CN106434430A - Compound microbial agent for degrading antibiotic and pesticide residues as well as preparation and application thereof - Google Patents
Compound microbial agent for degrading antibiotic and pesticide residues as well as preparation and application thereof Download PDFInfo
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- CN106434430A CN106434430A CN201610807117.XA CN201610807117A CN106434430A CN 106434430 A CN106434430 A CN 106434430A CN 201610807117 A CN201610807117 A CN 201610807117A CN 106434430 A CN106434430 A CN 106434430A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 230000000593 degrading effect Effects 0.000 title claims abstract description 26
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- 239000003795 chemical substances by application Substances 0.000 title claims abstract description 11
- 230000000813 microbial effect Effects 0.000 title abstract description 16
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
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- Tropical Medicine & Parasitology (AREA)
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- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a compound microbial agent for degrading antibiotic and pesticide residues as well as preparation and application thereof, and belongs to the field of biotechnology and environmental protection. Multi-thallus compound microbial powder is prepared from the compound microbial agent according to the weight percentage of living microbes to the total amount of compound microbial powder as follows: 10%-15% of bacillus subtilis, 10%-15% of aspergillus niger, 10%-15% of bacillus mucilaginosus, 10%-15% of enterococcus faecalis, 8%-12% of bacillus licheniformis, 8%-12% of bacillus megaterium, 8%-12% of pseudomonas fluorescens, 5%-8% of lactobacillus plantarum, 5%-8% of bacillus polymyxin and 6%-8% of streptococcus thermophiles. The compound microbial agent has the effects of degrading antibiotic and pesticide residues, fermenting and composting organic matter, acting as functional fertilizer and repairing the environment, and can solve the problems of secondary pollution caused by antibiotic residues in culture feces and resource utilization of organic waste and realizes biodegradation of the antibiotic and pesticide residues in soil when applied to the agricultural ecological environment, thereby being of great value and practical significance in restoration of agricultural ecological environment and protection of human health.
Description
Technical field
The invention belongs to biotechnology and field of environment protection, it is related to biofermentation and processes cultivation manure contamination and organic
Utilization of waste as resource, microbial degradation antibiotic and pesticide residues carry out agroecological environment reparation, and in particular to a kind of
Degraded antibiotic and pesticide residues complex micro organism fungicide and its preparation and application.
Background technology
With scale and the batch production of China's aquaculture, in breeding process, Animal diseases are multiple, are to prevent and treat Animal diseases,
Various chemicalses, vaccine, antibiotic etc. are widely used during animal cultivation, cause medicine to be directly entered animal body
Interior, most of medicine passes through feces urine drains, forms the antibacterial medicine residue of high concentration in feces urine.
According to existing aquaculture treatment for cow manure technology, two kinds are divided into the processing mode of livestock and poultry feces Excreta：The first
It is that nature pile fermentation forms compost, the defect of this mode is cycle length, environmental pollution is serious, fertilizer efficiency is low, easily form two
Secondary pollution；The second way is that microbe leaven is seeded in Excreta, will make fertilizer after its fermentation maturity.Second
Although kind of a processing mode can achieve organic matter also field by deodorization of becoming thoroughly decomposed, the antibiotic for remaining in Organic substance is changed with other
Learn medicine not being decomposed, the antibiotic that the fertilizer for obtaining in this way is not degraded after applying farmland and chemicalses
Residue can still destroy beneficial microbial colonies balance in soil so that plant disease-resistant insect pest ability is weakened, and is Agro-ecology
Potential threat in environment and pollution.
In modern agriculture plantation, it is controlling crop diseases and insect pests, yield is improved, fertilizer and pesticide is generally excessively used.A lot
Pesticide application is to after farmland, and only 5% or so pesticide reaches target pest, and 95% pesticide remain on water body, soil and
In Agro-ecological System, it can enter eventually into biology in vivo by the enrichment of food chain, and the body for having a strong impact on the mankind is good for
Health.These pollutions are solved, maximally effective method is exactly to be degraded using microorganism.But existing agricultural composite microbial bacteria
Agent is broadly divided into two types by its effect：One kind is fermentation process livestock and poultry feces and organic materials, and after becoming thoroughly decomposed, Fertilizer Transformed is utilized；
One kind is used as functional living being fertilizer or biological pesticide.The complex micro organism fungicide of both types, acts only on
Fermentation maturity function, microorganism fixed nitrogen, Soluble phosphorus, potassium decomposing and part biological prophylactic-therapeutic effect to Organic substance, in livestock and poultry feces
Chemicalses cannot be degraded with pesticide residues in antibiotic remainss and agroecological environment.According to existing agricultural microbial bacteria
The technology of preparing of agent and product efficacy, hence it is evident that there is technological deficiency, i.e., can not solve agroecological environment Pesticides and antibiotic
Residual contamination.Current, be excessively used long-term with pesticide of chemical fertilizer has resulted in that soil beneficial microbe group is unbalance, soil plate
The lasting feeder capability of knot, soil weakens, pest and disease damage is occurred frequently, and agroecological environment bearing capacity is reached capacity, therefore, needs life badly
State is repaired and builds new ecological balance.
Content of the invention
The purpose of the present invention is the above-mentioned deficiency for prior art, provides a kind of degradable antibiotic and pesticide residues
Complex micro organism fungicide, the complex micro organism fungicide is prepared by microbial activity material, and specifically one kind is by bacillus subtilis
Bacterium, Bacillus licheniformis, aspergillus niger, colloid bacillus cereus, streptococcus thermophiluss, enterococcus faecalis, Lactobacillus plantarum, many viscous spore bars
The complex microorganism system that bacterium, bacillus megaterium, 10 kinds of microorganisms of pseudomonas fluorescens are constituted；The complex micro organism fungicide be according to
The different functionalities enzyme for producing according to the microorganism in complex microorganism system is prepared, with degraded antibiotic and pesticide residues,
The several functions such as organic matter fermentation becomes thoroughly decomposed, multi-Functional Fertilizers and repairing environment.
Technical scheme is as follows：The complex micro organism fungicide of a kind of degrading pesticide and antibiotic remainss, is to pass through
Bacterial screening and training systern, actication of culture, amplification culture, dehydrate step, respectively obtain bacillus subtilises,
Clothing bacillus cereuss, aspergillus niger, colloid bacillus cereus, streptococcus thermophiluss, enterococcus faecalis, Lactobacillus plantarum, bacillus polymyxa, huge
Bacterium anthracoides, the thalline dry powder of 10 kinds of strains of pseudomonas fluorescens, the thalline dry powder blend of 10 kinds of strains is uniform, obtain many
Thalline composite bacterium powder, as complex micro organism fungicide.
The degrading pesticide of the present invention and the complex micro organism fungicide of antibiotic remainss, its preparation method specifically includes following step
Suddenly：
（1）By bacillus subtilises, Bacillus licheniformis, aspergillus niger, colloid bacillus cereus, thermophilic coccus, enterococcus faecalis, plant
Lactobacilluss, bacillus polymyxa, bacillus megaterium, 10 kinds of bacterial strains of pseudomonas fluorescens carry out screening and the culture of bacterial strain respectively
Condition optimizing, obtains purification bacterial strain；Purification bacterial strain is carried out High Density Cultivation respectively, is inoculated into activation culture in shaking flask, will live
Change cultured strain be inoculated into fermentation tank be enlarged culture；After amplification culture terminates, list that amplification culture respectively is obtained
The zymocyte liquid of strain obtains the thalline dry powder of above-mentioned 10 kinds of strains through dehydrate；
（2）The thalline dry powder of 10 kinds of bacterial strains is uniformly mixed, obtains many thalline composite bacterium powder；In many thalline composite bacterium powder,
In each strain thalline dry powder live body bacterium number account for composite bacterium powder total viable count percentage by weight as follows：Bacillus subtilises 25-
30%th, Bacillus licheniformis 5-8%, aspergillus niger 8-10%, colloid bacillus cereus 12-15%, thermophilic coccus 2-6%, enterococcus faecalis 10-
13%th, Lactobacillus plantarum 16-22%, bacillus polymyxa 9-11%, bacillus megaterium 2-5%, pseudomonas fluorescens 15-25%；Institute
State in many thalline composite bacterium powder, effective active bacterial content is hundred million cfu/g of 200-500, moisture 5%；Compound in many thalline
Mycopowder is passed through filtrated air packaging, obtains final product complex micro organism fungicide.
Preferably, comprising the following steps that for bacillus subtilises thalline dry powder is prepared by bacillus subtilis strain：
（1）The screening of bacillus subtilis strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.015-0.02g/L, complementary element：DDT
0.1-0.2g/L, pH 7.0-7.2, by MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, Sodium Chloride, glucose, DDT, agar are dissolved in
In 1000mL distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basal medium
When 45 DEG C ~ 50 DEG C are cooled to, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis culture still in melt and dissolved state
Base mixes, and is then returned in sterilizing flat board, culture medium thickness at least high 5mm on flat board, and the final pH of culture medium is in 7.2-7.5
（When temperature is 25 DEG C）, the solid medium flat board containing DDT is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus subtilises that ampoul tube is preserved make bacterium solution, coat the training of the solid containing DDT
On foster base flat board, shading culture 2-3 days under the conditions of being placed in 35 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing DDT, be placed in 35 DEG C -38 DEG C, 180-200r/
Under min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of DDT concentration, 37-39 DEG C
Lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of DDT；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 2-3%
Inoculum concentration, be transferred in the fluid medium containing DDT, 35 DEG C -37 DEG C, 220-250r/min shaken cultivation 1-3 days, finally
With degradation rate of the high effective liquid chromatography for measuring bacterial strain to DDT；
7）Screening is obtained to DDT degradation rate highest bacillus subtilises, is preserved using test tube slant as purification bacterial strain, is placed in
4 DEG C of storages of refrigerator；
（2）Bacillus subtilis strain is activated：By step 7）Bacillus subtilises that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Being inoculated in activation culture shaking flask, with shaking flask rotating speed as 160-200r/min, 26h-32h is cultivated at a temperature of 35-37 DEG C, obtain
Liquid fermentation seed；The activation culture based formulas of above-mentioned activation culture are：Peptone 9.25-11.25g/L, Carnis Bovis seu Bubali cream 1.25-
1.3g/L、NaCl32.5-5.5g/L, glucose 22-25g/L, nutritional solution pH7.5-7.8；
（3）Bacillus subtilises amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 2-5%；The culture medium prescription of amplification culture is：Semen Maydis powder 18-20g/L,
Semen Glycines powder 20-22g/L, fish flour 5-6g/L, K2HPO41.2-1.5g/L、MgSO4 .7H2O 0.15-0.2g/L, Calcium Carbonate 6.5-
8.0g/L, ammonium sulfate 1.5-1.65g/L；In fermentation tank, temperature control is at 37-40 DEG C, and ferment pH7-8, and mixing speed is 200-
250r/min, fermentation time 28-32h；When the bacillus subtilis bacterial content in fermentation tank reaches 1010During individual/ml, will ferment
Bacillus subtilises bacterium solution drying dehydration be prepared into bacillus subtilises thalline dry powder.
Further, it is as follows that fermentation condition is best suitable for during the amplification culture：38.5 DEG C of fermentation jar temperature, pH7.0-7.5,
Fermentation time 30h.
According to the bacillus subtilises that above-mentioned steps are obtained, its characteristic with function is：Multiple antibiotics and raw egg can be produced
The white several enzymes of enzyme, α-amylase, cellulase, 1,4 beta-glucanase, phytase, pectase, xylanase etc. ten, anti-with wide spectrum
The active and extremely strong anti-adversity ability of bacterium；The sporinite of bacillus subtilises can successfully be converted into trophosome and to occupy advantage long
Phase survives and continues breeding and plays a role；Crop disease-resistant, cold-resistant, drought resisting energy can be improved, increases soil nutrient, improved soil knot
Structure, raising chemical fertilizer utilization ratio, promote the organic matter decomposition in soil to become humus, stimulate plant growth；Can fixed nitrogen, phosphorus decomposing, solution
Potassium, improves utilization rate of fertilizer；Similar cell mitogen, the material of plant growth activating element can be produced, and decomposes heavy metal, pesticide
Residual；Reduces cost, the yield that increases, raising income.
Preferably, by Bacillus licheniformis（Bacillus licheniformis）Bacterial strain prepares Bacillus licheniformis body
The comprising the following steps that of dry powder：
（1）The screening of lichem bacillus strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：MnSO40.5-1.0g/L, peptone 8-10g/L, cattle
Meat extract 2-3g/L, NaCl 3-5g/L, glucose 1.5-2.5g/L, agar 15-20g/L, complementary element：Bayer 71628 0.15-
0.25g/L, pH7.0-7.2, by MnSO4, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, Bayer 71628, agar be dissolved in 1000mL steaming
In distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled to 45
DEG C ~ 50 DEG C when, add and be dissolved in the complementary element of 2mL sterile purified water, mix with the basal medium still in melt and dissolved state, incline
Note in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium is in 7.0-7.2（When temperature is 25 DEG C）, must contain
There is the solid medium flat board of Bayer 71628, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Bacillus licheniformis that ampoul tube is preserved make bacterium solution, coat consolidating containing Bayer 71628
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 30 DEG C -32 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing Bayer 71628, be placed in 30 DEG C -32 DEG C, 160-
Under 180r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of methylamine phosphorus concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate is as capable of the bacterial strain of degrading methamidophos；
6）By step 5）It is capable of the bacterial strain of degrading methamidophos, is transferred on activation medium respectively and is activated；Then will activation
Bacterium solution press the inoculum concentration of 3%-5%, be transferred in the fluid medium containing Bayer 71628,30 DEG C -32 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to Bayer 71628；
7）Screening is obtained to methamidophos degradation rate highest lichem bacillus strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus licheniformis strain is activated：By step 7）Bacillus licheniformis that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Slant strains are inoculated in activation culture shaking flask, with shaking flask rotating speed 160r/min, cultivate 16h-20h, obtain at a temperature of 28-30 DEG C
Obtain liquid fermentation seed；The activation culture based formulas of the activation culture are：Peptone 9.25-10.0g/L, Carnis Bovis seu Bubali cream 4.5-
5g/L, NaCl 4.8-5.0g/L, glucose 1.5g-2.0/L, nutritional solution pH6.5-7.2；
（3）Bacillus licheniformis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 3-5%；Amplification culture based formulas are：Semen Maydis powder 15-18g/L, Semen Glycines powder
22-25g/L、K2HPO40.08-0.20g/L、MgSO4 .7H2O 0.005-0.01g/L, yeast powder 0.1-0.15g/L；Fermentation temperature
25-40 DEG C, ferment pH value 6-8, and it is 8-10%, fermentation time 18-28h that mixing speed is 200-300r/min, air mass flow ratio；
When the Bacillus licheniformis content in fermentation tank reaches 1010During individual/ml, by the Bacillus licheniformis liquid drying for fermenting
Dehydration is prepared into Bacillus licheniformis soma powder.
Further, it is as follows that fermentation condition is best suitable for during the amplification culture：29.5 DEG C of fermentation temperature, pH value 6.5-7.5,
Fermentation time 25h.
Bacillus licheniformis are obtained according to above-mentioned steps, its characteristic with function is：Various active enzyme can be produced, is albumen
Enzyme, amylase, lipase, pectase, glucanase, cellulase etc., the mechanism of its Biocontrol Effect is to compete, antagonism and induction
Based on plant resistance to environment stress；The antagonistic substance of generation mainly has antibiotic, bacteriocin, cell wall degradation enzyme and other antibacterial proteins,
To antibiotic and residual pesticide, there is certain degradation function；It is discharged in agricultural environment, soil microenvironment can be improved, make soil
Dominant population in earth is stable to keep on top；After being manured into soil, 100 times of rapid breeding becomes dominant bacteria, controls root
The nutrition on border and other resources, cause source of disease bacterium to lose vivosphere and condition to a great extent；Make plant related organization thin
Sclerosis, fibrosiss, degree of lignification are improved, and form the double silicon layers of cutin outside epidermal area, are formed together and are prevented pathogenic bacteria from being invaded
The barrier that attacks；Can improve the supply of nitrogen, phosphorus decomposing, potassium decomposing, the absorbance of Soil Phosphorus and potassium can be effectively improved.
Preferably, comprising the following steps that for aspergillus strain dry powder is prepared by Aspergillus niger strain：
（1）The screening of Aspergillus niger strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 5-10g/L, glucose 20-25g/L, sulfur
Sour magnesium 0.5-1.0g/L, yeast extract powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1.0-2.0g/L, agar 15-20g/L, mend
Fill composition：Glyphosate 0.15-0.25g/L, N,N'-dimethyl-.gamma..gamma.'-dipyridylium 0.2-0.5g/L, oxytetracycline 0.1-0.2g/L, by peptone, glucose,
Magnesium sulfate, yeast extract powder, dipotassium hydrogen phosphate, agar are dissolved in 1000mL distilled water, are subsequently adding 10mL glycerol, heating
Boil molten, and in 121 DEG C of high pressure steam sterilization 15min, basal medium is obtained, when basal medium is cooled to 45 DEG C ~ 50 DEG C, plus
Enter to be dissolved in the complementary element glyphosate of 2mL sterile purified water, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, oxytetracycline, with the basis culture still in melt and dissolved state
Base mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 6.5-6.8（Temperature is 25
DEG C when）, the solid medium flat board containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Aspergillus niger that ampoul tube is preserved makes bacterium solution, coats containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
On solid medium flat board, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated each bacterium colony, in the flat lining out of the solid medium containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, enters one
Step is separated and purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing in glyphosate, the fluid medium of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be placed in 30 DEG C -32 DEG C
Under 120-150r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, draws on the different inorganic salt solid medium flat board of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium concentration
Line, 30-32 DEG C of lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate as being capable of degradation of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Bacterial strain；
6）By step 5）Obtain can degradation of glyphosate, the bacterial strain of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be transferred on activation medium respectively and activated；
Then the bacterium solution of activation is pressed the inoculum concentration of 1-3%, is transferred to glyphosate, in N,N'-dimethyl-.gamma..gamma.'-dipyridylium fluid medium, 25 DEG C -28 DEG C,
160-180r/min vibration light culture 1-3 days, finally with high effective liquid chromatography for measuring bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Degradation rate；
7）Screening obtains the degradation rate highest Aspergillus niger bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, adopts as purification bacterial strain
Preserved with test tube slant, be placed in 4 DEG C of storages of refrigerator；
（2）Aspergillus niger strain is activated：By step 7）The aspergillus nigers for being placed in 4 DEG C of storages of refrigerator are inoculated into work in activation culture shaking flask
Change, activation culture based formulas are peptone 5.0-5.5g/L, glucose 10.0-15g/L, magnesium sulfate 0.5-0.8g/L, yeast
Powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1-3g/L, nutritional solution pH7.0-7.2 is leached, shaking flask rotating speed is 100-160r/min,
25h-28h is cultivated at a temperature of 28-35 DEG C, obtain liquid fermentation seed；
（3）Aspergillus niger amplification culture, dehydrate：By step（2）Obtain the fermentation that liquid fermentation seed is inoculated into amplification culture
Ferment in tank, be enlarged culture, inoculum concentration 8-10%；Amplification culture based formulas Rhizoma Solani tuber osi 35-40g/L, glucose 20-50g/
L, dipotassium hydrogen phosphate 5-8g/L, magnesium sulfate 1-1.5g/L, pH value are 7.0；Fermentation temperature is 35 DEG C, mixing speed 100-180r/
Min, fermentation time 25-28h, when the aspergillus niger content in fermentation tank reaches 1010During individual/ml, by the Aspergillus niger for fermenting
The dehydration of liquid drying is prepared into aspergillus strain dry powder.
