CN102942391A - Method for producing actinomyces fertilizer by utilizing fermentation of biogas residues and biogas slurry solids - Google Patents
Method for producing actinomyces fertilizer by utilizing fermentation of biogas residues and biogas slurry solids Download PDFInfo
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- CN102942391A CN102942391A CN2012104679296A CN201210467929A CN102942391A CN 102942391 A CN102942391 A CN 102942391A CN 2012104679296 A CN2012104679296 A CN 2012104679296A CN 201210467929 A CN201210467929 A CN 201210467929A CN 102942391 A CN102942391 A CN 102942391A
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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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
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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 relates to a method for producing actinomyces fertilizer, in particular to a method for producing the actinomyces fertilizer by utilizing fermentation of biogas residues and biogas slurry solids, which aims to solve the problem that the existing method for culturing actinomyces for fertilizer is high in raw material cost. The method comprises the following steps of: 1, seed culture by tube culture: a, preparing a tube culture medium, and b, inoculating the actinomyces into the tube culture medium for culture; 2, solid spore seed culture: a, preparing a seed culture medium, b, inoculating the cultured actinomyces into the seed culture medium for culture; and 3, fermentation culture: a, preparing a fermentation raw material, and b, inoculating the disinfected fermentation raw material into the spores of the actinomyces, and performing fermentation to obtain the actinomyces fertilizer. According to the invention, methane fermenting waste-biogas residues and biogas slurry is used to replace artificially synthesized culture medium to be as a fermentation substrate, so that the cost for culturing the actinomyces can be reduced. The method disclosed by the invention is applicable to the field of microorganism.
Description
Technical field
The present invention relates to a kind of method of producing actinomycetes fertilizer.
Background technology
Actinomycetes are resource microorganisms that a class is very close with human relation and have huge applications value, not only in material cycle, play an important role, and produce ability and be celebrated with abundant secondary metabolite, but the multiple important biologically active substances such as actinomycetes metabolism generation antibiosis rope, amino acid, zymin, VITAMIN and weedicide.The nineteen twenty-nine discovery of penicillin indicates the arrival in microbiotic epoch, and the Streptomycin sulphate found of Waksman has solved at that time this social concern of tuberculosis and caused extensive concern subsequently, and has excited human interest to new anti-screening.The microbe-derived Novel physiological active substance that ends discovery in 2000 has reached 16000 kinds, wherein has more than 70% to be produced by actinomycetes.The microbiotic in actinomycetes source plays an important role in clinical disease treatment, and the antibiosis that uses clinically have 2/3 and derives from actinomycetes; The microbiotic that actinomycetes produce also is the main source of agricultural antibiotic, and the Avrmectin that produces such as Avid kyowamycin is that important desinsection and extremely snail agent, jingganmycin can be used for preventing and treating rice sheath blight disease etc.Frankia in the actinomycetes can with the non-leguminous plant symbiosis, and the same frankia can infect the host plant of different genera, for huge contribution has been made in the nitrogen supply of natural Nitrogen Cycling and agroforestry, and good application prospect arranged.
Produce bacterium and activity distribution situation from common approximately 4300 kinds of new microbe meta-bolitess of delivering at JA between 50 years of nineteen forty-seven to 1997 year.Therefore, actinomycetes are as the important Microbial resources of a class, and research has been subject to countries in the world scholars' very big attention.Doing a lot of work aspect the actinomycetes research at present, wherein the most fruitful research, the classification of actinomycetes that surely belongs to the biologically active substance in actinomycetes source learned research and actinomycetes genetics research etc.Classification of actinomycetes learn research not only can for put in the saline-alkali environment actinomycetes polyphase sort and exempt from the development and use of culture studies line bacterium resource and effectively the harmful actinomycetes of control theoretical direction is provided, and can provide material and foundation for studying actinomycetic phylogenetic relationship, enrich the content of biodiversity research.Thereby classification of actinomycetes research has theoretical and actual double meaning.
Generally use the artificial synthetic medium for actinomycetic cultivation at present, be rich in a large number of nutrients in the substratum, but raw materials cost is relatively high, be applied to agricultural fertilizer with having improved production cost consumption in the actinomycetes production process.
