CN106906170A - Complex micro organism fungicide and its preparation method and application - Google Patents
Complex micro organism fungicide and its preparation method and application Download PDFInfo
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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
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/341—Consortia of bacteria
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/007—Contaminated open waterways, rivers, lakes or ponds
Abstract
The invention discloses a kind of complex micro organism fungicide and its preparation method and application, the complex micro organism fungicide includes pseudomonad preparation, acinetobacter calcoaceticus preparation, bacillus amyloliquefaciens preparation, Ko subtilis bar preparation, bacillus licheniformis preparation, Lactococcus lactis bacteria preparation, hydrogenlike silicon ion preparation, alkali lake enlightening thatch Salmonella preparation and Rhodopseudomonas palustris preparation.Preparation method is comprised the following steps：(1) complex micro organism fungicide component is prepared；(2) complex micro organism fungicide component is mixed in proportion, obtains complex micro organism fungicide.The complex micro organism fungicide can simultaneously decompose the growth of phosphorus element concentration and suppression algae in ammonia nitrogen and nitrate, reduction water body in different organic matters, degradation water.Carry out town and country river sewage in-situ immobilization using complex micro organism fungicide of the invention, with efficient, quick, non-secondary pollution, it is simple to operate the features such as, be conducive to large-scale promotion application.
Description
Technical field
The invention belongs to technical field of environmental microorganism, it is related to a kind of for the compound micro- of town and country river sewage in-situ immobilization
Bacteria agent and its preparation method and application.
Background technology
City river and lake be with human being's production and the closely coupled water environment of living, it is important as urban ecological system
Part, with supply water source, improve environment, the functions, the life to city such as greenery patches, culture and education and amusement and leisure be provided
State is built significant.But with the continuous quickening of urbanization process, urban population aggregation, economic activity frequency increasingly
Numerous, industrial wastewater and sanitary sewage largely discharge increasing, and the random discarding of rubbish causes organic matter, ammonia nitrogen, total p and ses in water body
Change the pollutants such as hydrogen to continue to increase, problem of environmental pollution is increasingly serious, the serious society for restricting city, economy and continuity of environment
Development, while jeopardizing the healthy of the people.
The traditional means of removal water pollutant mainly have physics and chemical method, although administered in river course and lake pollution
It is upper that there is certain effect, but there is also problem.Can such as Sediment Dredging in physical method have length to pollution of river thing
The control of effect and the also presence dispute that whether had a negative impact to benthonic realm.Addition molysite in chemical flocculation treatment technology
Promote phosphorus precipitation, add the methods such as lime denitrogenation, although instant effect, efficiency high, but easily cause secondary pollution.
Bioremediation technology is the new technology that the water pollutant that recent fast development is got up is repaired, and it is using specific
Biological particularly microorganism is to the absorption of pollutant in water body, conversion or degrades, and reaches and slows down or finally eliminate water pollution, extensive
The biological control measure of rehydration body ecological functions.Microorganism tool is because its cultivation cycle is short, growth and breeding is rapid, adaptable, conversion
The features such as efficiency high, played an important role during polluted river water is administered.The work of microorganism remediation river sewage
With mainly including：1) microorganism is Self substances, another aspect microorganism by assimilation meeting transform portion organic pollution
Different biology enzyme energy fast degradation larger molecular organicses can be produced, degraded and removal to organic pollutants in water body is realized.2)
Some amonifying bacterias, nitrobacteria and denitrifying bacteria can realize Water element with the ammonia nitrogen and nitrate in degradation water
Biogeochemical cycle process, and finally make excessive nitrogen in water body that water body is escaped in the form of nitrogen.3) one slightly
Biology can reduce phosphorus element concentration in water body by the metabolism of characteristic, and another aspect certain micro-organisms can be turned into by same
With, the available phosphorus in water body is converted into the organophosphor of thalline itself, acted on by the transmission of food chain, in removal water body
Phosphorus.4) by the synergy of complex microorganism, water body towel nitrogen, phosphorus recycling level are reduced, is pressed down by way of nutrient competition
The growth of algae processed；Simultaneously by improving water body physicochemical environment, promote the growth of zooplankter population to grow, caught with zooplankter
The mode for eating algae controls the growth of algae.