Further, preferably fermentation time is 26h.
According to many aspergillus nigers that above-mentioned steps are obtained, its characteristic with function is：Can secretion generation amylase, saccharifying enzyme, lemon
Lemon acid, gluconic acid, gallic acid etc., with cracking larger molecular organicses and indissoluble inorganic matters ability, are easy to Crop profit
With improving Soil structure, strengthen soil fertility, improve the effect of crop yield；Under other microorganism Co metabolism effects, to hundred
The herbicides such as careless withered, glyphosate and chemical pesticide have certain Degradation.
Preferably, by colloid bacillus cereus（Bacillus mucilaginosus Krassilnikov）Bacterial strain prepares glue
The comprising the following steps that of matter bacillus cereuss thalline dry powder：
（1）The screening of colloid bacillus cereus bacterial strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 2-3g/L, ammonium sulfate 0.15-0.25g/L,
Yeast extract 0.1-0.g/L, KCl 0.01-0.02g/L, MgSO4 .7H2O 0.01-0.02g/L、NaH2PO40.01-0.02g/L、
CaCO30.1-0.2g/L, silica 1-2g/L, agar 10-15g/L, complementary element：Cypermethrin 0.1-0.3g/L,
CuSO4 .5H2O0.02-0.025g/L, initial pH 7.0-7.2；By glucose, ammonium sulfate, yeast extract, KCl, MgSO4 .7H2O、
NaH2PO4、CaCO3, silicon dioxide, CuSO4 .5H2O, agar are dissolved in 1000mL distilled water, heating boil molten, and at 121 DEG C
High pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL sterilizing
The complementary element cypermethrin of distilled water, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, training
Foster base thickness at least high 5mm, the final pH of culture medium is in 7.0-7.5（When temperature is 25 DEG C）, must have cypermethrin and copper sulfate
Solid medium flat board, be placed in dark place, in 3 DEG C ~ 6 DEG C preserve；
2）Inoculation and purification：The common colloid bacillus cereus that ampoul tube is preserved make bacterium solution, coat containing cypermethrin and
On the solid medium flat board of copper sulfate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of cypermethrin concentration, 32-
35 DEG C of lucifuge cultures 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of cypermethrin；
6）By step 5）Obtain degrading the bacterial strain of cypermethrin, is transferred on activation medium respectively and is activated；Then
The bacterium solution of activation is pressed the inoculum concentration of 2-5%, has been transferred in cypermethrin fluid medium, 37 DEG C -40 DEG C, 200-220r/
Min shaken cultivation 1-3 days, finally uses degradation rate of the high effective liquid chromatography for measuring bacterial strain to cypermethrin；
7）Screening obtains the degradation rate highest colloid bacillus cereus bacterial strain to cypermethrin, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Colloid bacillus cereus actication of culture：By step 7）Colloid bacillus cereus inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Starch 15-20g/L, dipotassium hydrogen phosphate g/L, ammonium sulfate 2-3g/
L, yeast extract 3.5-5.5g/L, sucrose 8.5-10g/L, magnesium sulfate 4.5-5g/L, Calcium Carbonate 2.5-3g/L, silicon dioxide 6.5-
8g/L, ferric chloride 0.05-lg/L, pH7.0-7.2, shaking flask rotating speed is to cultivate at a temperature of 30-35 DEG C
18-20h, obtains liquid fermentation seed；
（3）Colloid bacillus cereus amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 4-6%；The composition of fermentation medium is sucrose 2.5-6.5g/L, phosphorus
Sour hydrogen dipotassium 3-5.5g/L, yeast extract 3-6g/L, ammonium sulfate 2.5-5g/L, soy peptone 5-10g/L, magnesium sulfate 3-6/g/
L、pH7.2-7.5；Amplification culture condition：32 DEG C of fermentation temperature, pH7.2, mixing speed 180r/min-220r/min, during fermentation
Between 24h；When the colloid bacillus cereus content of fermentation tank reaches 1010During individual/ml, by the colloid bacillus cereus bacterium solution warp for fermenting
Drying and dehydrating is prepared into colloid bacillus cereus thalline dry powder.
According to many colloid bacillus cereus that above-mentioned steps are obtained, its characteristic with function is：Silicate and aluminum silicate can be decomposed
Potassium-bearing mineral in salt and other Ores, with Soluble phosphorus, Potassium release and fixed nitrogen function, while can have during growth and breeding
Machine acid, aminoacid, polysaccharide, hormone etc. are conducive to plant absorption and the material for utilizing；After breeding in soil, plant growing is secreted
Stimulin and multiple enzymes, to strengthen resistance of the crop to some diseases, can suppress the growth of other pathogen；In thalline ash
Potassium content more than 33%, endobacillary potassium can thalline death after separate out, can directly be absorbed by plant；Have
Activation chesson, the various middle trace element of decomposition, silico-calcium sulfur boron molybdenum zinc etc.；Produce multiple physiology such as gibberellins, heteroauxing
Active substance, makes plant growth stalwartness, strengthens the disease-resistant and anti-adversity ability of crop cold resistance drought resisting, improves crop yield and improves product
Quality；Profitable strain, the effectively generation of suppression soil-borne disease are formed in crop root；With certain Adsorption of Heavy Metals ability and with
Other microorganism co -metabolic degradation pesticide.
Preferably, comprising the following steps that for streptococcus thermophiluss thalline dry powder is prepared by strains of streptococcus thermophilus：
（1）The screening of strains of streptococcus thermophilus is carried out according to a conventional method with training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 2-3g/L, peptone 2-3g/
L, yeast extract 1-2g/L, Fructus Lycopersici esculenti juice 15-20g/L, glucose 2-4g/L, tween 0.5ml-1.0ml/L, Calcium Carbonate 1.0-2.0g/
L, agar 2-5g/L, pH6.5-7.0, take suitable quantity of water in pot according to ratio, heating, by Carnis Bovis seu Bubali cream, peptone, yeast extract, kind
Tomato juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour in the conical flask for added agar and add after medicine boiling in pot
Plug, wrapping, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator culture 24h, asepsis growth person
Can use；The culture medium using when can also make fluid medium without agar, for the shaken cultivation of mushroom；
The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common streptococcus thermophiluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, culture obtains bacterial strain；
6）By step 5）Each bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 5%-
8% inoculum concentration, is transferred in fluid medium, 37 DEG C, quiescent culture 1-2 days, determines the biological active matter produced by each bacterial strain
Matter content；
7）Screening obtains producing bioactive substance content highest strains of streptococcus thermophilus, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Streptococcus thermophilus strain is activated：By step 7）Streptococcus thermophiluss obtaining, being placed in 4 DEG C of storages of refrigerator are inoculated into work
Change activation culture in culture shaking flask, activation culture based formulas are：Tryptone 1-3.0g/L, fish peptone 2-3g/L, Carnis Bovis seu Bubali cream
2-3g/L, yeast extract 0.5-1.5g/L, magnesium sulfate 0.3-0. 5g/L, potassium dihydrogen phosphate 0.5-1.0g/L, Lactose 1-1.5g/L,
Nutritional solution pH 6.5-6.8, cultivates 24h-28h at a temperature of 35-40 DEG C, and mixing speed is 80-100r/min, obtains liquid and sends out
Ferment seed；
（3）Streptococcus thermophiluss amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 2-5%；Amplification culture based formulas：Glucose 1-3g/L, soy peptone 4-
6g/L, dipotassium hydrogen phosphate 2-3g/L, corn juice 40-50g/L, sodium acetate 1-2g/L, skimmed milk 10-15g/L, pH are 6.5-6.8；
38-45 DEG C of fermentation temperature, mixing speed is 100-120r/min, fermentation time 48-72h；When the streptococcus thermophiluss in fermentation tank
Content reaches 109During individual/ml, the streptococcus thermophiluss bacterium solution drying for fermenting dehydration is prepared into streptococcus thermophiluss thalline and is done
Powder.
According to many streptococcus thermophiluss that above-mentioned steps are obtained, its characteristic with function is：Can suppress harmful in pollution environment
Bacterium pathogen, such as clostridium difficile, Salmonella typhimurium, escherichia coli, staphylococcus aureus, monocyte hyperplasia Liszt
Bacterium, bacillus perfringens etc.；During fermentation animal wastes, can suppress or eliminate harmful pathogen quickly, shorten fermentation
Process, improves the ferment effect that becomes thoroughly decomposed；With some functional activities, extracellular polysaccharide, bacteriocin and vitamin is produced.
Preferably, by E. Faecium strains, comprising the following steps that for enterococcus faecalis thalline dry powder is prepared：
（1）The screening of E. Faecium strains and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 10-15g/L, maltose 20-25g/L, ferment
Female leaching powder 10-2g/L, sodium glycerophosphate 5-6g/L, Sodium Chloride 1-2g/L, Lactose 0.015-0.020g/L, agar 15-20, pH
Value 8.0-8.5, is separately separately added into complementary element in order in the medium：Erythromycin 0.05-0.15g/L, ciprofloxacin
0.01-0.02g/L, Sulfamethoxazole Compound 0.25-0.35g/L；By peptone, maltose, yeast extract, sodium glycerophosphate,
Sodium Chloride, Lactose, agar are dissolved in 1000mL distilled water, are subsequently adding 10mL glycerol, and heating is boiled molten and high at 121 DEG C
Pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL sterilizing steaming
The complementary element erythromycin 0.05-0.15g/L of distilled water, ciprofloxacin 0.01-0.02g/L, Sulfamethoxazole Compound 0.25-
0.35g/L, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, and culture medium thickness are at least high
5mm, the final pH of culture medium is in 7.5-8.0（When temperature is 25 DEG C）, containing erythromycin, ciprofloxacin, compound sulfonamide first dislike
Azoles solid medium flat board, is placed in dark place, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common enterococcus faecalis that ampoul tube is preserved make bacterium solution, coat husky containing erythromycin, ring third
On star, Sulfamethoxazole Compound solid medium flat board, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium
Fall；
3）By step 2）Isolated bacterium colony, containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound solid medium
Flat lining out, is separated and purification further；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound liquid culture
In base, 33 DEG C -35 DEG C are placed in, under 160-180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, solid in the different inorganic salt of erythromycin, ciprofloxacin, Sulfamethoxazole Compound concentration
Rule on body culture medium flat plate, 35 DEG C -38 DEG C lucifuge cultures 2-5 days；The bacterial strain that can grow on culture medium flat plate is energy
Enough degraded erythromycin, ciprofloxacin, the bacterial strain of Sulfamethoxazole Compound；
6）By step 5）Obtain degrading the bacterial strain of erythromycin, ciprofloxacin, Sulfamethoxazole Compound, is transferred to activation respectively
Activated in culture medium, then the bacterium solution of activation pressed the inoculum concentration of 3%-5%, be transferred to pH8.5-9.0 containing erythromycin,
In ciprofloxacin, Sulfamethoxazole Compound element fluid medium, at 38 DEG C -40 DEG C, 180-250r/min vibrates light culture 1-3
My god, finally with each bacterial strain of high effective liquid chromatography for measuring to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate；
7）Screening obtain to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate highest E. Faecium strains, as
Purification bacterial strain is preserved using test tube slant, is placed in 4 DEG C of storages of refrigerator；
（2）Enterococcus faecalis actication of culture：By step 7）The enterococcus faecalis for being placed in 4 DEG C of storages of refrigerator for obtaining are inoculated into activation culture
Activate in shaking flask, activation culture based formulas are：Glucose 4-5g/L, yeast powder 0.15-0.3g/L, peptone 0.5-1g/L, no
Water calcium chloride 0.002-0.006g/L, bitter salt 0.02-0.05g/L, dipotassium hydrogen phosphate 0.1-0.15g/L, seven hydrations
Manganese sulfate 0.002-0.005g/L, Calcium Carbonate 0.5-1g/L, nutritional solution pH7.5-8.0, shaking flask rotating speed is 180-200r/min,
30h-35h is cultivated at a temperature of 30-35 DEG C, obtain liquid fermentation seed；
（3）Enterococcus faecalis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into sending out for amplification culture
Ferment in fermentation tank, be enlarged culture, inoculum concentration 8-10%；Culture medium prescription is tryptone 10-15g/L, yeast powder 10-
15g/L, glucose 10-15g/L, anhydrous calcium chloride 0.04-0.06g/L, bitter salt 0.019-0.025g/L, phosphoric acid hydrogen
Dipotassium 0.04-0.06g/L, 0.04-0.06g/L of potassium dihydrogen phosphate, dibastic sodium phosphate 0.4-0.6g/L, Sodium Chloride 0.08-0.1g/
L, murphy juice 50-80g/L, Calcium Carbonate 1-3g/L, sodium acetate 10-15g/L, ammonium carbonate 2-3g/L, ammonium acetate 1-2g/L, half Guang ammonia
Acid hydrochloride 0.5-1g/L, pH value is 8.5-9；35-37 DEG C of fermentation temperature, mixing speed 180-220r/min, fermentation time 20-
32h, when the enterococcus faecalis content of fermentation tank reaches 1010During individual/ml, by the enterococcus faecalis bacterium solution drying for fermenting dehydration system
Standby become enterococcus faecalis thalline dry powder.
Further, the fermentation time is 25h.
According to many enterococcus faecalis that above-mentioned steps are obtained, its characteristic with function is：Can produce in growth and metabolic process
Beneficial nutrient substance, such as lactic acid, aminoacid, vitamin, enzyme and antibacterial substance；Enterococcus faecalis and bacillus cereuss and aspergillus niger shape
The alternate Co metabolism for becoming good is acted on, to penicillins, erythromycin, quinolones, nitrofurantoin, high concentration gentamycin and height
Concentration streptomycin and rifampicin have degradation capability；Enterococcus faecalis are aerobic and facultative anaerobic bacteria, in fermentation animal wastes process
In, energy fast deodorization, control harmful gass volatilization.
Preferably, comprising the following steps that for Lactobacillus plantarum thalline dry powder is prepared by lactobacillus plantarum strain：
（1）The screening of lactobacillus plantarum strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 1-2g/L, peptone 1-2g/
L, yeast extract 1-2g/L, 20-30g/L of Fructus Lycopersici esculenti juice, glucose 1-2g/L, tween 0.05ml-0.10ml/L, Calcium Carbonate 1.5-
2.0g/L, agar 2-5g/L, pH6.2-6.5, according to ratio according to the required suitable quantity of water that measures in pot, heating, weigh Carnis Bovis seu Bubali cream,
Peptone, yeast extract, Fructus Lycopersici esculenti juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour into and add after medicine boiling in pot
Jump a queue in the conical flask of agar, wrap up, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator trainings
Foster 24h, asepsis growth person can use；Culture medium using when can also make fluid medium without agar, for bacterium
The shaken cultivation of class；The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common plant lactobacilluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）The each bacterial strain for obtaining, is transferred on activation medium respectively and is activated, and then presses the bacterium solution of activation
The inoculum concentration of 3%-5%, is transferred in fluid medium, 37 DEG C, and quiescent culture 1-2 days finally determines the life produced by each bacterial strain
Active substances content；
7）Screening obtains producing bioactive substance content highest lactobacillus plantarum strain, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Lactobacillus plantarum actication of culture：By step 7）Lactobacillus plantarum obtaining, being placed in 4 DEG C of storages of refrigerator is inoculated into work
Change and activate in culture shaking flask, activation culture based formulas are：Peptone 10-12g/L, Carnis Bovis seu Bubali cream 10-15g/L, yeast extract 5-6.5g/
L、KH2PO42-3.5g/L, trisodium citrate 2-2.5g/L, sodium acetate 2-2.5g/L, glucose 20-28g/L, MgSO4 .7H2O
0.58-0.60g/L、MnSO4 .4H2O 0.25-0.3g/L, nutritional solution pH 6.2-6.5, cultivate 24h- at a temperature of 35-40 DEG C
28h, quiescent culture, obtain liquid fermentation seed；
（3）Lactobacillus plantarum amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 5-8%；Culture medium prescription is：Sucrose 25-30g/L, yeast extract 15-20g/
L、KH2PO48-10g/L, trisodium citrate 5-6g/L, sodium acetate 3-5g/L, MgSO4 .7H2O 0.8-1.2g/L、MnSO4 .4H2O
0.5-0.8g/L, pH 6.2-6.5；35-37 DEG C of fermentation temperature, fermentation time 24-28h, quiescent culture；Fermentation ends, will fermentation
Good Lactobacillus plantarum bacterium solution drying dehydration is prepared into Lactobacillus plantarum thalline dry powder.
Further, 36 DEG C of the fermentation temperature, fermentation time 25h.
According to many Lactobacillus plantarum that above-mentioned steps are obtained, its characteristic with function is：Substantial amounts of acid can be produced, make the pH in water
Value stabilization is not raised, and the acidic materials of its output can be degraded heavy metal, and Lactobacillus plantarum has that breeding is quick, vitality
By force, the features such as safety non-toxic；Can the multiple organic acid of secretion synthesis, enzyme, biological active substanceies.