Summary of the invention
The present invention will solve existing fertilizer with the high problem of actinomycetes cultural method raw materials cost, and a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer is provided
A kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer of the present invention, carry out according to the following steps: one, test tube is cultivated seed culture: a, preparation test-tube culture medium: take by weighing 2～2.5 parts of starch, 0.1～0.2 part in saltpetre, 0.03～0.05 part of potassium primary phosphate, 0.001～0.002 part in sal epsom, 2～2.2 parts in agar and 95～96 parts of water by mass fraction and mix, then carry out high-temperature sterilization, namely obtain test-tube culture medium; B, actinomycetes being inoculated in the test-tube culture medium, is that the inclined-plane leaves standstill cultivates 48～96h under 28～32 ℃ the condition in culture temperature, then use 1mL sterilized water wash-out test tube slant culture after, obtain elutriant; Two, solid spore seed culture: c, preparation seed culture medium: be 1 with millet in mass ratio with water: the ratio of (1～2) is mixed, and then immersion 20～24h carries out high-temperature sterilization, namely obtains seed culture medium; D, the elutriant in the step 1 being inoculated in the seed culture medium, is that shake-flask culture 72～120h obtains actinospore bacterium liquid under 28～32 ℃ the condition in culture temperature; The preparation of three, fermentation culture: e, fermentation raw material: take by weighing 70～90 parts of natural pond slags, 10～30 parts of wheat bran skins and CaCO by mass fraction
30.5～2 parts mix, and namely obtain fermentation raw material, the water content of regulating fermentation raw material with natural pond liquid is 40～45%, then carries out high-temperature sterilization; F, be in 0.5～5% fermentation raw material that is inoculated in after the sterilization with the actinospore bacterium liquid in the step 2 by inoculum size, then be that 28～32 ℃, fermentation raw material thickness are the 70～96h that ferments under the condition of 10～20cm at leavening temperature, namely obtain actinomycetes fertilizer.
The present invention utilizes the biogas fermentation waste---and natural pond slag, natural pond liquid replace original artificial synthetic medium as fermentation substrate, reduced and cultivated actinomycetic cost, in culturing process, by actinomycetes are cultivated, the high natural pond liquid of COD content is used, avoided direct discharging pollution on the environment.
Embodiment
Embodiment one: a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer of present embodiment, carry out according to the following steps: one, test tube is cultivated seed culture: a, preparation test-tube culture medium: take by weighing 2～2.5 parts of starch, 0.1～0.2 part in saltpetre, 0.03～0.05 part of potassium primary phosphate, 0.001～0.002 part in sal epsom, 2～2.2 parts in agar and 95～96 parts of water by mass fraction and mix, then carry out high-temperature sterilization, namely obtain test-tube culture medium; B, actinomycetes being inoculated in the test-tube culture medium, is that the inclined-plane leaves standstill cultivates 48～96h under 28～32 ℃ the condition in culture temperature, then use 1mL sterilized water wash-out test tube slant culture after, obtain elutriant; Two, solid spore seed culture: c, preparation seed culture medium: be 1 with millet in mass ratio with water: the ratio of (1～2) is mixed, and then immersion 20～24h carries out high-temperature sterilization, namely obtains seed culture medium; D, the elutriant in the step 1 being inoculated in the seed culture medium, is that shake-flask culture 72～120h obtains actinospore bacterium liquid under 28～32 ℃ the condition in culture temperature; The preparation of three, fermentation culture: e, fermentation raw material: take by weighing 70～90 parts of natural pond slags, 10～30 parts of wheat bran skins and CaCO by mass fraction
30.5～2 parts mix, and namely obtain fermentation raw material, the water content of regulating fermentation raw material with natural pond liquid is 40～45%, then carries out high-temperature sterilization; F, be in 0.5～5% fermentation raw material that is inoculated in after the sterilization with the actinospore bacterium liquid in the step 2 by inoculum size, then be that 28～32 ℃, fermentation raw material thickness are the 70～96h that ferments under the condition of 10～20cm at leavening temperature, namely obtain actinomycetes fertilizer.
The ratio that inoculum size in the present embodiment refers to move into the volume of seed liquor and inoculates rear nutrient solution volume.
Present embodiment is utilized the biogas fermentation waste---and natural pond slag, natural pond liquid replace original artificial synthetic medium as fermentation substrate, reduced and cultivated actinomycetic cost, in culturing process, by actinomycetes are cultivated, the high natural pond liquid of COD content is used, avoided direct discharging pollution on the environment.
Embodiment two: present embodiment and embodiment one are different is taking by weighing 2.1 parts of 2.2 parts of starch, 0.15 part in saltpetre, 0.04 part of potassium primary phosphate, 0.0015 part in sal epsom, agar by mass fraction and mix with 95.5 parts of water described in the step 1.Other steps are identical with embodiment one with parameter.
Embodiment three: present embodiment is different from embodiment one or two is the high-temperature sterilization described in the step 1 for being the 30min that sterilizes under 120 ℃ the condition in temperature.Other steps are identical with embodiment one or two with parameter.
Embodiment four: present embodiment is different from one of embodiment one to three is described in the step 2 to be that 1: 1.5 ratio is mixed in mass ratio with millet and water, soaks 24h.Other steps are identical with one of embodiment one to three with parameter.