The patent of invention of Application No. 201410449850.X discloses a kind of composite microbial of Urban River Water pollution control
Thing preparation and preparation method thereof, the method is by by acidophilus bacillus preparation, Candida tropicalis agent and withered grass bud pole
Bacteria agent is prepared by mixing into complex micro organism fungicide, is suitable to city and asks that river COD and ammonia-nitrogen content are exceeded, river is dirty, hair
Smelly to wait the reparation of pollution and administer, it can quickly repair water ecological setting, prevent secondary pollution of water, and can quickly recover water
The self-purification function of body, clears up river bed sludge.
The patent of invention of Application No. 201510949257.6 discloses a kind of complex microorganism system for river regulation
Agent, complex microorganism includes the pure coccus of nitrous acid, thiobacillus thiooxidant, Paracoccus denitrificans, bud pole bacterium, red spirillum, lactic acid bar
Bacterium and saccharomyces cerevisiae.In complex micro organism fungicide input polluted-water, the water body smelly problem of nigrescence can be obviously improved, reduce water
Matter ammonia nitrogen and total phosphorus content.
The patent of invention of Application No. 201310126043.X discloses a kind of microbial bacteria that sewage is asked for purifying
Agent and preparation method thereof, it includes following components in parts by weight：20~40 parts of natto bud pole bacterium, 10~30 parts of nitrobacteria,
5~15 parts of white-rot fungi, 5~15 parts of pseudomonad, 10~40 parts of photosynthetic bacteria.The microbial bacterial agent has strong adaptability, energy
It is quick to form dominant microflora, supplement water body original inhabitants bacterium.COD, NH3-N, total nitrogen, TP of polluted river water etc. can be reduced in input water,
And can be deodorant except color.And noble and unsullied property biology enzyme can be produced, and suppressing the growth of harmful bacteria, accelerated decomposition residual bait, animals and plants are residual
Body, with significant effects of purification quality.
It can be seen that, continue to explore microbial bacterial agent for the research of river regulation, it is expected to alleviate or thoroughly improves increasingly serious
Water body environment pollution problem.
The content of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art, there is provided repaiied a kind of town and country river sewage original position
Multiple complex micro organism fungicide, can decompose ammonia nitrogen and nitrate in different organic matters, degradation water, reduce phosphorus element in water body
Concentration and the growth of suppression algae.
In order to solve the above technical problems, the present invention proposes following technical scheme：
A kind of complex micro organism fungicide, the complex micro organism fungicide includes pseudomonad preparation, acinetobacter calcoaceticus preparation, solution
Bacillus amyloliquefacienses preparation, Ko subtilis bar preparation, bacillus licheniformis preparation, Lactococcus lactis bacteria preparation, hydrogenlike silicon ion system
Agent, alkali lake enlightening thatch Salmonella preparation and Rhodopseudomonas palustris preparation.
Above-mentioned complex micro organism fungicide, it is preferred that pseudomonad is entrusted to be preserved in Chinese microorganism strain preservation management
Member's meeting common micro-organisms center, deposit number is the pseudomonad of CGMCC No.13433.
Above-mentioned complex micro organism fungicide, it is preferred that the parts by volume of each component is in the complex micro organism fungicide：It is false single
5~15 parts of born of the same parents' bacteria preparation, 5~15 parts of acinetobacter calcoaceticus preparation, 5~15 parts of bacillus amyloliquefaciens preparation, bacillus subtilis system
5~15 parts of agent, 5~15 parts of bacillus licheniformis preparation, 5~15 parts of Lactococcus lactis bacteria preparation, hydrogenlike silicon ion preparation 5~15
Part, 5~15 parts of alkali lake enlightening thatch Salmonella preparation, 5~15 parts of Rhodopseudomonas palustris preparation.