Preferably, comprising the following steps that for bacillus polymyxa thalline dry powder is prepared by bacillus polymyxa bacterial strain：
（1）The screening of bacillus polymyxa bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-15g/L, yeast extract 5-8g/
L、NaCl 3-5g/L、KH2PO40.1-0.2g/L、MgSO4 .7H2O 0.05-0.1g/L、MnSO4 .7H2O 0.25-0.5g/L, fine jade
Fat 15-20g/L, pH7.5-8.0；By MnSO4 .7H2O, peptone, KH2PO4、NaCl、MgSO4 .7H2O, agar are dissolved in 1000mL
In distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled to
When 45 DEG C ~ 50 DEG C, it is poured in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium is in 7.5-8.0（Temperature
During for 25 DEG C）, solid medium flat board is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus polymyxa that ampoul tube is preserved makes bacterium solution, coats solid medium flat board
On, shading culture 2-3 days under the conditions of being placed in 37 DEG C -40 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single dominant bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be placed in 38 DEG C -40 DEG C, 160-180r/min vibrate bar
Under part, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 38-40 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 2-5%
Inoculum concentration, be transferred in fluid medium, 37 DEG C -40 DEG C, 180-200r/min shaken cultivation 1-3 days, finally determine each bacterium
The various bioactivators contents such as such as antibiotic, antagonist protein, phytohormone, enzyme, flocculant produced by strain；
7）Screening obtains producing bioactive substance content highest bacillus polymyxa bacterial strain, adopts test tube as purification bacterial strain
Inclined-plane is preserved, and is placed in 4 DEG C of storages of refrigerator；
（2）Bacillus polymyxa actication of culture：By step 7）Screening obtains, is placed in the bacillus polymyxa of 4 DEG C of storages of refrigerator and connects
Plant and activate in activation culture shaking flask, activation culture based formulas are：Peptone 8-10g/L, Carnis Bovis seu Bubali cream 3-5g/L, Sodium Chloride 3-
5g/L, glucose 10-12g/L, KH2PO40.2-0.3g/L、MgSO4 .7H2O 0.2-0.35g/L、NaCl 0.2-0.3g/L、
CaSO4 .2H2O 0.2-0.25g/L、CaCO35.0-6.5g/L, nutritional solution pH 7.0-8.0, shaking flask rotating speed 150-170r/min,
25h-30h is cultivated at a temperature of 37-40 DEG C, obtain liquid fermentation seed；
（3）Bacillus polymyxa amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 10-12%；Amplification culture based formulas are：Semen Maydis powder 20-25g/L, Semen Glycines powder
8-12g/L、K2HPO40.05-0.15g/L, Sodium Chloride 1.0-1.5g/L, Na2HP040.15-0.25g/L；Fermentation temperature 28-
35 DEG C, ferment pH7-8, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h；When the bacillus polymyxa of fermentation tank contains
Amount reaches 1010During individual/ml, the bacillus polymyxa bacterium solution drying for fermenting dehydration is prepared into bacillus polymyxa thalline
Dry powder.
Further, 32 DEG C of fermentation temperature, pH 7.0-7.5, fermentation time 22h.
According to the bacillus polymyxa that above-mentioned steps are obtained, its characteristic with function is：Such as antibiotic, antagonism egg can be produced
In vain, the various bioactivators such as phytohormone, enzyme, flocculant；Multiple animals and plants pathogenetic bacterias and funguses are had by has a broad antifungal spectrum
There is stronger antagonism；Fermentation performance is excellent, can produce multi-component cyclic peptide antibacterial active substance, while can produce anti-true
The active substance of bacterium, has the effect of biological pesticide and bio-bacterial manure concurrently；Paenibacillus polymyxa is that a kind of plant rhizosphere growth-promoting is thin
Bacterium, can be used as microbial manure, promotes plant growing, improves crop yield；Vegetative bacteria and fungoids soil can effectively be prevented and treated
Disease is passed, the polypeptide antibiotic substance that Paenibacillus polymyxa is produced has good antagonistic activity to plant pathogenic fungi, and
Nature comparison is stable, can be used for the Biological control of plurality of plant diseases.
Preferably, comprising the following steps that for bacillus megaterium thalline dry powder is prepared by bacillus megaterium bacterial strain：
（1）The screening of bacillus megaterium bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.025-0.035g/L, complementary element：To sulfur
Phosphorus 0.15-0.20g/L, pH7.5-8.0；By MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, parathion, agar are molten
Solution is in 1000mL distilled water, and heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basic training
When foster base is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis still in melt and dissolved state
Culture medium mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 7.5-8.0（Temperature
During for 25 DEG C）, the solid medium flat board containing parathion is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus megaterium that ampoul tube is preserved makes bacterium solution, coats consolidating containing parathion
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of parathion concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of parathion；
6）By step 5）Obtain degrading the bacterial strain of parathion, is transferred on activation medium respectively and is activated, then will
The bacterium solution of activation presses the inoculum concentration of 2-5%, has been transferred in parathion fluid medium, and 37 DEG C -40 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to parathion；
7）Screening obtains the degradation rate highest bacillus megaterium bacterial strain of parathion, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus megaterium actication of culture：By step 7）Bacillus megaterium inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Peptone 10-15g/L, Carnis Bovis seu Bubali cream 3-5g/L, NaCl6.5-
8.5g/L, glucose 2.5-3g/L, nutritional solution pH7.0-7.2, shaking flask rotating speed is to cultivate 190-210r/min, at 37-40 DEG C
22-28h, obtains liquid fermentation seed；
（3）Bacillus megaterium amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5-9%；The culture medium prescription of amplification culture is：Semen Maydis powder 15-18g/L,
Semen Glycines powder 25-30g/L, fish flour 5-6g/L, K2HPO40.05-0.15g/L, calcium chloride 5.5-6.5.0g/L, ammonium sulfate 2.0-2.5g/
L；36-40 DEG C of fermentation temperature, fermentation pH value issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss；
When the bacillus megaterium content in fermentation tank reaches 1010During individual/ml, by the bacillus megaterium bacterium solution drying for fermenting
Dehydration is prepared into bacillus megaterium thalline dry powder.
Further, 39 DEG C of the fermentation temperature, pH 7.0-7.2, fermentation time is 28h.
According to the bacillus megaterium that above-mentioned steps are obtained, its characteristic with function is：With extremely strong heat resistanceheat resistant, radioprotective,
Some special properties such as anti-chemicalses and hydrostatic pressure resistant；Compared with its easy preservation of nutrition somatic cell, resurrection rate height；Have well
In degraded soil, effect of organophosphors, has repair to the water body for polluting, and passes through symbiosis with other bacillus cereuss of nature
And Co metabolism, the antibiotic for remaining in environment of degrading and pesticide；Competing by colonizing, secreting antibiotic substance and nutrition in plant rhizosphere
Strive and antagonism pathogen.
Preferably, comprising the following steps that for pseudomonas fluorescens thalline dry powder is prepared by pseudomonas fluorescens strain：
（1）The screening of pseudomonas fluorescens strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-12g/L, K2SO410-12g/L、
MgCl21.4-1.6g/L、Fe2(SO4)30.5-1g/L, glycerol 10mL, 15 ~ 20g/L of agar, 1 L of distilled water, complementary element：
Penicillin 0.05-0.10g/L；By tryptone, K2SO4、MgCl2、Fe2(SO4)3, agar be dissolved in 1000mL distilled water,
It is subsequently adding 10mL glycerol, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, basal medium is obtained, treats basic culture
When base is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis training still in melt and dissolved state
Foster base mixing, is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 7.0-7.2（Temperature is
When 25 DEG C）, the solid medium containing penicillin is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common fluorescent pseudomonass that ampoul tube is preserved make bacterium solution, coat consolidating containing penicillin
On body culture medium flat plate, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing penicillin, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing penicillin, be placed in 25 DEG C -28 DEG C, 150-
Under 180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of penicillin concn, 25-28
DEG C lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of penicillin；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 1%-3%
Inoculum concentration, be transferred to containing in penicillin fluid medium, at 25 DEG C -28 DEG C, 160-180r/min vibrates light culture 1-3
My god, finally use degradation rate of the high effective liquid chromatography for measuring bacterial strain to penicillin；
7）Screening is obtained to penicillin degradation rate highest pseudomonas fluorescens strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Pseudomonas fluorescens actication of culture：By step 7）The pseudomonas fluorescens inclined-plane bacterium for being placed in 4 DEG C of storages of refrigerator for obtaining
Plant to be inoculated in activation culture shaking flask and activate, activation culture based formulas are：Carnis Bovis seu Bubali cream 2.55-3.0g/L, peptone 1.65-
2.2g/L, Sodium Chloride 0.18g-0.2g/L, yeast extract 5.25-6.0g/L, Fe2(SO4)31-1.5g/L, culture fluid pH7.0-
7.5, shaking flask rotating speed is to cultivate 18-22h at a temperature of 28.5-31 DEG C, obtains liquid fermentation seed；
（3）Pseudomonas fluorescens amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5.5 ~ 8.5%；The culture medium prescription of the amplification culture is：Maltose
20-35g/L, Semen Maydis powder 12-15g/L, yeast extract 4.5-4.62g/L, ammonium sulfate 5.2-5.8g/L, K2HPO40.55-0.85g/
L、MgSO4 .7H2O 0.035-0.055g/L、FeSO4 .7H2O 1.095-1.20g/L、CaCl20.075-1.0g/L、NaCl
0.15-0.25g/L；27 DEG C ~ 29 DEG C of fermentation temperature, fermentation tank 140 ~ 180r/min of mixing speed, ventilation 8 ~ 12%, during fermentation
Between 24 ~ 48h；When the Pseudomonas bacterial content in fermentation tank reaches 1010During individual/ml, by the pseudomonas fluorescens for fermenting
The dehydration of bacterium solution drying is prepared into pseudomonas fluorescens thalline dry powder.
According to the pseudomonas fluorescens that above-mentioned steps are obtained, its characteristic with function is：Have good degraded to make to penicillium sp
With with decomposition of cellulose, protein and combining Fe3+Ability；Chitinase, lysozyme, xylanase and extracellular can be produced
Chitosanase etc. is degraded to material, can also reduce the generation of plant disease while nutrition is provided to plant；Can carry out
Denitrification, decomposable asymmetric choice net fat.
Complex micro organism fungicide described in technical solution of the present invention can be used as organic pollution innocent treatment agent, and it is right to be applied to
Feces of livestock and poultry carries out the harmless treatment of compost fermentation, can also be to each type organic of agriculture waste, the organic original of industrial processes
The harmless treatment of the organic pollutions such as material, organic waste, organic sludge.
Complex micro organism fungicide described in technical solution of the present invention is alternatively arranged as soil conditioner etc., is applied in agricultural planting
Improved soil, Biological control, the agriculture border pollution of reduction, increase crop yield and raising crop product quality.
Complex micro organism fungicide described in technical solution of the present invention is alternatively arranged as agricultural environment renovation agent, is applied to agriculture of degrading
Medicine, process heavy metal pollution.
Common bacillus subtilises of the present invention, common Bacillus licheniformis, common aspergillus niger, common colloid spore
Bacillus, commonly thermophilic coccus, common enterococcus faecalis, common plant lactobacilluss, commonly common bacillus polymyxa, huge spore bar
Bacterium, 10 kinds of original strains of common fluorescent pseudomonass refer to without screening, purification, can be directly from Chinese Academy of Sciences's microorganism
The mechanism of the authorized by states such as institute common micro-organisms center obtains.
Drying and dehydrating of the present invention can dry conventional method using bacterium solution, preferably can adopt spray drying method, do
Mass content≤5% of the thalline dry powder moisture for obtaining after dry.
Complex micro organism fungicide described in technical solution of the present invention can be used for the know-why of degrading pesticide and antibiotic remainss
As follows：
（1）The present invention is to set up the ecological balance of microbiologic population using the intergrowth relation between microorganism, using microorganism altogether
Metabolism, supplements composite microbial bacteria and environment indigenous microorganisms symbiosis in agricultural environment, sets up new microbial ecological
System, by the antibacterial in microflora or funguses under some natural endowments and non-existent by synthetic insecticide, kill
The chemical substances such as microbial inoculum, herbicide are degraded；
（2）The present invention is to allow microorganism directly using pesticide or antibacterial as growth substrate, and in multiple-microorganism group
Under effect, pesticide and the structure of antibiotic residues is made to change, so as to cause its chemical and physical features to change, i.e.,
By pesticide or antibiotic residues are degraded to micromolecular compound from macromolecular compound, finally become H20 and C02, by its
Resolve into inorganic matters completely, realize being completely degraded, become nontoxic inorganic matters；
（3）The present invention be using microorganism enzymatic route and non-enzymatic route degrading pesticide.Enzymatic reaction is microorganism itself
Enzyme system gene containing the degradable pesticide, directly acts on pesticide by the effect such as oxidation, dehydrogenation, reduction, hydrolysis, synthesis,
Or, although the enzyme system without the pesticide of degrading, but under pesticide stress, there is restructuring or change in the gene of microorganism, produce
New degradation enzyme system；Non- enzymatic reaction refers to that microbial activitiess make the pH of environment change and cause degradation of pesticide, or produces
Cofactor or chemical substance participate in the conversion of pesticide.
The present invention having the beneficial effect that with respect to prior art：
（1）The complex micro organism fungicide for being prepared using technical solution of the present invention is by feces of livestock and poultry, agricultural crop straw, organic waste etc.
Agriculture pollutants carries out compost maturity harmless treatment, can will remain in the agriculturals such as feces of livestock and poultry, agricultural crop straw, organic waste
Antibiotic in pollutant and drug degradation, organic matter after harmless treatment and beneficial microbe may be directly applied to dirty
The environment of dye, effectively eliminates pollution by pesticides；
（2）Using technical solution of the present invention by complex micro organism fungicide directly as bio-feritlizer or agroecological environment renovation agent
Utilize, the beneficial microbe in complex micro organism fungicide can be discharged and add in environment, primary beneficial micro- with environment
Biology is collectively forming stable group's system, by alternate and the Co metabolism of microorganism, realizes agricultural environment Pesticide Residues
Degradable；
（3）The present invention compared with traditional physics, chemical method, with put into low, regulation effect substantially, do not produce side effect,
Can recover and build the feature of ecological environment, it is possible to provide a kind of method of the environmentally friendly removal pollutant of low cost.
Specific embodiment
Below by embodiment, the present invention is described in further details, these embodiments are only used for the present invention is described, and
Do not limit the scope of the invention.
Strain source of the present invention is in Institute of Microorganism, Academia Sinica's common micro-organisms center（CGMCC）,
Its numbering is：Bacillus subtilises CGMCC1.1086, Bacillus licheniformis CGMCC1.0265, aspergillus niger CGMCC3.0316, glue
Matter bacillus cereuss CGMCC1.0910, streptococcus thermophiluss CGMCC1.1855, enterococcus faecalis CGMCC1.2135, Lactobacillus plantarum
CGMCC1.0557, bacillus polymyxa CGMCC1.0548, bacillus megaterium CGMCC1.0217, pseudomonas fluorescens
CGMCC1.1802.
Embodiment 1 prepares the complex micro organism fungicide of the present invention using following steps：
First, by bacillus subtilises, Bacillus licheniformis, aspergillus niger, colloid bacillus cereus, thermophilic coccus, enterococcus faecalis, plant
Lactobacilluss, bacillus polymyxa, bacillus megaterium, 10 kinds of bacterial strains of pseudomonas fluorescens carry out screening and the culture of bacterial strain respectively
Condition optimizing, obtains purification bacterial strain；Purification bacterial strain is carried out High Density Cultivation respectively, is inoculated into activation culture in shaking flask, will live
Change cultured strain be inoculated into fermentation tank be enlarged culture；After amplification culture terminates, list that amplification culture respectively is obtained
The zymocyte liquid of strain obtains the thalline dry powder of above-mentioned 10 kinds of strains through dehydrate；Comprise the following steps that：
1st, according to the following steps bacillus subtilises thalline dry powder is prepared by bacillus subtilis strain：
（1）The screening of bacillus subtilis strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.015-0.02g/L, complementary element：DDT
0.1-0.2g/L, pH 7.0-7.2, by MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, Sodium Chloride, glucose, DDT, agar are dissolved in
In 1000mL distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basal medium
When 45 DEG C ~ 50 DEG C are cooled to, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis culture still in melt and dissolved state
Base mixes, and is then returned in sterilizing flat board, culture medium thickness at least high 5mm on flat board, and the final pH of culture medium is in 7.2-7.5
（When temperature is 25 DEG C）, the solid medium flat board containing DDT is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus subtilises that ampoul tube is preserved make bacterium solution, coat the training of the solid containing DDT
On foster base flat board, shading culture 2-3 days under the conditions of being placed in 35 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing DDT, be placed in 35 DEG C -38 DEG C, 180-200r/
Under min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of DDT concentration, 37-39 DEG C
Lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of DDT；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 2-3%
Inoculum concentration, be transferred in the fluid medium containing DDT, 35 DEG C -37 DEG C, 220-250r/min shaken cultivation 1-3 days, finally
With degradation rate of the high effective liquid chromatography for measuring bacterial strain to DDT；
7）Screening is obtained to DDT degradation rate highest bacillus subtilises, is preserved using test tube slant as purification bacterial strain, is placed in
4 DEG C of storages of refrigerator；
（2）Bacillus subtilis strain is activated：By step 7）Bacillus subtilises that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Being inoculated in activation culture shaking flask, with shaking flask rotating speed as 160-200r/min, 26h-32h is cultivated at a temperature of 35-37 DEG C, obtain
Liquid fermentation seed；The activation culture based formulas of above-mentioned activation culture are：Peptone 9.25-11.25g/L, Carnis Bovis seu Bubali cream 1.25-
1.3g/L、NaCl32.5-5.5g/L, glucose 22-25g/L, nutritional solution pH7.5-7.8；
（3）Bacillus subtilises amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 2-5%；The culture medium prescription of amplification culture is：Semen Maydis powder 18-20g/L,
Semen Glycines powder 20-22g/L, fish flour 5-6g/L, K2HPO41.2-1.5g/L、MgSO4 .7H2O 0.15-0.2g/L, Calcium Carbonate 6.5-
8.0g/L, ammonium sulfate 1.5-1.65g/L；In fermentation tank, temperature control is at 37-40 DEG C, and ferment pH7-8, and mixing speed is 200-
250r/min, fermentation time 28-32h；When the bacillus subtilis bacterial content in fermentation tank reaches 1010During individual/ml, will ferment
Bacillus subtilises bacterium solution drying dehydration be prepared into bacillus subtilises thalline dry powder.