Embodiment five: present embodiment is different from one of embodiment one to four is the high-temperature sterilization described in the step 2 for being the 20min that sterilizes under 120 ℃ the condition in temperature.Other steps are identical with one of embodiment one to four with parameter.
Embodiment six: present embodiment is different from one of embodiment one to five is that the preparation method of the natural pond slag described in the step 3 and natural pond liquid is high temperature biogas fermentation method, concrete grammar is: take by weighing 60 parts of 40 parts of fresh cow dungs, water by mass fraction and mix, 54 ℃ of cultivations of anaerobism 14 days are cultivated residuum and are natural pond slag and natural pond liquid.Other steps are identical with one of embodiment one to five with parameter.
Embodiment seven: present embodiment is different from one of embodiment one to six is the high-temperature sterilization described in the step 3 for being the 40min that sterilizes under 120 ℃ the condition in temperature.Other steps are identical with one of embodiment one to six with parameter.
Embodiment eight: present embodiment is different from one of embodiment one to seven is to take by weighing 80 parts of natural pond slags, 20 parts of wheat bran skins and CaCO by mass fraction described in the step 3
31.5 part mixes, and namely obtains fermentation raw material.Other steps are identical with one of embodiment one to seven with parameter.
Embodiment nine: what present embodiment was different from one of embodiment one to eight is that the leavening temperature described in the step 3 is 30 ℃.Other steps are identical with one of embodiment one to eight with parameter.
Embodiment ten: what present embodiment was different from one of embodiment one to nine is that the fermentation raw material thickness described in the step 3 is 15cm.Other steps are identical with one of embodiment one to nine with parameter.
By following verification experimental verification beneficial effect of the present invention:
Test 1: a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer of this test, carry out according to the following steps: one, test tube is cultivated seed culture: a, preparation test-tube culture medium: take by weighing 2.2 parts of starch, 0.15 part in saltpetre, 0.04 part of potassium primary phosphate, 0.0015 part in sal epsom, 2.1 parts in agar and 95.5 parts of water by mass fraction and mix, then the 30min that sterilizes under temperature is 120 ℃ condition namely obtains test-tube culture medium; B, actinomycetes being inoculated in the test-tube culture medium, is that the inclined-plane leaves standstill cultivates 72h under 30 ℃ the condition in culture temperature, then use 1mL sterilized water wash-out test tube slant culture after, obtain elutriant; Two, solid spore seed culture: a, preparation seed culture medium: be that 1: 1.5 ratio is mixed in mass ratio with millet and water, immersion 22h, the 20min that then sterilizes under temperature is 120 ℃ condition namely obtains seed culture medium; B, the elutriant in the step 1 being inoculated in the seed culture medium, is that shake-flask culture 96h obtains actinospore bacterium liquid under 30 ℃ the condition in culture temperature; The preparation of three, fermentation culture: a, fermentation raw material: take by weighing 80 parts of natural pond slags, 20 parts of wheat bran skins and CaCO by mass fraction
31.5 part mix, namely obtain fermentation raw material, be 43% with the water content of natural pond liquid adjusting fermentation raw material, 40min then sterilizes under temperature is 120 ℃ condition; B, being 3% to be inoculated in the fermentation raw material after the sterilization with the actinospore bacterium liquid in the step 2 by inoculum size, is that 30 ℃, fermentation raw material thickness are the 90h that ferments under the condition of 15cm at leavening temperature then, namely obtains actinomycetes fertilizer.Wherein the preparation method of the natural pond slag described in the step 3 and natural pond liquid is: high temperature biogas fermentation method: take by weighing 60 parts of 40 parts of fresh cow dungs, water by mass fraction and mix, 54 ℃ of anaerobism were cultivated 14 days, cultivated residuum and were natural pond slag, natural pond liquid.
Actinomycetes in this test are streptomyces microflavus GIM5406Streptomycesjingyangesis, buy from Shanghai north promise bio tech ltd.
This test utilizes the biogas fermentation waste---and natural pond slag, natural pond liquid replace original artificial synthetic medium as fermentation substrate, reduced and cultivated actinomycetic cost, in culturing process, by actinomycetes are cultivated, the high natural pond liquid of COD content is used, avoided direct discharging pollution on the environment.Application gradient dilution the method for plate culture count calculates in every gram actinomycetes fertilizer and contains spore alive more than 50,000,000,000.