Above-mentioned complex micro organism fungicide, it is preferred that in the complex micro organism fungicide, total clump count >=6.0 ×
1010cfu/mL。
Above-mentioned complex micro organism fungicide, it is preferred that acinetobacter calcoaceticus are acinetobacter calcoaceticus CGMCC No.1.8587, solve starch
Bacillus is bacillus amyloliquefaciens CGMCC No.1.7463, and Bacillus subtillis is Bacillus subtillis CGMCC
No.1.2162, bacillus licheniformis are bacillus licheniformis CGMCC No.1.6510, and Lactococcus lactis are Lactococcus lactis
CGMCC No.1.3920, hydrogenlike silicon ion is hydrogenlike silicon ion CGMCC No.1.2174, and alkali lake enlightening thatch Salmonella is alkali lake enlightening thatch
Salmonella CGMCC No.1.6147, Rhodopseudomonas palustris is Rhodopseudomonas palustris CGMCC No.1.2351.
Used as a total inventive concept, the present invention also provides a kind of preparation method of above-mentioned complex micro organism fungicide,
Comprise the following steps：
(1) pseudomonad is inoculated in solid slope culture medium I and is activated, activation pseudomonad is obtained；Then will
Activation pseudomonad is inoculated in fluid nutrient medium I and is enlarged culture；It is inoculated in fermentation medium I, fermented culture system
Obtain cell concentration and be not less than 3.0 × 109The pseudomonad preparation of cfu/mL；
(2) acinetobacter calcoaceticus are inoculated in solid slope culture medium II and are activated, activation acinetobacter calcoaceticus are obtained；Then will
Activation acinetobacter calcoaceticus are inoculated in fluid nutrient medium II and are enlarged culture；It is inoculated in fermentation medium II, fermented culture
Cell concentration is obtained and is not less than 5.0 × 107The acinetobacter calcoaceticus preparation of cfu/mL；
(3) bacillus amyloliquefaciens are inoculated in solid slope culture medium III and are activated, activation solution starch bud is obtained
Spore bacillus；Then activation bacillus amyloliquefaciens are inoculated in fluid nutrient medium III and are enlarged culture；It is inoculated in fermentation training
Support in base III, fermented culture is obtained cell concentration and is not less than 8.0 × 109The bacillus amyloliquefaciens preparation of cfu/mL；
(4) Bacillus subtillis is inoculated in solid slope culture medium IV and is activated, activation Ko subtilis bar is obtained
Bacterium；Then activation Bacillus subtillis is inoculated in fluid nutrient medium IV and is enlarged culture；It is inoculated in fermentation medium IV
In, fermented culture is obtained cell concentration and is not less than 6.0 × 109The Bacillus subtillis preparation of cfu/mL；
(5) bacillus licheniformis are inoculated in solid slope culture medium V and are activated, activation lichens brood cell's bar is obtained
Bacterium；Then activation bacillus licheniformis are inoculated in fluid nutrient medium V and are enlarged culture；It is inoculated in fermentation medium V,
Fermented culture is obtained cell concentration and is not less than 8.0 × 108The bacillus licheniformis preparation of cfu/mL；
(6) Lactococcus lactis are inoculated in solid slope culture medium VI and are activated, activation Lactococcus lactis are obtained；So
Activation Lactococcus lactis are inoculated in fluid nutrient medium VI afterwards and are enlarged culture；It is inoculated in fermentation medium VI, through hair
Ferment culture is obtained cell concentration and is not less than 7.0 × 105The Lactococcus lactis bacteria preparation of cfu/mL；