2nd, according to the following steps by Bacillus licheniformis（Bacillus licheniformis）Bacterial strain prepares lichens spore bar
Bacterium thalline dry powder：
（1）The screening of lichem bacillus strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：MnSO40.5-1.0g/L, peptone 8-10g/L, cattle
Meat extract 2-3g/L, NaCl 3-5g/L, glucose 1.5-2.5g/L, agar 15-20g/L, complementary element：Bayer 71628 0.15-
0.25g/L, pH7.0-7.2, by MnSO4, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, Bayer 71628, agar be dissolved in 1000mL steaming
In distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled to 45
DEG C ~ 50 DEG C when, add and be dissolved in the complementary element of 2mL sterile purified water, mix with the basal medium still in melt and dissolved state, incline
Note in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium is in 7.0-7.2（When temperature is 25 DEG C）, must contain
There is the solid medium flat board of Bayer 71628, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Bacillus licheniformis that ampoul tube is preserved make bacterium solution, coat consolidating containing Bayer 71628
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 30 DEG C -32 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing Bayer 71628, be placed in 30 DEG C -32 DEG C, 160-
Under 180r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of methylamine phosphorus concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate is as capable of the bacterial strain of degrading methamidophos；
6）By step 5）It is capable of the bacterial strain of degrading methamidophos, is transferred on activation medium respectively and is activated；Then will activation
Bacterium solution press the inoculum concentration of 3%-5%, be transferred in the fluid medium containing Bayer 71628,30 DEG C -32 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to Bayer 71628；
7）Screening is obtained to methamidophos degradation rate highest lichem bacillus strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus licheniformis strain is activated：By step 7）Bacillus licheniformis that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Slant strains are inoculated in activation culture shaking flask, with shaking flask rotating speed 160r/min, cultivate 16h-20h, obtain at a temperature of 28-30 DEG C
Obtain liquid fermentation seed；The activation culture based formulas of the activation culture are：Peptone 9.25-10.0g/L, Carnis Bovis seu Bubali cream 4.5-
5g/L, NaCl 4.8-5.0g/L, glucose 1.5g-2.0/L, nutritional solution pH6.5-7.2；
（3）Bacillus licheniformis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 3-5%；Amplification culture based formulas are：Semen Maydis powder 15-18g/L, Semen Glycines powder
22-25g/L、K2HPO40.08-0.20g/L、MgSO4 .7H2O 0.005-0.01g/L, yeast powder 0.1-0.15g/L；Fermentation temperature
25-40 DEG C, ferment pH value 6-8, and it is 8-10%, fermentation time 18-28h that mixing speed is 200-300r/min, air mass flow ratio；
When the Bacillus licheniformis content in fermentation tank reaches 1010During individual/ml, by the Bacillus licheniformis liquid drying for fermenting
Dehydration is prepared into Bacillus licheniformis soma powder.
3rd, according to the following steps aspergillus strain dry powder is prepared by Aspergillus niger strain：
（1）The screening of Aspergillus niger strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 5-10g/L, glucose 20-25g/L, sulfur
Sour magnesium 0.5-1.0g/L, yeast extract powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1.0-2.0g/L, agar 15-20g/L, mend
Fill composition：Glyphosate 0.15-0.25g/L, N,N'-dimethyl-.gamma..gamma.'-dipyridylium 0.2-0.5g/L, oxytetracycline 0.1-0.2g/L, by peptone, glucose,
Magnesium sulfate, yeast extract powder, dipotassium hydrogen phosphate, agar are dissolved in 1000mL distilled water, are subsequently adding 10mL glycerol, heating
Boil molten, and in 121 DEG C of high pressure steam sterilization 15min, basal medium is obtained, when basal medium is cooled to 45 DEG C ~ 50 DEG C, plus
Enter to be dissolved in the complementary element glyphosate of 2mL sterile purified water, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, oxytetracycline, with the basis culture still in melt and dissolved state
Base mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 6.5-6.8（Temperature is 25
DEG C when）, the solid medium flat board containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Aspergillus niger that ampoul tube is preserved makes bacterium solution, coats containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
On solid medium flat board, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated each bacterium colony, in the flat lining out of the solid medium containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, enters one
Step is separated and purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing in glyphosate, the fluid medium of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be placed in 30 DEG C -32 DEG C
Under 120-150r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, draws on the different inorganic salt solid medium flat board of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium concentration
Line, 30-32 DEG C of lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate as being capable of degradation of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Bacterial strain；
6）By step 5）Obtain can degradation of glyphosate, the bacterial strain of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be transferred on activation medium respectively and activated；
Then the bacterium solution of activation is pressed the inoculum concentration of 1-3%, is transferred to glyphosate, in N,N'-dimethyl-.gamma..gamma.'-dipyridylium fluid medium, 25 DEG C -28 DEG C,
160-180r/min vibration light culture 1-3 days, finally with high effective liquid chromatography for measuring bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Degradation rate；
7）Screening obtains the degradation rate highest Aspergillus niger bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, adopts as purification bacterial strain
Preserved with test tube slant, be placed in 4 DEG C of storages of refrigerator；
（2）Aspergillus niger strain is activated：By step 7）The aspergillus nigers for being placed in 4 DEG C of storages of refrigerator are inoculated into work in activation culture shaking flask
Change, activation culture based formulas are peptone 5.0-5.5g/L, glucose 10.0-15g/L, magnesium sulfate 0.5-0.8g/L, yeast
Powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1-3g/L, nutritional solution pH7.0-7.2 is leached, shaking flask rotating speed is 100-160r/min,
25h-28h is cultivated at a temperature of 28-35 DEG C, obtain liquid fermentation seed；
（3）Aspergillus niger amplification culture, dehydrate：By step（2）Obtain the fermentation that liquid fermentation seed is inoculated into amplification culture
Ferment in tank, be enlarged culture, inoculum concentration 8-10%；Amplification culture based formulas Rhizoma Solani tuber osi 35-40g/L, glucose 20-50g/
L, dipotassium hydrogen phosphate 5-8g/L, magnesium sulfate 1-1.5g/L, pH value are 7.0；Fermentation temperature is 35 DEG C, mixing speed 100-180r/
Min, fermentation time 25-28h, when the aspergillus niger content in fermentation tank reaches 1010During individual/ml, by the Aspergillus niger for fermenting
The dehydration of liquid drying is prepared into aspergillus strain dry powder.
4th, according to the following steps by colloid bacillus cereus（Bacillus mucilaginosus Krassilnikov）Bacterial strain system
Standby colloid bacillus cereus thalline dry powder：
（1）The screening of colloid bacillus cereus bacterial strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 2-3g/L, ammonium sulfate 0.15-0.25g/L,
Yeast extract 0.1-0.g/L, KCl0.01-0.02g/L, MgSO4 .7H2O 0.01-0.02g/L、NaH2PO40.01-0.02g/L、
CaCO30.1-0.2g/L, silica 1-2g/L, agar 10-15g/L, complementary element：Cypermethrin 0.1-0.3g/L,
CuSO4 .5H2O 0.02-0.025g/L, initial pH 7.0-7.2；By glucose, ammonium sulfate, yeast extract, KCl, MgSO4 .7H2O、
NaH2PO4、CaCO3, silicon dioxide, CuSO4 .5H2O, agar are dissolved in 1000mL distilled water, heating boil molten, and at 121 DEG C
High pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL sterilizing
The complementary element cypermethrin of distilled water, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, training
Foster base thickness at least high 5mm, the final pH of culture medium is in 7.0-7.5（When temperature is 25 DEG C）, must have cypermethrin and copper sulfate
Solid medium flat board, be placed in dark place, in 3 DEG C ~ 6 DEG C preserve；
2）Inoculation and purification：The common colloid bacillus cereus that ampoul tube is preserved make bacterium solution, coat containing cypermethrin and
On the solid medium flat board of copper sulfate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of cypermethrin concentration, 32-
35 DEG C of lucifuge cultures 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of cypermethrin；
6）By step 5）Obtain degrading the bacterial strain of cypermethrin, is transferred on activation medium respectively and is activated；Then
The bacterium solution of activation is pressed the inoculum concentration of 2-5%, has been transferred in cypermethrin fluid medium, 37 DEG C -40 DEG C, 200-220r/
Min shaken cultivation 1-3 days, finally uses degradation rate of the high effective liquid chromatography for measuring bacterial strain to cypermethrin；
7）Screening obtains the degradation rate highest colloid bacillus cereus bacterial strain to cypermethrin, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Colloid bacillus cereus actication of culture：By step 7）Colloid bacillus cereus inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Starch 15-20g/L, dipotassium hydrogen phosphate g/L, ammonium sulfate 2-3g/
L, yeast extract 3.5-5.5g/L, sucrose 8.5-10g/L, magnesium sulfate 4.5-5g/L, Calcium Carbonate 2.5-3g/L, silicon dioxide 6.5-
8g/L, ferric chloride 0.05-lg/L, pH7.0-7.2, shaking flask rotating speed is to cultivate at a temperature of 30-35 DEG C
18-20h, obtains liquid fermentation seed；
（3）Colloid bacillus cereus amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 4-6%；The composition of fermentation medium is sucrose 2.5-6.5g/L, phosphorus
Sour hydrogen dipotassium 3-5.5g/L, yeast extract 3-6g/L, ammonium sulfate 2.5-5g/L, soy peptone 5-10g/L, magnesium sulfate 3-6/g/
L、pH7.2-7.5；Amplification culture condition：32 DEG C of fermentation temperature, pH7.2, mixing speed 180r/min-220r/min, during fermentation
Between 24h；When the colloid bacillus cereus content of fermentation tank reaches 1010During individual/ml, by the colloid bacillus cereus bacterium solution warp for fermenting
Drying and dehydrating is prepared into colloid bacillus cereus thalline dry powder.
5th, according to the following steps streptococcus thermophiluss thalline dry powder is prepared by strains of streptococcus thermophilus：
（1）The screening of strains of streptococcus thermophilus is carried out according to a conventional method with training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 2-3g/L, peptone 2-3g/
L, yeast extract 1-2g/L, Fructus Lycopersici esculenti juice 15-20g/L, glucose 2-4g/L, tween 0.5ml-1.0ml/L, Calcium Carbonate 1.0-2.0g/
L, agar 2-5g/L, pH6.5-7.0, take suitable quantity of water in pot according to ratio, heating, by Carnis Bovis seu Bubali cream, peptone, yeast extract, kind
Tomato juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour in the conical flask for added agar and add after medicine boiling in pot
Plug, wrapping, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator culture 24h, asepsis growth person
Can use；The culture medium using when can also make fluid medium without agar, for the shaken cultivation of mushroom；
The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common streptococcus thermophiluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, culture obtains bacterial strain；
6）By step 5）Each bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 5%-
8% inoculum concentration, is transferred in fluid medium, 37 DEG C, quiescent culture 1-2 days, determines the biological active matter produced by each bacterial strain
Matter content；
7）Screening obtains producing bioactive substance content highest strains of streptococcus thermophilus, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Streptococcus thermophilus strain is activated：By step 7）Streptococcus thermophiluss obtaining, being placed in 4 DEG C of storages of refrigerator are inoculated into work
Change activation culture in culture shaking flask, activation culture based formulas are：Tryptone 1-3.0g/L, fish peptone 2-3g/L, Carnis Bovis seu Bubali cream
2-3g/L, yeast extract 0.5-1.5g/L, magnesium sulfate 0.3-0. 5g/L, potassium dihydrogen phosphate 0.5-1.0g/L, Lactose 1-1.5g/L,
Nutritional solution pH 6.5-6.8, cultivates 24h-28h at a temperature of 35-40 DEG C, and mixing speed is 80-100r/min, obtains liquid and sends out
Ferment seed；
（3）Streptococcus thermophiluss amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 2-5%；Amplification culture based formulas：Glucose 1-3g/L, soy peptone 4-
6g/L, dipotassium hydrogen phosphate 2-3g/L, corn juice 40-50g/L, sodium acetate 1-2g/L, skimmed milk 10-15g/L, pH are 6.5-6.8；
38-45 DEG C of fermentation temperature, mixing speed is 100-120r/min, fermentation time 48-72h；When the streptococcus thermophiluss in fermentation tank
Content reaches 109During individual/ml, the streptococcus thermophiluss bacterium solution drying for fermenting dehydration is prepared into streptococcus thermophiluss thalline and is done
Powder.
6th, by E. Faecium strains, comprising the following steps that for enterococcus faecalis thalline dry powder is prepared according to the following steps：
（1）The screening of E. Faecium strains and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 10-15g/L, maltose 20-25g/L, ferment
Female leaching powder 10-2g/L, sodium glycerophosphate 5-6g/L, Sodium Chloride 1-2g/L, Lactose 0.015-0.020g/L, agar 15-20, pH
Value 8.0-8.5, is separately separately added into complementary element in order in the medium：Erythromycin 0.05-0.15g/L, ciprofloxacin
0.01-0.02g/L, Sulfamethoxazole Compound 0.25-0.35g/L；By peptone, maltose, yeast extract, sodium glycerophosphate,
Sodium Chloride, Lactose, agar are dissolved in 1000mL distilled water, are subsequently adding 10mL glycerol, and heating is boiled molten and high at 121 DEG C
Pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL sterilizing steaming
The complementary element erythromycin 0.05-0.15g/L of distilled water, ciprofloxacin 0.01-0.02g/L, Sulfamethoxazole Compound 0.25-
0.35g/L, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, and culture medium thickness are at least high
5mm, the final pH of culture medium is in 7.5-8.0（When temperature is 25 DEG C）, containing erythromycin, ciprofloxacin, compound sulfonamide first dislike
Azoles solid medium flat board, is placed in dark place, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common enterococcus faecalis that ampoul tube is preserved make bacterium solution, coat husky containing erythromycin, ring third
On star, Sulfamethoxazole Compound solid medium flat board, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium
Fall；
3）By step 2）Isolated bacterium colony, containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound solid medium
Flat lining out, is separated and purification further；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound liquid culture
In base, 33 DEG C -35 DEG C are placed in, under 160-180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, solid in the different inorganic salt of erythromycin, ciprofloxacin, Sulfamethoxazole Compound concentration
Rule on body culture medium flat plate, 35 DEG C -38 DEG C lucifuge cultures 2-5 days；The bacterial strain that can grow on culture medium flat plate is energy
Enough degraded erythromycin, ciprofloxacin, the bacterial strain of Sulfamethoxazole Compound；
6）By step 5）Obtain degrading the bacterial strain of erythromycin, ciprofloxacin, Sulfamethoxazole Compound, is transferred to activation respectively
Activated in culture medium, then the bacterium solution of activation pressed the inoculum concentration of 3%-5%, be transferred to pH8.5-9.0 containing erythromycin,
In ciprofloxacin, Sulfamethoxazole Compound element fluid medium, at 38 DEG C -40 DEG C, 180-250r/min vibrates light culture 1-3
My god, finally with each bacterial strain of high effective liquid chromatography for measuring to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate；
7）Screening obtain to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate highest E. Faecium strains, as
Purification bacterial strain is preserved using test tube slant, is placed in 4 DEG C of storages of refrigerator；
（2）Enterococcus faecalis actication of culture：By step 7）The enterococcus faecalis for being placed in 4 DEG C of storages of refrigerator for obtaining are inoculated into activation culture
Activate in shaking flask, activation culture based formulas are：Glucose 4-5g/L, yeast powder 0.15-0.3g/L, peptone 0.5-1g/L, no
Water calcium chloride 0.002-0.006g/L, bitter salt 0.02-0.05g/L, dipotassium hydrogen phosphate 0.1-0.15g/L, seven hydrations
Manganese sulfate 0.002-0.005g/L, Calcium Carbonate 0.5-1g/L, nutritional solution pH7.5-8.0, shaking flask rotating speed is 180-200r/min,
30h-35h is cultivated at a temperature of 30-35 DEG C, obtain liquid fermentation seed；
（3）Enterococcus faecalis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into sending out for amplification culture
Ferment in fermentation tank, be enlarged culture, inoculum concentration 8-10%；Culture medium prescription is tryptone 10-15g/L, yeast powder 10-
15g/L, glucose 10-15g/L, anhydrous calcium chloride 0.04-0.06g/L, bitter salt 0.019-0.025g/L, phosphoric acid hydrogen
Dipotassium 0.04-0.06g/L, 0.04-0.06g/L of potassium dihydrogen phosphate, dibastic sodium phosphate 0.4-0.6g/L, Sodium Chloride 0.08-0.1g/
L, murphy juice 50-80g/L, Calcium Carbonate 1-3g/L, sodium acetate 10-15g/L, ammonium carbonate 2-3g/L, ammonium acetate 1-2g/L, half Guang ammonia
Acid hydrochloride 0.5-1g/L, pH value is 8.5-9；35-37 DEG C of fermentation temperature, mixing speed 180-220r/min, fermentation time 20-
32h, when the enterococcus faecalis content of fermentation tank reaches 1010During individual/ml, by the enterococcus faecalis bacterium solution drying for fermenting dehydration system
Standby become enterococcus faecalis thalline dry powder.
7th, according to the following steps Lactobacillus plantarum thalline dry powder is prepared by lactobacillus plantarum strain：
（1）The screening of lactobacillus plantarum strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 1-2g/L, peptone 1-2g/
L, yeast extract 1-2g/L, 20-30g/L of Fructus Lycopersici esculenti juice, glucose 1-2g/L, tween 0.05ml-0.10ml/L, Calcium Carbonate 1.5-
2.0g/L, agar 2-5g/L, pH6.2-6.5, according to ratio according to the required suitable quantity of water that measures in pot, heating, weigh Carnis Bovis seu Bubali cream,
Peptone, yeast extract, Fructus Lycopersici esculenti juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour into and add after medicine boiling in pot
Jump a queue in the conical flask of agar, wrap up, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator trainings
Foster 24h, asepsis growth person can use；Culture medium using when can also make fluid medium without agar, for bacterium
The shaken cultivation of class；The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common plant lactobacilluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）The each bacterial strain for obtaining, is transferred on activation medium respectively and is activated, and then presses the bacterium solution of activation
The inoculum concentration of 3%-5%, is transferred in fluid medium, 37 DEG C, and quiescent culture 1-2 days finally determines the life produced by each bacterial strain
Active substances content；
7）Screening obtains producing bioactive substance content highest lactobacillus plantarum strain, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Lactobacillus plantarum actication of culture：By step 7）Lactobacillus plantarum obtaining, being placed in 4 DEG C of storages of refrigerator is inoculated into work
Change and activate in culture shaking flask, activation culture based formulas are：Peptone 10-12g/L, Carnis Bovis seu Bubali cream 10-15g/L, yeast extract 5-6.5g/
L、KH2PO42-3.5g/L, trisodium citrate 2-2.5g/L, sodium acetate 2-2.5g/L, glucose 20-28g/L, MgSO4 .7H2O
0.58-0.60g/L、MnSO4 .4H2O 0.25-0.3g/L, nutritional solution pH 6.2-6.5, cultivate 24h- at a temperature of 35-40 DEG C
28h, quiescent culture, obtain liquid fermentation seed；
（3）Lactobacillus plantarum amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 5-8%；Culture medium prescription is：Sucrose 25-30g/L, yeast extract 15-20g/
L、KH2PO48-10g/L, trisodium citrate 5-6g/L, sodium acetate 3-5g/L, MgSO4 .7H2O 0.8-1.2g/L、MnSO4 .4H2O
0.5-0.8g/L, pH 6.2-6.5；35-37 DEG C of fermentation temperature, fermentation time 24-28h, quiescent culture；Fermentation ends, will fermentation
Good Lactobacillus plantarum bacterium solution drying dehydration is prepared into Lactobacillus plantarum thalline dry powder.