Claims (10)
1. method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer, it is characterized in that utilizing the method for natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer to carry out according to the following steps: one, test tube is cultivated seed culture: a, preparation test-tube culture medium: take by weighing 2～2.5 parts of starch, 0.1～0.2 part in saltpetre, 0.03～0.05 part of potassium primary phosphate, 0.001～0.002 part in sal epsom, 2～2.2 parts in agar and 95～96 parts of water by mass fraction and mix, then carry out high-temperature sterilization, namely obtain test-tube culture medium; B, actinomycetes being inoculated in the test-tube culture medium, is that the inclined-plane leaves standstill cultivates 48～96h under 28～32 ℃ the condition in culture temperature, then use 1mL sterilized water wash-out test tube slant culture after, obtain elutriant; Two, solid spore seed culture: c, preparation seed culture medium: be 1 with millet in mass ratio with water: the ratio of (1～2) is mixed, and then immersion 20～24h carries out high-temperature sterilization, namely obtains seed culture medium; D, the elutriant in the step 1 being inoculated in the seed culture medium, is that shake-flask culture 72～120h obtains actinospore bacterium liquid under 28～32 ℃ the condition in culture temperature; The preparation of three, fermentation culture: e, fermentation raw material: take by weighing 70～90 parts of natural pond slags, 10～30 parts of wheat bran skins and CaCO by mass fraction
30.5～2 parts mix, and namely obtain fermentation raw material, the water content of regulating fermentation raw material with natural pond liquid is 40～45%, then carries out high-temperature sterilization; F, be in 0.5～5% fermentation raw material that is inoculated in after the sterilization with the actinospore bacterium liquid in the step 2 by inoculum size, then be that 28～32 ℃, fermentation raw material thickness are the 70～96h that ferments under the condition of 10～20cm at leavening temperature, namely obtain actinomycetes fertilizer.
2. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that taking by weighing 2.1 parts of 2.2 parts of starch, 0.15 part in saltpetre, 0.04 part of potassium primary phosphate, 0.0015 part in sal epsom, agar by mass fraction and mix with 95.5 parts of water described in the step 1.
3. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that the high-temperature sterilization described in the step 1 is in temperature being the 30min that sterilizes under 120 ℃ the condition.
4. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that described in the step 2 being that 1: 1.5 ratio is mixed in mass ratio with millet and water, soaks 24h.
5. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that the high-temperature sterilization described in the step 2 is in temperature being the 20min that sterilizes under 120 ℃ the condition.
6. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1, the preparation method who it is characterized in that the natural pond slag described in the step 3 and natural pond liquid is high temperature biogas fermentation method, concrete grammar is: take by weighing 60 parts of 40 parts of fresh cow dungs, water by mass fraction and mix, 54 ℃ of cultivations of anaerobism 14 days are cultivated residuum and are natural pond slag, natural pond liquid.
7. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that the high-temperature sterilization described in the step 3 is in temperature being the 40min that sterilizes under 120 ℃ the condition.
8. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that taking by weighing 80 parts of natural pond slags, 20 parts of wheat bran skins and CaCO by mass fraction described in the step 3
31.5 part mixes, and namely obtains fermentation raw material.
9. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that the leavening temperature described in the step 3 is 30 ℃.
10. a kind of method of utilizing natural pond slag, natural pond liquid-solid body fermentative production actinomycetes fertilizer according to claim 1 is characterized in that the fermentation raw material thickness described in the step 3 is 15cm.
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| CN103204615A (en) * | 2013-04-27 | 2013-07-17 | 罗炳 | Fermentation process of culturing excrement |
| CN103265154A (en) * | 2013-04-27 | 2013-08-28 | 罗炳 | Resource utilization and comprehensive treatment process for sewage and manure generated during cultivation |
| CN105254356A (en) * | 2015-09-23 | 2016-01-20 | 西北农林科技大学 | Bioorganic fertilizer used for planting organic vegetables, and preparation method thereof |
| CN107006331A (en) * | 2017-06-04 | 2017-08-04 | 湖南七纬科技有限公司 | The stereo plantation method that a kind of Cranberry is interplanted with Chinese yew |
| CN107114415A (en) * | 2017-06-04 | 2017-09-01 | 湖南七纬科技有限公司 | Matching used conditioning agent and growth accelerator when Chinese yew is interplanted in vineyard |
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| CN112219639A (en) * | 2020-10-15 | 2021-01-15 | 黑龙江省科学院微生物研究所 | Preparation method of decomposed biogas residues, culture medium and application of culture medium |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107189785A (en) * | 2017-06-04 | 2017-09-22 | 湖南七纬科技有限公司 | The conditioning agent that Cranberry uses when being interplanted with Chinese yew |
| CN107182693A (en) * | 2017-06-04 | 2017-09-22 | 湖南七纬科技有限公司 | A kind of method that Chinese yew is interplanted in vineyard |
| CN112219639A (en) * | 2020-10-15 | 2021-01-15 | 黑龙江省科学院微生物研究所 | Preparation method of decomposed biogas residues, culture medium and application of culture medium |
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