(7) hydrogenlike silicon ion is inoculated in solid slope culture medium VII and is activated, activation hydrogenlike silicon ion is obtained；
Then activation hydrogenlike silicon ion is inoculated in fluid nutrient medium VII and is enlarged culture；It is inoculated in fermentation medium VII,
Fermented culture is obtained cell concentration and is not less than 8.0 × 107The hydrogenlike silicon ion preparation of cfu/mL；
(8) alkali lake enlightening thatch Salmonella is inoculated in solid slope culture medium IIX and is activated, activation alkali lake enlightening Ci Shi is obtained
Bacterium；Then activation alkali lake enlightening thatch Salmonella is inoculated in fluid nutrient medium IIX and is enlarged culture；It is inoculated in fermentation medium
In IIX, fermented culture is obtained cell concentration and is not less than 6.0 × 107The alkali lake enlightening thatch Salmonella preparation of cfu/mL；
(9) Rhodopseudomonas palustris is inoculated in solid slope culture medium IX and is activated, the activation red vacation in marsh is obtained
Monad；Then activation Rhodopseudomonas palustris is inoculated in fluid nutrient medium IX and is enlarged culture；It is inoculated in fermentation training
Support in base IX, fermented culture is obtained cell concentration and is not less than 4.0 × 107The Rhodopseudomonas palustris preparation of cfu/mL；
(10) pseudomonad preparation, acinetobacter calcoaceticus preparation, step (3) obtained in step (2) obtained in step (1) are obtained
Bacillus amyloliquefaciens preparation, Bacillus subtillis preparation, bacillus licheniformis obtained in step (5) obtained in step (4)
Preparation, Lactococcus lactis bacteria preparation, hydrogenlike silicon ion preparation, alkali obtained in step (8) obtained in step (7) obtained in step (6)
After lake enlightening thatch Salmonella preparation and Rhodopseudomonas palustris preparation obtained in step (9) proportionally mix, complex microorganism is obtained
Microbial inoculum.
The preparation method of above-mentioned complex micro organism fungicide, it is preferred that in the solid medium I, 1g/ containing peptone
100mL, dusty yeast 1.5g/100mL, glucose 2g/100mL, dipotassium hydrogen phosphate 0.1g/100mL, agar 2g/100mL, pH value 6
~8；In the solid medium II, 1.5g/100mL containing peptone, glucose 1g/100mL, sodium chloride 0.5g/100mL, fine jade
Fat 2g/100mL, pH value 6.5~7.5；In the solid medium III, 1g/100mL containing tryptone, yeast extract 0.5g/
100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6~8；In the solid medium IV, 1g/ containing tryptone
100mL, yeast extract 0.5g/100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6~8；The solid medium V
In, 1g/100mL containing peptone, yeast extract 0.5g/100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6~8；Institute
In stating solid medium VI, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/100mL, tomato juice
5g/100mL, glucose 1.0g/100mL, calcium bicarbonate 1.0g/100mL, agar 2g/100mL, pH value 4~6.5；The solid
In culture medium VII, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/100mL, agar 2g/
100mL, pH value 6.5~7.5；In the solid medium IIX, 1g/100mL containing glucose, sodium acetate 0.2g/100mL, yeast
Cream 0.5g/100mL, ammonium chloride 0.25g/100mL, agar 2g/100mL, pH value 6.5~7.5；In the solid medium IX,
1g/100mL containing glucose, sodium acetate 0.2g/100mL, yeast extract 0.5g/100mL, biotin 0.01g/100mL, magnesium sulfate
0.02g/100mL, agar 2g/100mL, pH value 6.5~7.5.