8th, according to the following steps bacillus polymyxa thalline dry powder is prepared by bacillus polymyxa bacterial strain：
（1）The screening of bacillus polymyxa bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-15g/L, yeast extract 5-8g/
L、NaCl3-5g/L、KH2PO40.1-0.2g/L、MgSO4 .7H2O 0.05-0.1g/L、MnSO4 .7H2O 0.25-0.5g/L, fine jade
Fat 15-20g/L, pH7.5-8.0；By MnSO4 .7H2O, peptone, KH2PO4、NaCl、MgSO4 .7H2O, agar are dissolved in 1000mL
In distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled to
When 45 DEG C ~ 50 DEG C, it is poured in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium is in 7.5-8.0（Temperature
During for 25 DEG C）, solid medium flat board is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus polymyxa that ampoul tube is preserved makes bacterium solution, coats solid medium flat board
On, shading culture 2-3 days under the conditions of being placed in 37 DEG C -40 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single dominant bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be placed in 38 DEG C -40 DEG C, 160-180r/min vibrate bar
Under part, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 38-40 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 2-5%
Inoculum concentration, be transferred in fluid medium, 37 DEG C -40 DEG C, 180-200r/min shaken cultivation 1-3 days, finally determine each bacterium
The various bioactivators contents such as such as antibiotic, antagonist protein, phytohormone, enzyme, flocculant produced by strain；
7）Screening obtains producing bioactive substance content highest bacillus polymyxa bacterial strain, adopts test tube as purification bacterial strain
Inclined-plane is preserved, and is placed in 4 DEG C of storages of refrigerator；
（2）Bacillus polymyxa actication of culture：By step 7）Screening obtains, is placed in the bacillus polymyxa of 4 DEG C of storages of refrigerator and connects
Plant and activate in activation culture shaking flask, activation culture based formulas are：Peptone 8-10g/L, Carnis Bovis seu Bubali cream 3-5g/L, Sodium Chloride 3-
5g/L, glucose 10-12g/L, KH2PO40.2-0.3g/L、MgSO4 .7H2O 0.2-0.35g/L、NaCl 0.2-0.3g/L、
CaSO4 .2H2O 0.2-0.25g/L、CaCO35.0-6.5g/L, nutritional solution pH 7.0-8.0, shaking flask rotating speed 150-170r/min,
25h-30h is cultivated at a temperature of 37-40 DEG C, obtain liquid fermentation seed；
（3）Bacillus polymyxa amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 10-12%；Amplification culture based formulas are：Semen Maydis powder 20-25g/L, Semen Glycines powder
8-12g/L、K2HPO40.05-0.15g/L, Sodium Chloride 1.0-1.5g/L, Na2HP040.15-0.25g/L；Fermentation temperature 28-
35 DEG C, ferment pH7-8, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h；When the bacillus polymyxa of fermentation tank contains
Amount reaches 1010During individual/ml, the bacillus polymyxa bacterium solution drying for fermenting dehydration is prepared into bacillus polymyxa thalline
Dry powder.
9th, according to the following steps bacillus megaterium thalline dry powder is prepared by bacillus megaterium bacterial strain：
（1）The screening of bacillus megaterium bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.025-0.035g/L, complementary element：To sulfur
Phosphorus 0.15-0.20g/L, pH7.5-8.0；By MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, parathion, agar are molten
Solution is in 1000mL distilled water, and heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basic training
When foster base is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis still in melt and dissolved state
Culture medium mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 7.5-8.0（Temperature
During for 25 DEG C）, the solid medium flat board containing parathion is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus megaterium that ampoul tube is preserved makes bacterium solution, coats consolidating containing parathion
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of parathion concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of parathion；
6）By step 5）Obtain degrading the bacterial strain of parathion, is transferred on activation medium respectively and is activated, then will
The bacterium solution of activation presses the inoculum concentration of 2-5%, has been transferred in parathion fluid medium, and 37 DEG C -40 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to parathion；
7）Screening obtains the degradation rate highest bacillus megaterium bacterial strain of parathion, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus megaterium actication of culture：By step 7）Bacillus megaterium inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Peptone 10-15g/L, Carnis Bovis seu Bubali cream 3-5g/L, NaCl6.5-
8.5g/L, glucose 2.5-3g/L, nutritional solution pH7.0-7.2, shaking flask rotating speed is to cultivate 190-210r/min, at 37-40 DEG C
22-28h, obtains liquid fermentation seed；
（3）Bacillus megaterium amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5-9%；The culture medium prescription of amplification culture is：Semen Maydis powder 15-18g/L,
Semen Glycines powder 25-30g/L, fish flour 5-6g/L, K2HPO40.05-0.15g/L, calcium chloride 5.5-6.5.0g/L, ammonium sulfate 2.0-2.5g/
L；36-40 DEG C of fermentation temperature, fermentation pH value issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss；
When the bacillus megaterium content in fermentation tank reaches 1010During individual/ml, by the bacillus megaterium bacterium solution drying for fermenting
Dehydration is prepared into bacillus megaterium thalline dry powder.
10th, according to the following steps pseudomonas fluorescens thalline dry powder is prepared by pseudomonas fluorescens strain：
（1）The screening of pseudomonas fluorescens strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-12g/L, K2SO410-12g/L、
MgCl21.4-1.6g/L、Fe2(SO4)30.5-1g/L, glycerol 10mL, 15 ~ 20g/L of agar, distilled water 1L, complementary element：
Penicillin 0.05-0.10g/L；By tryptone, K2SO4、MgCl2、Fe2(SO4)3, agar be dissolved in 1000mL distilled water,
It is subsequently adding 10mL glycerol, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, basal medium is obtained, treats basic culture
When base is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the basis training still in melt and dissolved state
Foster base mixing, is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is in 7.0-7.2（Temperature is
When 25 DEG C）, the solid medium containing penicillin is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common fluorescent pseudomonass that ampoul tube is preserved make bacterium solution, coat consolidating containing penicillin
On body culture medium flat plate, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing penicillin, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing penicillin, be placed in 25 DEG C -28 DEG C, 150-
Under 180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of penicillin concn, 25-28
DEG C lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of penicillin；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 1%-3%
Inoculum concentration, be transferred to containing in penicillin fluid medium, at 25 DEG C -28 DEG C, 160-180r/min vibrates light culture 1-3
My god, finally use degradation rate of the high effective liquid chromatography for measuring bacterial strain to penicillin；
7）Screening is obtained to penicillin degradation rate highest pseudomonas fluorescens strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Pseudomonas fluorescens actication of culture：By step 7）The pseudomonas fluorescens inclined-plane bacterium for being placed in 4 DEG C of storages of refrigerator for obtaining
Plant to be inoculated in activation culture shaking flask and activate, activation culture based formulas are：Carnis Bovis seu Bubali cream 2.55-3.0g/L, peptone 1.65-
2.2g/L, Sodium Chloride 0.18g-0.2g/L, yeast extract 5.25-6.0g/L, Fe2(SO4)31-1.5g/L, culture fluid pH7.0-
7.5, shaking flask rotating speed is to cultivate 18-22h at a temperature of 28.5-31 DEG C, obtains liquid fermentation seed；
（3）Pseudomonas fluorescens amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5.5 ~ 8.5%；The culture medium prescription of the amplification culture is：Maltose
20-35g/L, Semen Maydis powder 12-15g/L, yeast extract 4.5-4.62g/L, ammonium sulfate 5.2-5.8g/L, K2HPO40.55-0.85g/
L、MgSO4 .7H2O 0.035-0.055g/L、FeSO4 .7H2O 1.095-1.20g/L、CaCl20.075-1.0g/L、NaCl
0.15-0.25g/L；27 DEG C ~ 29 DEG C of fermentation temperature, fermentation tank 140 ~ 180r/min of mixing speed, ventilation 8 ~ 12%, during fermentation
Between 24 ~ 48h；When the Pseudomonas bacterial content in fermentation tank reaches 1010During individual/ml, by the pseudomonas fluorescens for fermenting
The dehydration of bacterium solution drying is prepared into pseudomonas fluorescens thalline dry powder.
2nd, the thalline dry powder of 10 kinds of purification bacterial strains for preparing step one uniformly mixes, and obtains many thalline compound bacteria
Powder；In many thalline composite bacterium powder, in each strain thalline dry powder, live body bacterium number accounts for the weight percent of composite bacterium powder total viable count
Than as follows：Bacillus subtilises 10-15%, aspergillus niger 10-15%, colloid bacillus cereus 10-15%, enterococcus faecalis 10-15%, lichens
Bacillus cereuss 8-12%, bacillus megaterium 8-12%, pseudomonas fluorescens 8-12%, Lactobacillus plantarum 5-8%, bacillus polymyxa
5-8%, streptococcus thermophiluss 6-8%.In many thalline composite bacterium powder, effective active bacterial content is hundred million cfu/g of 200-500, water
Divide content 5%；Filtrated air packaging is passed through in many thalline composite bacterium powder, obtains final product complex micro organism fungicide.
Embodiment 2 is accounted for the percentage by weight of composite bacterium powder total viable count by live body bacterium number in each strain thalline dry powder：Hay
Bacillus cereuss 15%, aspergillus niger 15%, colloid bacillus cereus 15%, enterococcus faecalis 15%, Bacillus licheniformis 8%, bacillus megaterium
8%th, pseudomonas fluorescens 8%, Lactobacillus plantarum 5%, bacillus polymyxa 5%, streptococcus thermophiluss 6%.Made using embodiment step one
The thalline dry powder blend of the standby 10 kinds of purification bacterial strains for obtaining is uniform, much thalline composite bacterium powder, is passed through filtrated air, and packaging is obtained
Complex micro organism fungicide.
Embodiment 3 is accounted for the percentage by weight of composite bacterium powder total viable count by live body bacterium number in each strain thalline dry powder：:Hay
Bacillus cereuss 10%, aspergillus niger 10%, colloid bacillus cereus 10%, enterococcus faecalis 10%, Bacillus licheniformis 12%, huge spore bar
Bacterium 12%, pseudomonas fluorescens 12%, Lactobacillus plantarum 8%, bacillus polymyxa 8%, streptococcus thermophiluss 8%.Using embodiment step
The thalline dry powder blend of the one 10 kinds of purification bacterial strains for preparing is uniform, and much thalline composite bacterium powder, is passed through filtrated air, packaging
Prepared complex micro organism fungicide.
Embodiment 4 is accounted for the percentage by weight of composite bacterium powder total viable count by live body bacterium number in each strain thalline dry powder：Hay
Bacillus cereuss 12%, aspergillus niger 12%, colloid bacillus cereus 12%, enterococcus faecalis 12%, Bacillus licheniformis 10%, huge spore bar
Bacterium 10%, pseudomonas fluorescens 10%, Lactobacillus plantarum 7%, bacillus polymyxa 7%, streptococcus thermophiluss 8%.Using embodiment step
The thalline dry powder blend of the one 10 kinds of purification bacterial strains for preparing is uniform, and much thalline composite bacterium powder, is passed through filtrated air, packaging
Prepared complex micro organism fungicide.
The complex micro organism fungicide of the present invention is a kind of multi-functional product, is now applied to be illustrated below.
The complex micro organism fungicide of 5 present invention of embodiment can be applicable to carry out feces of livestock and poultry at the innoxious place of compost fermentation
Reason, solves the problems, such as antibiotic with drug residue.This kind of application mode is not limited to the innoxious place to aquaculture feces of livestock and poultry
Reason, is also applied for the innoxious of each type organic of agriculture waste, industrial processes Organic Ingredientss, organic waste, organic sludge etc.
Process.Concrete application example is as follows.
Livestock and poultry cultivation enterprise zone is cultivated in Fujian, Guangdong, Henan, Chongqing, Sichuan, Shandong, according to cultivation scale in poultry
1000-50000 ten thousand（Ten thousand plumage of fowl 5000-100000）Plant extract 100 animal dung samples（Wherein 65, individual pig manure sample,
35 chicken manure samples）, application conventional chemical analysis method determines nutrient therein, antibiotic, antibiotics and content of beary metal, same
Shi Yingyong Solid-Phase Extraction-high performance liquid chromatography tandem mass spectrum method has carried out special measure to residual antibiotic therein.
As a result show：Containing the nutrient such as abundant organic matter and total nitrogen, total phosphorus, total potassium in plant's animal dung；Cr、Ni、
The contents of heavy metal elements such as Cd, Hg, Pb are relatively low, but Cu, Zn content is universal higher, and As, Cu and Zn content is respectively 0.654-
81.92 mg/kg, 18.50-631.98 mg/kg and 85.06-289.92 mg/kg.With China agricultural sludge pollutant catabolic gene
National standard (GB 4284-1984) is compared, and in animal dung, the exceeding standard rate of As, Cu and Zn is extremely serious.In 65 pig manure samples, its
In：In 35 samples, oxytetracycline, tetracycline, the average content of chlortetracycline are respectively 11.13,7.31,5.52 mg/kg；20 samples
In this, oxytetracycline, tetracycline, the average content of chlortetracycline are respectively 19.12,10.35,70.55 mg/kg；Soil in 10 samples
Mycin, tetracycline, the average content of chlortetracycline are respectively 36.03,49.34,191.12 mg/kg.In 35 chicken manure samples, soil is mould
Element, tetracycline, average content respectively 6.91,8.52,9.47 mg/kg of chlortetracycline, quinolones and sulfadimidine
Content is respectively 20.93-66.62 mg/kg.In feces of livestock and poultry, other such as the recall rate such as penicillin, streptomycin be 100%,
65.3%.Therefore, in plant's animal dung harmful components to there is potentially hazardous property after using in farmland, using front reply poultry
Excrement carries out harmless treatment.
Complex micro organism fungicide of the present invention can intensive decomposition protein, cellulose, hemicellulose, lignin etc., and will
Thermophilic, thermoduric bacteria, funguses, yeast strain and correlation down enzyme are composited, its living bacteria count content height, degradation capability
By force, while intensification, deodorization can be reached, and eliminate pest and disease damage, weed seed and improves the effect of nutrient.Under appropriate conditions,
Rapidly the carbon in windrow, nitrogen, phosphorus, potassium, sulfur etc. can be decomposed mineralising, degraded forms simple organic, so as to be further broken into
The absorbable nutritional labeling of crop.
The complex micro organism fungicide of the present invention is applied to the innoxious of feces or sludge or organic waste or industrial organic waste
Process, can be performed as follows：
（1）Prepare treating material according to following quality proportioning：Major ingredient is that feces of livestock and poultry or sludge or organic waste or industry have
Machine waste material 70%-80%, adjuvant is the 20%-30% such as dry Testa oryzae, sawdust, grouts powder, straw stalk powder；
（2）According to the percentages for the treatment of material weight, complex micro organism fungicide in the inoculum concentration for the treatment of material is
0.02%-0.1%；
（3）Material mixing and control and regulation：By major ingredient, adjuvant and complex micro organism fungicide mix, moisture Control in 50%-60%,
Handss grab the agglomerating no water droplet of material, loose one's grip i.e. and dissipate for standard；According to ten thousand/carbamide to 10/10000ths or so adjusts carbon-nitrogen ratio
To 25-30:1（Adjust the nitrogen of carbon-nitrogen ratio, it is also possible to substituted with the available nitrogen source of other plant）；Stack height is built for 1.2-1.6
Within rice, width is not limited with length；Heap optimal volume is built in 100-1000 cubic meter；Ordinary circumstance, when building heap, ambient temperature is
Room temperature；Winter builds heap ambient temperature more than 0 DEG C, and suitable environment temperature is can to eliminate stink in more than 15 DEG C, 2-3 days.Build heap
When heap temperature reaches 55 DEG C -68 DEG C and continues more than 48 hours afterwards, turning is once；Sustainable in 55 DEG C -68 DEG C degree high temperature
More than 10 days, the pathogenic bacteria in killing fermented product, worm's ovum, weed seed, one week or so summer became thoroughly decomposed completely, and 15 days or so winter is complete
Become thoroughly decomposed entirely；
（4）Antibiotic is degraded with drug residue：After above-mentioned material becomes thoroughly decomposed, using anaerobism and microaerophilic control mode, carry out
Heap body material latter stage of ripening；Heap body not being stirred in 7-10 days, high temperature bacterium stops growing, enters rest period；In the later stage by low temperature
Anaerobe with and anaerobe control heap temperature at 35 DEG C or so, allow the various funguses of heap body, antibacterial etc. set up small-sized microorganism
Group's system, with organic matter and other nutrient sources of heap body as common main foundation energy source, using good alternate and Co metabolism
Relation, degraded antibiotic and drug residue.After -15 after 10 days, heap body completes after-ripening, and antibiotic is substantially all with drug residue
Degraded, different antibiotic reach 99.5-100% with chemicalses degradation rate.
Using above-mentioned step, the innoxious organic substance that becomes thoroughly decomposed has the advantage that, and can be further with：
（1）The innoxious organic substance that becomes thoroughly decomposed can be used directly as nontoxic, harmless, free of contamination high quality organic fertilizer material, should
The equilibrium of material nutrient, total nutrient loss is few, humus content height, and N-P-K content increases；
（2）A large amount of function bacterium being bred in sweat and produces multiple specially good effect metabolites, so as to stimulate crop growth, carries
High crop is disease-resistant, drought resisting, cold tolerance, function antibacterial enter soil after, can fixed nitrogen, phosphorus decomposing, potassium decomposing, increase soil nutrient, change
Good Soil structure, raising chemical fertilizer utilization ratio；
（3）Material is through mineralization, and nutrient is changed into available state and quick-acting states from invalid state and slow state；Through humification, produce
A large amount of humic acid, stimulate plant growth.
The complex micro organism fungicide of the present invention of embodiment 6 in agricultural planting improved soil, carry out Biological control, reduce agriculture
Industry environmental pollution, the application note for increasing crop yield and improving crop product quality are as follows.
