid triangular flask by 1-2% again, ventilate at 35 DEG C shaking culture 3-5 days, and vibration velocity is 5-8 time/min, obtains three grades of kind daughter bacterias;
(4) tank is by fermentation cultivated the subtilis three grades of kind daughter bacterias obtained and carry out fermentative processing, the animal excrement consisted of after the humidity process of 30% ~ 45% of fermention medium, first pH7.5 is debugged to acidity sodium carbonate original in ight soil during fermentation, then bacterium liquid fermentor cultivation obtained is inoculated in the ight soil for first processing by 1-2%, stirring cooling is carried out after ventilation 24h, ferment 5 days, every day, control temperature was at 30-40 DEG C, obtained stand-by subtilis bacterium liquid;
Step 3: the stand-by subtilis bacterium liquid that stand-by white Trichoderma bacterium liquid step one obtained and step 2 obtain, add in animal excrement by the add-on of 20-30% and 10-15% respectively, and adding the sodium carbonate of animal excrement amount 1-2% simultaneously, the alternately fermentation through aerobic and anaerobism obtains the attached microbial inoculum of organic materials.
2. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, the formula of step one (1) and the solid medium described in (2) is:
Glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, sterilizing 15 minutes under agar powder 2%, pH 6.5,115 DEG C of condition.
3. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, the formula of the liquid nutrient medium described in step one (3) is:
Glucose 2%, sodium-chlor 0.1%, peptone 2%, yeast extract paste 0.5%, surplus is water, pH6.5, sterilizing 15-20min under 115 DEG C of conditions.
4. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, the bacterium number of the stand-by white Trichoderma bacterium liquid that step one (4) obtains is 400-600 hundred million/mL.
5. the microorganism fertilizer preparation method for material based on white Trichoderma and subtilis according to claim 1, is characterized in that, described in step 2 (1) and (2), the formula of solid medium is:
Peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, agar powder 2%, pH7.5, sterilizing 15-20min under 121 DEG C of conditions.
6. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, the formula of the liquid fermentation medium described in step 2 (3) is:
Peptone 2%, extractum carnis 0.5%, sodium-chlor 0.5%, dipotassium hydrogen phosphate 0.1%, surplus is water, pH7.5,121 DEG C, sterilizing 20min.
7. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, the bacterium number of the stand-by subtilis bacterium liquid that step 2 (4) obtains is 2.4-4.8 × 10^8/mL.
8. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, described organic material composting microbial inoculum with stalk be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1.5%.
9. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, described organic material composting microbial inoculum with feces of livestock and poultry be raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 2% and 1%.
10. the preparation method of the organic material composting microbial inoculum based on white Trichoderma and subtilis according to claim 1, it is characterized in that, described organic material composting microbial inoculum with stalk and feces of livestock and poultry for raw material carry out fermentative processing make organic fertilizer time, the inoculum size of cultivating three grades of kind daughter bacterias in white Trichoderma bacterium liquid and subtilis bacterium liquid process is respectively 1% and 1.5%.
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