The preparation method of above-mentioned complex micro organism fungicide, it is preferred that in the fluid nutrient medium I, 1.5g/ containing glucose
100mL, yeast extract 0.5g/100mL, peptone 1g/100mL, potassium dihydrogen phosphate 0.05g/100mL, magnesium sulfate 0.04g/
100mL；In the fluid nutrient medium II, 1.5g/100mL containing glucose, yeast extract 0.5g/100mL, peptone 1g/100mL,
Sodium chloride 0.5g/100mL；In fluid nutrient medium III, 2.5g/100mL containing glucose, yeast extract 0.2g/100mL, biphosphate
Potassium 0.02g/100mL；In the fluid nutrient medium IV, 2.5g/100mL containing glucose, yeast extract 0.2g/100mL, biphosphate
Potassium 0.02g/100mL；In the fluid nutrient medium V, 2.0g/100mL containing glycerine, yeast extract 0.5g/100mL, corn pulp 1.5g/
100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium VI, 2.0g/100mL containing glucose, yeast extract 0.5g/
100mL, corn pulp 1.5g/100mL, potassium dihydrogen phosphate 0.02g/100mL, calcium bicarbonate 1.0g/100mL；The Liquid Culture
In base VII, 2.0g/100mL containing glucose, corn pulp 1.5g/100mL, ammonium sulfate 1.0g/100mL, sodium glutamate 0.2g/
100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium IIX, 1g/100mL containing glucose, sodium acetate 0.5g/
100mL, yeast extract 0.5g/100mL, ammonium chloride 0.5g/100mL, potassium nitrate 0.2g/100mL, magnesium sulfate 0.02g/100mL, phosphorus
Sour hydrogen dipotassium 0.05g/100mL；In the fluid nutrient medium IX, 1.0g/100mL containing glucose, sodium acetate 1.0g/100mL, ferment
Female cream 0.5g/100mL, ammonium chloride 0.5g/100mL, magnesium sulfate 0.05g/100mL, dipotassium hydrogen phosphate 0.02g/100mL.
The preparation method of above-mentioned complex micro organism fungicide, it is preferred that in the fermentation medium I, containing glucose 0.5~
5g/100mL, 0.5~4g/100mL of yeast extract, 1.0~6g/100mL of analysis for soybean powder, 0.01~0.1g/100mL of potassium dihydrogen phosphate,
0.01~0.1g/100mL of magnesium sulfate；In the fermentation medium II, containing 0.5~5g/100mL of glucose, yeast extract 0.3~
3g/100mL, (NH4)2SO40.1~2.0g/100mL, 0.1~1g/100mL of sodium citrate, 0.01~0.5g/ of magnesium sulfate
100mL；In the fermentation medium III, containing 0.5~5.0g/100mL of molasses, 0.3~3.0g/100mL of brown sugar, yeast extract 0.2
~2.0/100mL, 0.5~5.0/100mL of corn pulp, 0.01~0.05g/100mL of potassium dihydrogen phosphate, magnesium sulfate 0.005~
0.05g/100mL；In the fermentation medium IV, containing 0.5~5.0g/100mL of molasses, 0.5~5.0g/100mL of brown sugar, soya bean
0.5~3.0g/100mL of powder, 1.0~5.0g/100mL of corn pulp, 0.01~0.1g/100mL of potassium dihydrogen phosphate；The fermentation training
In foster base V, containing 1.0~5.0g/100mL of glycerine, 1.0~5.0/100mL of molasses, 1.0~5.0g/100mL of corn pulp, analysis for soybean powder
0.5~3.0g/100mL, 0.01~0.1g/100mL of potassium dihydrogen phosphate, 0.01~0.1g/100mL of magnesium sulfate；The fermentation training
In foster base VI, containing 1.0~5.0g/100mL of glucose, 1.0~5.0g/100mL of yeast extract, 1.0~5.0/100mL of corn pulp,
0.01~0.1g/100mL of potassium dihydrogen phosphate, 0.5~2.0g/100mL of calcium bicarbonate, 0.2~2.0g/100mL of citric acid；It is described
In fermentation medium, containing 1.0~5.0g/100mL of glucose, 1.0~5.0g/100mL of corn pulp, 1.0~5.0g/ of ammonium sulfate
100mL, 0.1~1.5g/100mL of sodium glutamate, 0.01~0.1g/100mL of potassium dihydrogen phosphate；In the fermentation medium IIX,
Containing 1.0~5.0g/100mL of glucose, 0.5~2.0g/100mL of sodium acetate, 1.0~5.0g/100mL of yeast extract, ammonium chloride 0.5
~2.0g/100mL, 0.05~0.5g/100mL of potassium nitrate, 0.01~0.1g/100mL of magnesium sulfate, dipotassium hydrogen phosphate 0.01~
0.1g/100mL, 0.1~1.0g/100mL of sodium potassium tartrate tetrahydrate；In the fermentation medium IX, containing 0.2~2.0g/ of glucose
100mL, 0.2~2.0g/100mL of sodium acetate, 0.2~2.0g/100mL of yeast extract, 0.2~2.0g/100mL of ammonium chloride, sulfuric acid
0.01~0.1g/100mL of magnesium, 0.01~0.1g/100mL of dipotassium hydrogen phosphate.