1st, the multifunctional composite microbe microbial inoculum of the present invention is made up of 10 kinds of antibacterials or funguses, the function of its single bacterial strain with
Key property is as follows.
（1）Pseudomonas fluorescens：Its characteristic is to have good Degradation to penicillium sp with function, with decomposition fiber
Element, protein and combine Fe3+Ability；Pseudomonas fluorescens can also produce chitinase, lysozyme, xylanase and extracellular shell
Dextranase etc. is degraded to material, can also reduce the generation of plant disease while nutrition is provided to plant；Can carry out anti-
Nitrification, decomposable asymmetric choice net fat.
（2）Bacillus licheniformis：Its characteristic is with function for producing various active enzyme, such as protease, amylase, fat
The mechanism of its Biocontrol Effect such as enzyme, pectase, glucanase, cellulase is to compete, based on antagonism and inducing plant resistance, ground
The physiological feature of clothing bacillus cereuss is rich and varied, widely distributed, easily separation and Culture, it is easy in soil and plant surface, root
Border forms superior microorganism population, so as to effectively suppress with identical nutrition and the growth of space site pathogen and breeding.
The antagonistic substance that Bacillus licheniformis produce mainly has antibiotic, bacteriocin, cell wall degradation enzyme and other antibacterial proteins, right
Antibiotic has certain degradation function with residual pesticide.Meanwhile, Bacillus licheniformis have stronger protease, amylase and fat
Fat enzymatic activity, can not only suppress or murder pathogen, and the larger molecular organicses in soil of can degrading, and activating soil is supported
Point, contribute to the absorption of plant nutrient, improve crop yield, improving quality.It is discharged in agricultural environment, the micro- life of soil can be improved
State environment, makes that the dominant population in soil is stable to keep on top.After Bacillus licheniformis are manured into soil, rapid breeding
100 times become dominant bacteria, control nutrition and other resources of rhizosphere, cause source of disease bacterium to lose vivosphere to a great extent
And condition；Improve plant related organization cell wall thickening, fibrosiss, degree of lignification, and it is double that cutin is formed outside epidermal area
Silicon layer, forms the barrier for preventing pathogenic bacteria invasion and attack together.While the supply of nitrogen can be improved, and phosphorus decomposing, potassium decomposing, can effectively improve in soil
Phosphorus and the absorbance of potassium.
（3）Bacillus subtilises：Its characteristic is with function for producing multiple antibiotics and raw albumen enzyme, α-amylase, fibre
Ten several enzymes such as the plain enzyme of dimension, 1,4 beta-glucanase, phytase, pectase, xylanase, with broad spectrum antibiotic activity and extremely strong anti-
Inverse ability.The sporinite of bacillus subtilises can successfully be converted into trophosome and occupy advantage long-term surviving and continue breeding
Play a role.Crop disease-resistant, cold-resistant, drought-resistant ability can be improved；Increase soil nutrient, improved soil structure, improve chemical fertilizer utilization
Rate；Promote the organic matter decomposition in soil to become humus, stimulate plant growth.Energy fixed nitrogen, phosphorus decomposing, potassium decomposing, improve fertilizer utilization
Rate.Can improved soil structure, promote the organic matter decomposition in soil to become humic acid, promote plant growth, maturation, reduces cost,
Increase yield, improve income.Similar cell mitogen, the material of plant growth activating element is produced, and decomposes heavy metal, pesticide
Residual.
（4）Its characteristic of bacillus megaterium with function for main by colonizing, secreting antibiotic substance and battalion in plant rhizosphere
Support and compete and antagonism pathogen.Huge spore is the circle or ellipse that antibacterial is formed in the cell in its Later growth
Resistance hypopuss.Spore has some special properties such as extremely strong heat resistanceheat resistant, radioprotective, anti-chemicalses and hydrostatic pressure resistant.
Spore is compared with its easy preservation of nutrition somatic cell, resurrection rate height.Bacillus megaterium bacillus megaterium has degraded soil well
Effect of middle organophosphors, has repair to the water body for polluting, with other bacillus cereuss of nature by symbiosis and Co metabolism,
The antibiotic for remaining in degraded environment and pesticide.
（5）Bacillus polymyxa：Its characteristic is with function for producing such as antibiotic, antagonist protein, phytohormone, enzyme, wadding
The various bioactivators such as solidifying agent.Multiple animals and plants pathogenetic bacterias and funguses are had stronger antagonism, send out by has a broad antifungal spectrum
Ferment character is excellent, can produce multi-component cyclic peptide antibacterial active substance, while antifungal active substance can be produced, have concurrently
Biological pesticide and the effect of bio-bacterial manure, are widely used to the fields such as agricultural, industry and mining and wastewater treatment.Glue class spore bar more
Bacterium is a kind of plant rhizosphere growth-promoting antibacterial, can be used as microbial manure, promotes plant growing, improves crop yield.Effectively can prevent
Vegetative bacteria and fungoids soil-borne disease is controlled, the polypeptide antibiotic substance that Paenibacillus polymyxa is produced is to plant pathogenic fungi
With good antagonistic activity, and Nature comparison is stable, can be used for the Biological control of plurality of plant diseases.
（6）Lactobacillus plantarum：Its characteristic makes the pH value in water stably not raise with function for substantial amounts of producing acid, and
The acidic materials of its output can be degraded heavy metal, Lactobacillus plantarum has that breeding is quick, vitality is strong, safety non-toxic the features such as.
Lactobacillus plantarum can secrete the multiple organic acid of synthesis, enzyme, biological active substanceies.
（7）Enterococcus faecalis：Its characteristic is to produce beneficial nutrient substance in growth and metabolic process with function, such as breast
Acid, aminoacid, vitamin, enzyme and antibacterial substance.Enterococcus faecalis form good alternate Co metabolism with bacillus cereuss and aspergillus niger
Effect, to penicillins, erythromycin, quinolones, nitrofurantoin, high concentration gentamycin and high concentration streptomycin and rifampicin
With good degradation capability.Enterococcus faecalis are aerobic and facultative anaerobic bacteria, during fermentation animal wastes, quickly can remove
Smelly, control harmful gass volatilization.
（8）Aspergillus niger：Its characteristic and function be aspergillus niger secretion produce amylase, saccharifying enzyme, citric acid, gluconic acid,
The function of gallic acid etc..Aspergillus niger has cracking larger molecular organicses and indissoluble inorganic matters ability, is easy to Crop to utilize,
Improve Soil structure, strengthen soil fertility, improve the effect of crop yield.Under other microorganism Co metabolism effects, to BAICAO
The herbicides such as withered, glyphosate and chemical pesticide have certain Degradation.
（9）Streptococcus thermophiluss：Characteristic and function are to suppress to pollute the harmful bacteria pathogen in environment, such as clostridium difficile,
Salmonella typhimurium, escherichia coli, staphylococcus aureus, Listeria monoeytogenes, bacillus perfringens etc..Sending out
During ferment animal wastes, can suppress or eliminate harmful pathogen quickly, shorten sweat, improve the ferment effect that becomes thoroughly decomposed.
Meanwhile, it is widely used in some important fermented dairy products of production.Streptococcus thermophiluss have some functional activities, such as produce extracellular
Polysaccharide, bacteriocin and vitamin.
（10）Colloid bacillus cereus：Its characteristic and function be decompose in silicate and aluminosilicate and other Ores containing potassium
Mineral, with Soluble phosphorus, Potassium release and fixed nitrogen function, while can produce organic acid, aminoacid, polysaccharide, swash during growth and breeding
Element etc. is conducive to plant absorption and the material for utilizing.After breeding in soil, plant growth substance and multiple enzymes is secreted, to increase
Resistance of the thing to some diseases is pretended, the growth of other pathogen can be suppressed.Potassium content in thalline ash more than 33%,
Endobacillary potassium directly can be absorbed by plant in separate out after thalline death.There is activation chesson, decompose respectively
Trace element, silico-calcium sulfur boron molybdenum zinc etc. in kind.The several physiological active substances such as gibberellins, heteroauxing are produced, makes plant growth
Stalwartness, strengthens the disease-resistant and anti-adversity ability of crop cold resistance drought resisting, improves crop yield and improves product quality.Formed in crop root
Profitable strain, the effectively generation of suppression soil-borne disease.Drop with certain Adsorption of Heavy Metals ability and with other microorganism Co metabolism
Solution pesticide.
2nd, above-mentioned 10 kinds of microorganism lists thalline forms complex micro organism fungicide after compound, according to the alternate of microorganism, altogether
Metabolism, synergism used in agricultural, with multi-Functional Fertilizers, biological pest control, repair agroecological environment work(
Effect and effect, concrete application is as follows：
（1）Above-mentioned complex micro organism fungicide is the composite flora of multiple beneficial microorganism, can develop green agriculture with improved soil
Industry, livestock-raising, environmental improvement and resource regeneration, are emerging biological environmental production products, meet economic development needs；This
Bright microbial composite bacteria is tested through experimental plot, and its result shows that the effect of increasing production of the like product than having reported becomes apparent from,
Disease and insect resistance is higher；Use no or little chemical fertilizer and pesticide can just improve yield, weeds and pest and disease damage is eliminated, promote well green
The development of color organic agriculture and the reparation of natural ecosystems；
（2）Complex micro organism fungicide provide can fixed nitrogen, phosphorus decomposing, the beneficial microbe such as potassium decomposing, the microorganism that these are lived can be
Plant rhizosphere growth, breeding, can bring the benefit of several respects：One is by these beneficial microorganism vital movements, fixes
The unavailable molecular nitrogen of conversion in the air be combined nitrogen, parsing unavailable compound state phosphorus in soil, potassium for can profit
With the phosphorus of state, potassium, and more than 10 kind of middle and trace element in soil can be parsed；Two are lived by these beneficial microorganism life
Dynamic, the phytohormone such as secretion auxin, the basic element of cell division, gibberellins, indolic acid, promote plant growth, regulate and control crop metabolism, press
Genetic code builds quality product；Three is in rhizosphere amount reproduction by beneficial microbe, produces a large amount of mucopolysaccharides, divides with plant
The mucus that secretes and mineral gels, organic colloid combine, and form soil aggregate, promote soil and store fertilizer, water holding capacity.Four are
Can promote crop growth, improved soil structure, improve crop product quality and the diseases prevention of crop, resistances against diseases are improved, so as to
Realize increasing both production and income；Four is by microorganism alternate and Co metabolism, solves antibiotic and pesticide residual contamination.
3rd, it is applied to improved soil in agricultural planting, carries out Biological control, drop using the complex micro organism fungicide of the present invention
When low agricultural environment pollution, increase crop yield and the application of raising crop product quality, using method consumption is as follows with scope：
The multifunctional microbial bacterium composite bacteria agent capable of the present invention is applied to whole proportion of crop plantings and natural environment pollution control, in crops
During site preparation, together use as base manure with organic fertilizer, per mu of crops use 1-2 kilogram；Or after diluting 100 times with carrier
Directly it is manured into soil after mixing, can be used in mixed way with other chemical fertilizer；When using as topdressing, 500- is directly diluted to
800 times of liquid carry out sprinkler irrigation fertilization, and per mu of crops consumption is 300-500 gram.
Points for attention：Can not use with antibacterial, pestsides synthesis.When using as multi-Functional Fertilizers, typically can not be independent
Apply, can chemical fertilizer, fertilizer compounding application, can just give full play to the volume increase efficiency of multifunctional microbial bacterium composite bacteria agent capable.And
Certain environmental condition and cultivation step is required, to ensure the environmental condition required for microbial growth, makes fertilizer abundant
Play a role.
The complex micro organism fungicide of 7 present invention of embodiment in degrading pesticide and processes a huge sum of money as agricultural environment renovation agent
The method that category pollution is used is as follows with application note.
The complex micro organism fungicide degrading pesticide of the present invention and the know-why of antibiotic remainss, one be using microorganism it
Between intergrowth relation, set up the ecological balance of microbiologic population, acted on using microorganism Co metabolism, supplement in agricultural environment multiple
Close microbial bacteria and environment indigenous microorganisms symbiosis, set up new microbial ecosystem, by the antibacterial in microflora or
Funguses are under some natural endowments and non-existent by the degraded of the chemical substances such as synthetic insecticide, antibacterial, herbicide；Two
Being microorganism to be allowed directly using pesticide or antibacterial as growth substrate, and in the presence of multiple-microorganism group, makes pesticide
Change with the structure of antibiotic residues, so as to cause its chemical and physical features to change, that is, pass through pesticide or
Antibiotic residues are degraded to micromolecular compound from macromolecular compound, finally become H20 and C02Which is resolved into completely no
Machine thing, realizes being completely degraded, and becomes nontoxic inorganic matters；Three are degraded agriculture using microorganism enzymatic route and non-enzymatic route
Medicine,.Enzymatic reaction is enzyme system gene of the microorganism containing the degradable pesticide itself, by oxidation, dehydrogenation, reduction, hydrolysis,
The effect such as synthesis directly acts on pesticide, or, although the enzyme system without the pesticide of degrading, but under pesticide stress, microorganism
Gene there is restructuring or change, produce new degradation enzyme system, non-enzymatic reaction refers to that microbial activitiess occur the pH of environment
Change and cause degradation of pesticide, or produce the conversion that cofactor or chemical substance participate in pesticide.
Specifically used method and application one：The first step, to needing the agricultural environment that repairs to carry out pollutant component detection, surveys
Determine the carbon of soil, nitrogen, phosphorus, potassium content, determine the carbon source amount and ratio for needing, carbon source is with organic matter as standard；Second step, needs
C/N is adjusted to 25-30 between 6.2～20.9 by the agricultural soil C/N luffing of reparation；Per mu of rule uses 300-
500 kilograms of Organic substances with harmless treatment as carbon source, mix according to Organic substance weight add 10% organic nitrogen, 5% organic potassium,
5% organophosphors, trace element 0.1%；3rd step, according to 500-1000 gram of complex micro organism fungicide of per mu of administration, by composite microbial
Thing microbial inoculum is used according to 300-500 times of dilution proportion, the method after being mixed with the Organic substance of harmless treatment according to conventional fertilizer application.
Complex micro organism fungicide rehabilitating soil is the microbial bacteria under appropriate moisture, temperature, pH value condition, in microbial inoculum
Group is collectively forming dominant microflora with original beneficial microbe in soil, promotes the element such as carbon, nitrogen, oxygen in soil ecosystem
Benign cycle, degrading pesticide residues, so as to rehabilitating soil Eco-Environment System, make ecosystem reach new stable balance.
Wherein, after microbial composite bacteria system to the repair mechanisms of inorganic pollution is microbial manure entrance soil ecosystem, aerobic
The microorganism probiotic group such as bacterium, anaerobe, by the biological respinse of itself, reduces the acidity of soil, improves the pH value of soil, from
And reduce the murder by poisoning of harmful heavy metal in soil；While heavy metal can be fixed by the microbial bacteria in fertilizer, promote in soil
Active Heavy Metals are changed into organically combine state, form filter layer and sealing coat, reduce absorption of the crop to heavy metal in soil.According to
This method is implemented, and the degradation of pesticide rate for remaining in soil can reach more than 95%, the probability that heavy metal in soil is absorbed by plants
It is reduced to 3% or so.
Specifically used method and application two：
1st, the application in pollution by pesticides area is planted in Oryza sativa L., water root crop：Using microorganism compound repair 200 g/ mu of microbial inoculum/time,
500-800 times of dilution, transplanting, seedling throwing field, while soak field after mixing homogeneously with fertilizer or farm manure etc.；Live field, two leaves
Use when wholeheartedly, after mix broadcast application or dilution, inject flowing water mouth.Topdress（Before heading）200 g/ mus/time, coordinating topdresses applies
With, this product dilute with water is put into flowing water mouth, with current inflow Tanaka, or directly mix broadcast application.
2nd, the application in pollution by pesticides area is planted in field crop：Using microorganism compound repair 400～800 g/ mu of microbial inoculum/
Secondary, 300-500 times is diluted, coordinates fertilizer, farm manure etc. to use, by multiple dilutions, based on pouring root, supplemented by spray leaf, keep native
Push up moistening；Or directly mix liquid manure, rush fertilising pouring root, drip irrigation.
3rd, the application in pollution by pesticides area is planted in industrial crops：Using microorganism compound repair microbial inoculum 800～1600g/ mu/
Secondary, 300-500 times is diluted, coordinates fertilizer, farm manure etc. to use, by multiple dilutions, based on pouring root, supplemented by spray leaf, keep native
Push up moistening；Or directly mix liquid manure, rush fertilising pouring root, drip irrigation.Microbial inoculum is to the work in terms of the organic pollution reparation such as Pesticide Residue in Soil
With the preventing and treating number of times for being reduction pest and disease damage, the pesticide residues for having remained in soil of degrading, reduce the usage amount of pesticide, so as to
Reduce residual quantity of the pesticide in crop.Microbial composite bacteria can change plough horizon microbiota, in crop root week
Enclose dominant colony is formed, breed so as to strong inhibition pathogen, reduce pest and disease damage frequency；Meanwhile, microorganism is in its life
Many kinds of substance, such as hormoness, humic-acid kind and antibioticses is produced in active procedure, and these materials can stimulate plant growth to be good for
Strong, strengthen crop itself disease resistance ability.
Claims (15)
1. a kind of degraded antibiotic and pesticide residues complex micro organism fungicide, it is characterised in that：The complex micro organism fungicide is
By bacterial screening and training systern, actication of culture, amplification culture, dehydrate step, bacillus subtilis are obtained respectively
Bacterium, Bacillus licheniformis, aspergillus niger, colloid bacillus cereus, streptococcus thermophiluss, enterococcus faecalis, Lactobacillus plantarum, many viscous spore bars
Bacterium, bacillus megaterium, the thalline dry powder of 10 kinds of strains of pseudomonas fluorescens, the thalline dry powder blend of 10 kinds of strains is uniform,
The many thalline composite bacterium powder for obtaining.