Preferably, in the step (1), the activation condition is：30~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 180rpm rotating speeds, 30 DEG C of culture 48h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~200rpm rotating speeds, 30~38 DEG C of 48~96h of culture.
Preferably, in the step (2), the activation condition is：30~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 180rpm rotating speeds, 30 DEG C of culture 48h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~160rpm rotating speeds, 30~38 DEG C of 49~120h of culture.
Preferably, in the step (3), the activation condition is：32~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 160rpm rotating speeds, 30 DEG C of culture 48h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~200rpm rotating speeds, 30~38 DEG C of 48~120h of culture.
Preferably, in the step (4), the activation condition is：32~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 150rpm rotating speeds, 37 DEG C of culture 48h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~200rpm rotating speeds, 32~38 DEG C of 48~120h of culture.
Preferably, in the step (5), the activation condition is：32~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 150rpm rotating speeds, 37 DEG C of culture 48h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~200rpm rotating speeds, 30~38 DEG C of 48~120h of culture.
Preferably, in the step (6), the activation condition is：32~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 150rpm rotating speeds, 35 DEG C of culture 72h；The fermentation culture conditions are：Rotatably shaking
Under bed 30~100rpm rotating speeds, 28~38 DEG C of 72~200h of culture.
Preferably, in the step (7), the activation condition is：32~38 DEG C, 48~72h of activation culture；The expansion
Condition of culture is：Under rotary shaker 150rpm rotating speeds, 32 DEG C of culture 72h；The fermentation culture conditions are：Rotatably shaking
Under bed 100~200rpm rotating speeds, 28~38h DEG C of 72~200h of culture.
Preferably, in the step (8), the activation condition is：32~38 DEG C, 72~96h of activation culture；The expansion
Condition of culture is：Under rotary shaker 150rpm rotating speeds, 32 DEG C of culture 72h；The fermentation culture conditions are：Rotatably shaking
Under bed 50~100rpm rotating speeds, 30~38 DEG C of 72~160h of culture.
Preferably, in the step (9), the activation condition is：32~38 DEG C, 72~96h of activation culture；The expansion
Condition of culture is：Under rotary shaker 50rpm rotating speeds, 32 DEG C of culture 96h；The fermentation culture conditions are：Rotatably shaking
Under bed 50~100rpm rotating speeds, 32~38 DEG C of 100~200h of culture.
Used as a total inventive concept, the present invention also provides a kind of above-mentioned complex micro organism fungicide or above-mentioned compound
The application of complex micro organism fungicide prepared by the preparation method of microbial bacterial agent in the river sewage in-situ immobilization of town and country.
Preferably, the application includes phosphorus element in ammonia nitrogen and nitrate, the reduction water body in decomposing organic matter, degradation water
Concentration and/or the growth of suppression algae.
Compared with prior art, the advantage of the invention is that：
1st, complex micro organism fungicide of the invention, can simultaneously decompose the ammonia nitrogen and nitre in different organic matters, degradation water
Base nitrogen, the growth for reducing phosphorus element concentration and suppression algae in water body.Town and country are carried out using complex micro organism fungicide of the invention
River sewage in-situ immobilization, with efficient, quick, non-secondary pollution, it is simple to operate the features such as, be conducive to large-scale promotion should
With.