2. complex micro organism fungicide as claimed in claim 1, it is characterised in that：Its preparation method is comprised the following steps：
（1）By bacillus subtilises, Bacillus licheniformis, aspergillus niger, colloid bacillus cereus, thermophilic coccus, enterococcus faecalis, plant
Lactobacilluss, bacillus polymyxa, bacillus megaterium, 10 kinds of bacterial strains of pseudomonas fluorescens carry out screening and the culture of bacterial strain respectively
Condition optimizing, obtains purification bacterial strain；Purification bacterial strain is carried out High Density Cultivation respectively, is inoculated into activation culture in shaking flask, will live
Change cultured strain be inoculated into fermentation tank be enlarged culture；After amplification culture terminates, list that amplification culture respectively is obtained
The zymocyte liquid of strain obtains the thalline dry powder of above-mentioned 10 kinds of strains through dehydrate；
（2）The thalline dry powder of 10 kinds of bacterial strains is uniformly mixed, obtains many thalline composite bacterium powder；In many thalline composite bacterium powder,
The percentage by weight that each live body bacterium accounts for composite bacterium powder total viable count is as follows：Bacillus subtilises 10-15%, aspergillus niger 10-15%, glue
Matter bacillus cereuss 10-15%, enterococcus faecalis 10-15%, Bacillus licheniformis 8-12%, bacillus megaterium 8-12%, fluorescence are false single
Born of the same parents bacterium 8-12%, Lactobacillus plantarum 5-8%, bacillus polymyxa 5-8%, streptococcus thermophiluss 6-8%；Many thalline composite bacterium powder
In, effective active bacterial content is hundred million cfu/g of 200-500, moisture 5%；Filtrated air is passed through in many thalline composite bacterium powder
Packaging, obtains final product complex micro organism fungicide.
3. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Prepared by bacillus subtilis strain withered
The comprising the following steps that of careless bacillus cereuss thalline dry powder：
（1）The screening of bacillus subtilis strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.015-0.02g/L, complementary element：DDT
0.1-0.2g/L, pH 7.0-7.2, by MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, Sodium Chloride, glucose, DDT, agar are dissolved in
In 1000 mL distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basal medium
When 45 DEG C ~ 50 DEG C are cooled to, the complementary element for being dissolved in 2 mL sterile purified waters is added, with the basis culture still in melt and dissolved state
Base mixes, and is then returned in sterilizing flat board, culture medium thickness at least high 5mm on flat board, and the final pH of culture medium in temperature is
When 25 DEG C, it is 7.2-7.5, the solid medium flat board containing DDT is obtained, dark place is placed in, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus subtilises that ampoul tube is preserved make bacterium solution, coat the training of the solid containing DDT
On foster base flat board, shading culture 2-3 days under the conditions of being placed in 35 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing DDT, be placed in 35 DEG C -38 DEG C, 180-200r/
Under min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of DDT concentration, 37-39 DEG C
Lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of DDT；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 2-3%
Inoculum concentration, be transferred in the fluid medium containing DDT, 35 DEG C -37 DEG C, 220-250r/min shaken cultivation 1-3 days, finally
With degradation rate of the high effective liquid chromatography for measuring bacterial strain to DDT；
7）Screening is obtained to DDT degradation rate highest bacillus subtilises, is preserved using test tube slant as purification bacterial strain, is placed in
4 DEG C of storages of refrigerator；
（2）Bacillus subtilis strain is activated：By step 7）Bacillus subtilises that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Being inoculated in activation culture shaking flask, with shaking flask rotating speed as 160-200r/min, 26h-32h is cultivated at a temperature of 35-37 DEG C, obtain
Liquid fermentation seed；The activation culture based formulas of above-mentioned activation culture are：Peptone 9.25-11.25g/L, Carnis Bovis seu Bubali cream 1.25-
1.3g/L、NaCl32.5-5.5g/L, glucose 22-25g/L, nutritional solution pH7.5-7.8；
（3）Bacillus subtilises amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 2-5%；The culture medium prescription of amplification culture is：Semen Maydis powder 18-20g/L,
Semen Glycines powder 20-22g/L, fish flour 5-6g/L, K2HPO41.2-1.5g/L、MgSO4 .7H2O 0.15-0.2g/L, Calcium Carbonate 6.5-
8.0g/L, ammonium sulfate 1.5-1.65g/L；In fermentation tank, temperature control is at 37-40 DEG C, and ferment pH7-8, and mixing speed is 200-
250r/min, fermentation time 28-32h；When the bacillus subtilis bacterial content in fermentation tank reaches 1010During individual/ml, will ferment
Bacillus subtilises bacterium solution drying dehydration be prepared into bacillus subtilises thalline dry powder.
4. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：By Bacillus licheniformis（Bacillus
licheniformis）Bacterial strain prepares comprising the following steps that for Bacillus licheniformis soma powder：
（1）The screening of lichem bacillus strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：MnSO40.5-1.0g/L, peptone 8-10g/L, cattle
Meat extract 2-3g/L, NaCl 3-5g/L, glucose 1.5-2.5g/L, agar 15-20g/L, complementary element：Bayer 71628 0.15-
0.25g/L, pH7.0-7.2, by MnSO4, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, Bayer 71628, agar be dissolved in 1000 mL
In distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled to
When 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2 mL sterile purified waters is added, mix with the basal medium for being still in melt and dissolved state
Close, be poured in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium, when temperature is 25 DEG C, is 7.0-
7.2, the solid medium flat board containing Bayer 71628 is obtained, dark place is placed in, preserve in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Bacillus licheniformis that ampoul tube is preserved make bacterium solution, coat consolidating containing Bayer 71628
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 30 DEG C -32 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing Bayer 71628, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing Bayer 71628, be placed in 30 DEG C -32 DEG C, 160-
Under 180r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of methylamine phosphorus concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate is as capable of the bacterial strain of degrading methamidophos；
6）By step 5）It is capable of the bacterial strain of degrading methamidophos, is transferred on activation medium respectively and is activated；Then will activation
Bacterium solution press the inoculum concentration of 3%-5%, be transferred in the fluid medium containing Bayer 71628,30 DEG C -32 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to Bayer 71628；
7）Screening is obtained to methamidophos degradation rate highest lichem bacillus strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus licheniformis strain is activated：By step 7）Bacillus licheniformis that screening is obtained, being placed in 4 DEG C of storages of refrigerator
Slant strains are inoculated in activation culture shaking flask, with shaking flask rotating speed 160r/min, cultivate 16h-20h, obtain at a temperature of 28-30 DEG C
Obtain liquid fermentation seed；The activation culture based formulas of the activation culture are：Peptone 9.25-10.0g/L, Carnis Bovis seu Bubali cream 4.5-
5g/L, NaCl4.8-5.0g/L, glucose 1.5g-2.0/L, nutritional solution pH6.5-7.2；
（3）Bacillus licheniformis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 3-5%；Amplification culture based formulas are：Semen Maydis powder 15-18g/L, Semen Glycines powder
22-25g/L、K2HPO40.08-0.20g/L、MgSO4 .7H2O 0.005-0.01g/L, yeast powder 0.1-0.15g/L；Fermentation temperature
25-40 DEG C of degree, ferment pH value 6-8, and it is 8-10%, fermentation time 18- that mixing speed is 200-300r/min, air mass flow ratio
28h；When the Bacillus licheniformis content in fermentation tank reaches 1010During individual/ml, by the Bacillus licheniformis liquid warp for fermenting
Drying and dehydrating is prepared into Bacillus licheniformis soma powder.
5. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Aspergillus niger is prepared by Aspergillus niger strain
The comprising the following steps that of soma powder：
（1）The screening of Aspergillus niger strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 5-10g/L, glucose 20-25g/L, sulfur
Sour magnesium 0.5-1.0g/L, yeast extract powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1.0-2.0g/L, agar 15-20g/L, mend
Fill composition：Glyphosate 0.15-0.25g/L, N,N'-dimethyl-.gamma..gamma.'-dipyridylium 0.2-0.5g/L, oxytetracycline 0.1-0.2g/L, by peptone, glucose,
Magnesium sulfate, yeast extract powder, dipotassium hydrogen phosphate, agar are dissolved in 1000 mL distilled water, are subsequently adding 10 mL glycerol, plus
Heat is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C,
Add and the complementary element glyphosate of 2mL sterile purified water, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, oxytetracycline is dissolved in, with the basis training still in melt and dissolved state
Foster base mixing, is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium when temperature is 25 DEG C is
6.5-6.8, obtains the solid medium flat board containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, is placed in dark place, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common Aspergillus niger that ampoul tube is preserved makes bacterium solution, coats containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
On solid medium flat board, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated each bacterium colony, in the flat lining out of the solid medium containing glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, enters one
Step is separated and purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing in glyphosate, the fluid medium of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be placed in 30 DEG C -32 DEG C
Under 120-150r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, draws on the different inorganic salt solid medium flat board of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium concentration
Line, 30-32 DEG C of lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate as being capable of degradation of glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Bacterial strain；
6）By step 5）Obtain can degradation of glyphosate, the bacterial strain of N,N'-dimethyl-.gamma..gamma.'-dipyridylium, be transferred on activation medium respectively and activated；
Then the bacterium solution of activation is pressed the inoculum concentration of 1-3%, is transferred to glyphosate, in N,N'-dimethyl-.gamma..gamma.'-dipyridylium fluid medium, 25 DEG C -28 DEG C,
160-180r/min vibration light culture 1-3 days, finally with high effective liquid chromatography for measuring bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium
Degradation rate；
7）Screening obtains the degradation rate highest Aspergillus niger bacterial strain to herbicide glyphosate, N,N'-dimethyl-.gamma..gamma.'-dipyridylium, adopts as purification bacterial strain
Preserved with test tube slant, be placed in 4 DEG C of storages of refrigerator；
（2）Aspergillus niger strain is activated：By step 7）The aspergillus nigers for being placed in 4 DEG C of storages of refrigerator are inoculated into work in activation culture shaking flask
Change, activation culture based formulas are peptone 5.0-5.5g/L, glucose 10.0-15g/L, magnesium sulfate 0.5-0.8g/L, yeast
Powder 2.0-3.0g/L, dipotassium hydrogen phosphate 1-3g/L, nutritional solution pH7.0-7.2 is leached, shaking flask rotating speed is 100-160r/min,
25h-28h is cultivated at a temperature of 28-35 DEG C, obtain liquid fermentation seed；
（3）Aspergillus niger amplification culture, dehydrate：By step（2）Obtain the fermentation that liquid fermentation seed is inoculated into amplification culture
Ferment in tank, be enlarged culture, inoculum concentration 8-10%；Amplification culture based formulas Rhizoma Solani tuber osi 35-40g/L, glucose 20-50g/
L, dipotassium hydrogen phosphate 5-8g/L, magnesium sulfate 1-1.5g/L, pH value are 7.0；Fermentation temperature is 35 DEG C, mixing speed 100-180r/
Min, fermentation time 25-28h, when the aspergillus niger content in fermentation tank reaches 1010During individual/ml, by the Aspergillus niger for fermenting
The dehydration of liquid drying is prepared into aspergillus strain dry powder.
6. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：By colloid bacillus cereus（Bacillus
mucilaginosus Krassilnikov）Bacterial strain prepares comprising the following steps that for colloid bacillus cereus thalline dry powder：
（1）The screening of colloid bacillus cereus bacterial strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 2-3g/L, ammonium sulfate 0.15-0.25g/L,
Yeast extract 0.1-0.g/L, KCl 0.01-0.02g/L, MgSO4 .7H2O 0.01-0.02g/L、NaH2PO40.01-0.02g/L、
CaCO30.1-0.2g/L, silica 1-2g/L, agar 10-15g/L, complementary element：Cypermethrin 0.1-0.3g/L,
CuSO4 .5H2O 0.02-0.025g/L, initial pH 7.0-7.2；By glucose, ammonium sulfate, yeast extract, KCl, MgSO4 .7H2O、
NaH2PO4、CaCO3, silicon dioxide, CuSO4 .5H2O, agar are dissolved in 1000mL distilled water, heating boil molten, and at 121 DEG C
High pressure steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL sterilizing
The complementary element cypermethrin of distilled water, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, training
Foster base thickness at least high 5mm, the final pH of culture medium is 7.0-7.5, must have cypermethrin and copper sulfate when temperature is 25 DEG C
Solid medium flat board, be placed in dark place, in 3 DEG C ~ 6 DEG C preserve；
2）Inoculation and purification：The common colloid bacillus cereus that ampoul tube is preserved make bacterium solution, coat containing cypermethrin and
On the solid medium flat board of copper sulfate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of cypermethrin concentration, 32-
35 DEG C of lucifuge cultures 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of cypermethrin；
6）By step 5）Obtain degrading the bacterial strain of cypermethrin, is transferred on activation medium respectively and is activated；Then
The bacterium solution of activation is pressed the inoculum concentration of 2-5%, has been transferred in cypermethrin fluid medium, 37 DEG C -40 DEG C, 200-220r/
Min shaken cultivation 1-3 days, finally uses degradation rate of the high effective liquid chromatography for measuring bacterial strain to cypermethrin；
7）Screening obtains the degradation rate highest colloid bacillus cereus bacterial strain to cypermethrin, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Colloid bacillus cereus actication of culture：By step 7）Colloid bacillus cereus inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Starch 15-20g/L, dipotassium hydrogen phosphate g/L, ammonium sulfate 2-3g/
L, yeast extract 3.5-5.5g/L, sucrose 8.5-10g/L, magnesium sulfate 4.5-5g/L, Calcium Carbonate 2.5-3g/L, silicon dioxide 6.5-
8g/L, ferric chloride 0.05-lg/L, pH7.0-7.2, shaking flask rotating speed is to cultivate at a temperature of 30-35 DEG C
18-20h, obtains liquid fermentation seed；
（3）Colloid bacillus cereus amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration be 4-6%；The composition of fermentation medium is sucrose 2.5-6.5gg/L, phosphorus
Sour hydrogen dipotassium 3-5.5gg/L, yeast extract 3-6g/L, ammonium sulfate 2.5-5gg/L, soy peptone 5-10g/L, magnesium sulfate 3-6/
g/L、pH7.2-7.5；Amplification culture condition：32 DEG C of fermentation temperature, pH7.2, mixing speed 180r/min-220r/min, fermentation
Time 24h；When the colloid bacillus cereus content of fermentation tank reaches 1010During individual/ml, by the colloid bacillus cereus bacterium solution for fermenting
Drying dehydration is prepared into colloid bacillus cereus thalline dry powder.
7. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Prepared by strains of streptococcus thermophilus thermophilic
The comprising the following steps that of streptococcus thalline dry powder：
（1）The screening of strains of streptococcus thermophilus is carried out according to a conventional method with training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 2-3g/L, peptone 2-3g/
L, yeast extract 1-2g/L, Fructus Lycopersici esculenti juice 15-20g/L, glucose 2-4g/L, tween 0.5ml-1.0ml/L, Calcium Carbonate 1.0-2.0g/
L, agar 2-5g/L, pH6.5-7.0, take suitable quantity of water in pot according to ratio, heating, by Carnis Bovis seu Bubali cream, peptone, yeast extract, kind
Tomato juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour in the conical flask for added agar and add after medicine boiling in pot
Plug, wrapping, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator culture 24h, asepsis growth person
Can use；The culture medium using when can also make fluid medium without agar, for the shaken cultivation of mushroom；
The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common streptococcus thermophiluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, culture obtains bacterial strain；
6）By step 5）Each bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 5%-
8% inoculum concentration, is transferred in fluid medium, 37 DEG C, quiescent culture 1-2 days, determines the biological active matter produced by each bacterial strain
Matter content；
7）Screening obtains producing bioactive substance content highest strains of streptococcus thermophilus, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Streptococcus thermophilus strain is activated：By step 7）Streptococcus thermophiluss obtaining, being placed in 4 DEG C of storages of refrigerator are inoculated into work
Change activation culture in culture shaking flask, activation culture based formulas are：Tryptone 1-3.0g/L, fish peptone 2-3g/L, Carnis Bovis seu Bubali cream
2-3g/L, yeast extract 0.5-1.5g/L, magnesium sulfate 0.3-0. 5g/L, potassium dihydrogen phosphate 0.5-1.0g/L, Lactose 1-1.5g/L,
Nutritional solution pH 6.5-6.8, cultivates 24h-28h at a temperature of 35-40 DEG C, and mixing speed is 80-100r/min, obtains liquid and sends out
Ferment seed；
（3）Streptococcus thermophiluss amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 2-5%；Amplification culture based formulas：Glucose 1-3g/L, soy peptone 4-
6g/L, dipotassium hydrogen phosphate 2-3g/L, corn juice 40-50g/L, sodium acetate 1-2g/L, skimmed milk 10-15g/L, pH are 6.5-6.8；
38-45 DEG C of fermentation temperature, mixing speed is 100-120r/min, fermentation time 48-72h；When the streptococcus thermophiluss in fermentation tank
Content reaches 109Lyophilizing after bacterium solution centrifugation, prepared streptococcus thermophiluss thalline dry powder will be cultivated during individual/ml.
8. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Excrement intestinal ball is prepared by E. Faecium strains
The comprising the following steps that of bacterium thalline dry powder：
（1）The screening of E. Faecium strains and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Peptone 10-15g/L, maltose 20-25g/L, ferment
Female leaching powder 10-2g/L, sodium glycerophosphate 5-6g/L, Sodium Chloride 1-2g/L, Lactose 0.015-0.020g/L, agar 15-20, pH
Value 8.0-8.5, is separately separately added into complementary element in order in the medium：Erythromycin 0.05-0.15g/L, ciprofloxacin
0.01-0.02g/L, Sulfamethoxazole Compound 0.25-0.35g/L；By peptone, maltose, yeast extract, sodium glycerophosphate,
Sodium Chloride, Lactose, agar are dissolved in 1000mL distilled water, are subsequently adding 10mL glycerol, heating boil molten, and in 121 DEG C of high pressure
Steam sterilization 15min, obtains basal medium, when basal medium is cooled to 45 DEG C ~ 50 DEG C, adds and is dissolved in 2mL aseptic distillation
The complementary element erythromycin 0.05-0.15g/L of water, ciprofloxacin 0.01-0.02g/L, Sulfamethoxazole Compound 0.25-
0.35g/L, is mixed with the basal medium still in melt and dissolved state, is poured in sterilizing flat board, and culture medium thickness are at least high
5mm, the final pH of culture medium, when temperature is 25 DEG C, is 7.5-8.0, dislike containing erythromycin, ciprofloxacin, compound sulfonamide first
Azoles solid medium flat board, is placed in dark place, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common enterococcus faecalis that ampoul tube is preserved make bacterium solution, coat husky containing erythromycin, ring third
On star, Sulfamethoxazole Compound solid medium flat board, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium
Fall；
3）By step 2）Isolated bacterium colony, containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound solid medium
Flat lining out, is separated and purification further；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed into containing erythromycin, ciprofloxacin, Sulfamethoxazole Compound liquid culture
In base, 33 DEG C -35 DEG C are placed in, under 160-180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, solid in the different inorganic salt of erythromycin, ciprofloxacin, Sulfamethoxazole Compound concentration
Rule on body culture medium flat plate, 35 DEG C -38 DEG C lucifuge cultures 2-5 days；The bacterial strain that can grow on culture medium flat plate is energy
Enough degraded erythromycin, ciprofloxacin, the bacterial strain of Sulfamethoxazole Compound；
6）By step 5）Obtain degrading the bacterial strain of erythromycin, ciprofloxacin, Sulfamethoxazole Compound, is transferred to activation respectively
Activated in culture medium, then the bacterium solution of activation pressed the inoculum concentration of 3%-5%, be transferred to pH8.5-9.0 containing erythromycin,
In ciprofloxacin, Sulfamethoxazole Compound element fluid medium, at 38 DEG C -40 DEG C, 180-250r/min vibrates light culture 1-3
My god, finally with each bacterial strain of high effective liquid chromatography for measuring to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate；
7）Screening obtain to erythromycin, ciprofloxacin, Sulfamethoxazole Compound degradation rate highest E. Faecium strains, as
Purification bacterial strain is preserved using test tube slant, is placed in 4 DEG C of storages of refrigerator；
（2）Enterococcus faecalis actication of culture：By step 7）The enterococcus faecalis for being placed in 4 DEG C of storages of refrigerator for obtaining are inoculated into activation culture
Activate in shaking flask, activation culture based formulas are：Glucose 4-5g/L, yeast powder 0.15-0.3g/L, peptone 0.5-1g/L, no
Water calcium chloride 0.002-0.006g/L, bitter salt 0.02-0.05g/L, dipotassium hydrogen phosphate 0.1-0.15g/L, seven hydrations
Manganese sulfate 0.002-0.005g/L, Calcium Carbonate 0.5-1g/L, nutritional solution pH7.5-8.0, shaking flask rotating speed is 180-200r/min,
30h-35h is cultivated at a temperature of 30-35 DEG C, obtain liquid fermentation seed；
（3）Enterococcus faecalis amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into sending out for amplification culture
Ferment in fermentation tank, be enlarged culture, inoculum concentration 8-10%；Culture medium prescription is tryptone 10-15g/L, yeast powder 10-
15g/L, glucose 10-15g/L, anhydrous calcium chloride 0.04-0.06g/L, bitter salt 0.019-0.025g/L, phosphoric acid hydrogen
Dipotassium 0.04-0.06g/L, 0.04-0.06g/L of potassium dihydrogen phosphate, dibastic sodium phosphate 0.4-0.6g/L, Sodium Chloride 0.08-0.1g/
L, murphy juice 50-80g/L, Calcium Carbonate 1-3g/L, sodium acetate 10-15g/L, ammonium carbonate 2-3g/L, ammonium acetate 1-2g/L, half Guang ammonia
Acid hydrochloride 0.5-1g/L, pH value is 8.5-9；35-37 DEG C of fermentation temperature, mixing speed 180-220r/min, fermentation time 20-
32h, when the enterococcus faecalis content of fermentation tank reaches 1010During individual/ml, by the enterococcus faecalis bacterium solution drying for fermenting dehydration system
Standby become enterococcus faecalis thalline dry powder.
9. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Plant is prepared by lactobacillus plantarum strain
The comprising the following steps that of lactobacilluss thalline dry powder：
（1）The screening of lactobacillus plantarum strain and training systern, are carried out according to a conventional method：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Water 100ml, Carnis Bovis seu Bubali cream 1-2g/L, peptone 1-2g/
L, yeast extract 1-2g/L, Fructus Lycopersici esculenti juice 20-30g/L, glucose 1-2g/L, tween 0.05ml-0.10ml/L, Calcium Carbonate 1.5-
2.0g/L, agar 2-5g/L, pH6.2-6.5, according to ratio according to the required suitable quantity of water that measures in pot, heating, weigh Carnis Bovis seu Bubali cream,
Peptone, yeast extract, Fructus Lycopersici esculenti juice, glucose, tween, Calcium Carbonate are sequentially added in water, pour into and add after medicine boiling in pot
Jump a queue in the conical flask of agar, wrap up, 121 DEG C of sterilizing 20min, the culture medium after must sterilizing, it is placed on 37 DEG C of constant incubator trainings
Foster 24h, asepsis growth person can use；Culture medium using when can also make fluid medium without agar, for bacterium
The shaken cultivation of class；The Fructus Lycopersici esculenti juice can be formed using fresh Fructus Lycopersici esculenti squeezing；
2）Inoculation and purification：The common plant lactobacilluss that ampoul tube is preserved make bacterium solution, coat on solid medium flat board,
Shading culture 2-3 days under the conditions of being placed in 37 DEG C -38 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be statically placed in 36 DEG C -38 DEG C, cultivate 1-3 days, obtain list
One bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 37-38 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）The each bacterial strain for obtaining, is transferred on activation medium respectively and is activated, and then presses the bacterium solution of activation
The inoculum concentration of 3%-5%, is transferred in fluid medium, 37 DEG C, and quiescent culture 1-2 days finally determines the life produced by each bacterial strain
Active substances content；
7）Screening obtains producing bioactive substance content highest lactobacillus plantarum strain, oblique using test tube as purification bacterial strain
Face preserves, and is placed in 4 DEG C of storages of refrigerator；
（2）Lactobacillus plantarum actication of culture：By step 7）Lactobacillus plantarum obtaining, being placed in 4 DEG C of storages of refrigerator is inoculated into work
Change and activate in culture shaking flask, activation culture based formulas are：Peptone 10-12g/L, Carnis Bovis seu Bubali cream 10-15g/L, yeast extract 5-6.5g/
L、KH2PO42-3.5g/L, trisodium citrate 2-2.5g/L, sodium acetate 2-2.5g/L, glucose 20-28g/L, MgSO4 .7H2O
0.58-0.60g/L、MnSO4 .4H2O 0.25-0.3g/L, nutritional solution pH 6.2-6.5, cultivate 24h- at a temperature of 35-40 DEG C
28h, quiescent culture, obtain liquid fermentation seed；
（3）Lactobacillus plantarum amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, is enlarged culture, inoculum concentration 5-8%；Culture medium prescription is：Sucrose 25-30g/L, yeast extract 15-20g/
L、KH2PO48-10g/L, trisodium citrate 5-6g/L, sodium acetate 3-5g/L, MgSO4 .7H2O 0.8 -1.2g/L、MnSO4 .4H2O
0.5-0.8g/L, pH 6.2-6.5；35-37 DEG C of fermentation temperature, fermentation time 24-28h, quiescent culture；Fermentation ends, will fermentation
Good Lactobacillus plantarum bacterium solution drying dehydration is prepared into Lactobacillus plantarum thalline dry powder.
10. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Prepared by bacillus polymyxa bacterial strain many
The comprising the following steps that of viscous bacillus cereuss thalline dry powder：
（1）The screening of bacillus polymyxa bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-15g/L, yeast extract 5-8g/
L、NaCl 3-5g/L、KH2PO40.1-0.2g/L、MgSO4 .7H2O 0.05-0.1g/L、MnSO4 .7H2O 0.25-0.5g/L, fine jade
Fat 15-20g/L, pH7.5-8.0；By MnSO4 .7H2O, peptone, KH2PO4、NaCl、MgSO4 .7H2O, agar are dissolved in 1000
In mL distilled water, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats that basal medium is cooled down
During to 45 DEG C ~ 50 DEG C, it is poured in sterilizing flat board, culture medium thickness at least high 5mm, the final pH of culture medium is 25 DEG C in temperature
When, it is 7.5-8.0, solid medium flat board is obtained, dark place is placed in, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common bacillus polymyxa that ampoul tube is preserved makes bacterium solution, coats solid medium flat board
On, shading culture 2-3 days under the conditions of being placed in 37 DEG C -40 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of solid medium, is separated and purification further；This step weight
Multiple 3-5 time, until obtaining single dominant bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in fluid medium, be placed in 38 DEG C -40 DEG C, 160-180r/min vibrate bar
Under part, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of solid medium, 38-40 DEG C of lucifuge culture 2-3 days；In solid
On culture medium flat plate, growth obtains bacterial strain；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated, then the bacterium solution of activation is pressed 2-5%
Inoculum concentration, be transferred in fluid medium, 37 DEG C -40 DEG C, 180-200r/min shaken cultivation 1-3 days, finally determine each bacterium
The various bioactivators contents such as such as antibiotic, antagonist protein, phytohormone, enzyme, flocculant produced by strain；
7）Screening obtains producing bioactive substance content highest bacillus polymyxa bacterial strain, adopts test tube as purification bacterial strain
Inclined-plane is preserved, and is placed in 4 DEG C of storages of refrigerator；
（2）Bacillus polymyxa actication of culture：By step 7）Screening obtains, is placed in the bacillus polymyxa of 4 DEG C of storages of refrigerator and connects
Plant and activate in activation culture shaking flask, activation culture based formulas are：Peptone 8-10g/L, Carnis Bovis seu Bubali cream 3-5g/L, Sodium Chloride 3-
5g/L, glucose 10-12g/L, KH2PO40.2-0.3g/L、MgSO4 .7H2O 0.2-0.35g/L、NaCl 0.2-0.3g/L、
CaSO4 .2H2O 0.2-0.25g/L、CaCO35.0-6.5g/L, nutritional solution pH 7.0-8.0, shaking flask rotating speed 150-170r/min,
25h-30h is cultivated at a temperature of 37-40 DEG C, obtain liquid fermentation seed；
（3）Bacillus polymyxa amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 10-12%；Amplification culture based formulas are：Semen Maydis powder 20-25g/L, Semen Glycines powder
8-12g/L、K2HPO40.05-0.15g/L, Sodium Chloride 1.0-1.5g/L, Na2HP040.15-0.25g/L；Fermentation temperature 28-
35 DEG C, ferment pH7-8, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h, and mixing speed is 160-200r/min, fermentation time 20-25h；When the bacillus polymyxa of fermentation tank contains
Amount reaches 1010During individual/ml, the bacillus polymyxa bacterium solution drying for fermenting dehydration is prepared into bacillus polymyxa thalline
Dry powder.
11. complex micro organism fungicide as claimed in claim 1 or 2, it is characterised in that：Prepared by bacillus megaterium bacterial strain huge
The comprising the following steps that of Bacterium anthracoides thalline dry powder：
（1）The screening of bacillus megaterium bacterial strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Glucose 18-20g/L, peptone 15-18g/L, chlorine
Change sodium 5-6g/L, Carnis Bovis seu Bubali cream 0.5-1g/L, agar 15-20g/L, MnSO4 .7H2O 0.025-0.035g/L, complementary element：To sulfur
Phosphorus 0.15-0.20g/L, pH7.5-8.0,；By MnSO4 .7H2O, peptone, Carnis Bovis seu Bubali cream, NaCl, glucose, parathion, agar are molten
Solution is in 1000 mL distilled water, and heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basis
When culture medium is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the base still in melt and dissolved state
Basal culture medium mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is 25 DEG C in temperature
When be 7.5-8.0, obtain the solid medium flat board containing parathion, be placed in dark place, in 3 DEG C ~ 6 DEG C preserve；
2）Inoculation and purification：The common bacillus megaterium that ampoul tube is preserved makes bacterium solution, coats consolidating containing parathion
On body culture medium flat plate, shading culture 2-3 days under the conditions of being placed in 28 DEG C -30 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing parathion, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing parathion, be placed in 28 DEG C -30 DEG C, 180-
Under 200r/min oscillating condition, cultivate 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of parathion concentration, 30-32
DEG C lucifuge culture 2-3 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of parathion；
6）By step 5）Obtain degrading the bacterial strain of parathion, is transferred on activation medium respectively and is activated, then will
The bacterium solution of activation presses the inoculum concentration of 2-5%, has been transferred in parathion fluid medium, and 37 DEG C -40 DEG C, 200-220r/min shakes
Culture 1-3 days is swung, finally the degradation rate with each bacterial strain of high effective liquid chromatography for measuring to parathion；
7）Screening obtains the degradation rate highest bacillus megaterium bacterial strain of parathion, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Bacillus megaterium actication of culture：By step 7）Bacillus megaterium inoculation obtaining, being placed in 4 DEG C of storages of refrigerator
Activate in activation culture shaking flask, activation culture based formulas are：Peptone 10-15g/L, Carnis Bovis seu Bubali cream 3-5g/L, NaCl6.5-
8.5g/L, glucose 2.5-3g/L, nutritional solution pH7.0-7.2, shaking flask rotating speed is to cultivate 190-210r/min, at 37-40 DEG C
22-28h, obtains liquid fermentation seed；
（3）Bacillus megaterium amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5-9%；The culture medium prescription of amplification culture is：Semen Maydis powder 15-18g/L,
Semen Glycines powder 25-30g/L, fish flour 5-6g/L, K2HPO40.05-0.15g/L, calcium chloride 5.5-6.5.0g/L, ammonium sulfate 2.0-2.5g/
L；36-40 DEG C of fermentation temperature, fermentation pH value issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss 7-8, and fermentation tank mixing speed is 200-250r/min, fermentation time 25-32h issssssssssssssssssssssssssssssssssssssssss；
When the bacillus megaterium content in fermentation tank reaches 1010During individual/ml, by the bacillus megaterium bacterium solution drying for fermenting
Dehydration is prepared into bacillus megaterium thalline dry powder.
12. complex micro organism fungicides as claimed in claim 1 or 2, it is characterised in that：Prepared by pseudomonas fluorescens strain glimmering
The comprising the following steps that of light pseudomonass thalline dry powder：
（1）The screening of pseudomonas fluorescens strain and training systern：
1）Screening and culturing based formulas and preparation：By following culture medium prescription：Tryptone 10-12g/L, K2SO410-12g/L、
MgCl21.4-1.6g/L、Fe2(SO4)30.5-1g/L, 10 mL of glycerol, 15 ~ 20g/L of agar, 1 L of distilled water, supplement into
Point：Penicillin 0.05-0.10g/L；By tryptone, K2SO4、MgCl2、Fe2(SO4)3, agar be dissolved in 1000 mL distilled water
In, it is subsequently adding 10 mL glycerol, heating is boiled molten, and in 121 DEG C of high pressure steam sterilization 15min, obtains basal medium, treats basis
When culture medium is cooled to 45 DEG C ~ 50 DEG C, the complementary element for being dissolved in 2mL sterile purified water is added, with the base still in melt and dissolved state
Basal culture medium mixes, and is poured in sterilizing flat board, culture medium thickness at least high 5mm, and the final pH of culture medium is 25 DEG C in temperature
When, it is 7.0-7.2, the solid medium containing penicillin is obtained, dark place is placed in, preserves in 3 DEG C ~ 6 DEG C；
2）Inoculation and purification：The common fluorescent pseudomonass that ampoul tube is preserved make bacterium solution, coat consolidating containing penicillin
On body culture medium flat plate, shading culture 2-5 days under the conditions of being placed in 25 DEG C -28 DEG C, obtain different bacterium colonies；
3）By step 2）Isolated bacterium colony, in the flat lining out of the solid medium containing penicillin, separate further and
Purification；This step repeats 3-5 time, until obtaining single bacterium colony；
4）By step 3）Single bacterium colony be sequentially ingressed in the fluid medium containing penicillin, be placed in 25 DEG C -28 DEG C, 150-
Under 180r/min oscillating condition, shading culture 1-3 days, obtain single bacterium colony culture fluid；
5）By step 4）The culture fluid for obtaining, in the flat lining out of the different inorganic salt solid medium of penicillin concn, 25-28
DEG C lucifuge culture 2-5 days；The bacterial strain that can grow on culture medium flat plate can as be degraded the bacterial strain of penicillin；
6）By step 5）Bacterial strain is obtained, is transferred on activation medium respectively and is activated；Then the bacterium solution of activation is pressed 1%-3%
Inoculum concentration, be transferred to containing in penicillin fluid medium, at 25 DEG C -28 DEG C, 160-180r/min vibrates light culture 1-3
My god, finally use degradation rate of the high effective liquid chromatography for measuring bacterial strain to penicillin；
7）Screening is obtained to penicillin degradation rate highest pseudomonas fluorescens strain, is protected using test tube slant as purification bacterial strain
Deposit, be placed in 4 DEG C of storages of refrigerator；
（2）Pseudomonas fluorescens actication of culture：By step 7）The pseudomonas fluorescens inclined-plane bacterium for being placed in 4 DEG C of storages of refrigerator for obtaining
Plant to be inoculated in activation culture shaking flask and activate, activation culture based formulas are：Carnis Bovis seu Bubali cream 2.55-3.0g/L, peptone 1.65-
2.2g/L, Sodium Chloride 0.18g-0.2g/L, yeast extract 5.25-6.0g/L, Fe2(SO4)31-1.5g/L, culture fluid pH7.0-
7.5, shaking flask rotating speed is to cultivate 18-22h at a temperature of 28.5-31 DEG C, obtains liquid fermentation seed；
（3）Pseudomonas fluorescens amplification culture, dehydrate：By step（2）Obtain liquid fermentation seed and be inoculated into amplification culture
Fermentation cylinder for fermentation, be enlarged culture, inoculum concentration 5.5 ~ 8.5%；The culture medium prescription of the amplification culture is：Maltose
20-35g/L, Semen Maydis powder 12-15g/L, yeast extract 4.5-4.62g/L, ammonium sulfate 5.2-5.8g/L, K2HPO40.55-0.85g/
L、MgSO4 .7H2O 0.035-0.055g/L、FeSO4 .7H2O 1.095-1.20g/L、CaCl20.075-1.0g/L、NaCl
0.15-0.25g/L；27 DEG C ~ 29 DEG C of fermentation temperature, fermentation tank 140 ~ 180r/min of mixing speed, ventilation 8 ~ 12%, during fermentation
Between 24 ~ 48h；When the Pseudomonas bacterial content in fermentation tank reaches 1010During individual/ml, by the pseudomonas fluorescens for fermenting
The dehydration of bacterium solution drying is prepared into pseudomonas fluorescens thalline dry powder.
13. complex micro organism fungicides as claimed in claim 1 or 2, it is characterised in that：In organic pollution harmless treatment
In, be applied to carry out feces of livestock and poultry compost fermentation, to agriculture waste Organic substance, industrial processes Organic Ingredientss, organic waste, have
The harmless treatment of machine sludge.
14. complex micro organism fungicides as claimed in claim 1 or 2, it is characterised in that：In field of agricultural cultivation, improved soil,
Biological control, the application for reducing in agriculture border pollution, increase crop yield and raising crop product quality.
15. complex micro organism fungicides as claimed in claim 1 or 2, it is characterised in that：As agricultural environment renovation agent, in drop
Solution pesticide and the application for processing in heavy metal pollution.
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