2nd, the pseudomonad CGMCC No.13433 that complex micro organism fungicide of the invention is included are applicants by screening
A kind of efficient heterotrophic nitrification-aerobic denitrification bacterial strain for obtaining, and have efficient phenol degrading performance concurrently.Heterotrophic nitrification performance
Measurement result shows that the bacterial strain has clearance very high, and intermediate product NO3-N and NO2-N accumulation for ammonia nitrogen
Seldom；Aerobic denitrification capability measurement result shows that the bacterial strain can be fine by nitrate nitrogen and nitrite nitrogen under aerobic conditions
Removal, and intermediate product cumulant is few, non-secondary pollution.Meanwhile, SBR is applied to after bacterial strain Amplification Culture, and (batch-type is lived
Property sludge) technique, there is clearance higher to Ammonia Nitrogen in Municipal Wastewater, total nitrogen and COD, in sanitary sewage and other are each
Planting during polluted-water is administered has application value very high.Phenol degrading performance study result shows that phenol concentration is 0-
100mg/L is interval, and clearance can reach more than 95%, 100-200mg/L intervals, and it is left that a step clearance can also reach 80%
The right side, therefore, the bacterial strain has extraordinary in the water process of the exceeded simultaneous phenol pollution of ammonia nitrogen of industrial or agricultural and underground water or improvement
Application prospect.
Biomaterial preservation information
Pseudomonad (Pseudomonas sp.) of the invention, is preserved in China Microbiological bacterium on December 7th, 2016
Plant preservation administration committee's common micro-organisms center (CGMCC), address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, China
Institute of microbiology of the academy of sciences, postcode：100101.The title of the bacterial strain is：Meng Shi pseudomonad PS02, Classification And Nomenclature is
Pseudomonas monteilii PS02, deposit number is：CGMCC No.13433.
Specific embodiment
Below in conjunction with Figure of description and specific preferred embodiment, the invention will be further described, but not therefore and
Limit the scope of the invention.
Embodiment 1：Screening and performance measurement, culture and the preservation of pseudomonad CGMCC No.13433
(1) screening and performance measurement of pseudomonad CGMCC No.13433
The pseudomonad CGMCC No.13433 are from the treatment tank of Shanghai in activated sludge obtained by isolation and selection
A kind of efficient heterotrophic nitrification-aerobic denitrification bacterium, pseudomonad, preservation are accredited as through Chinese microorganism strain collection
In China Committee for Culture Collection of Microorganisms's common micro-organisms center.Specific screening process is as follows：
Activated sludge takes the treatment tank into the treatment tank of Shanghai, and 5mL muddy water mixed solutions are taken from the activated sludge
Into the heterotrophic nitrification culture medium of 45mL sterilizings.The formula of heterotrophic nitrification culture medium is：(NH4)2SO40.945g/L, citric acid
Sodium 6.536g/L, MgSO4·7H2O 1g/L, NaCl 0.12g/L, MnSO4·H2O 0.01g/L, FeSO4·7H2O 0.02g/
L, KH2PO40.2g/L, Na2HPO40.3g/L, pH 7.0~7.5.It is subsequently placed in 30 DEG C, the gas bath shaking table enrichment training of 200rpm
Support 12h.After pregnant solution is carried out into gradient dilution, heterotrophic nitrification solid medium (agar 20g/L, remaining composition are spread evenly across
With heterotrophic nitrification culture medium).After cultivating 1d in 30 DEG C of constant incubators, picking form monoclonal of different sizes is rule pure
Numbering preservation after change, more than 10 strain bacterial strains are obtained through primary dcreening operation.
By above primary dcreening operation inoculation to bromthymol blue (BTB) isolation medium.The formula of BTB culture mediums is：KNO3
1g/L, sodium succinate 8.5g/L, MgSO4·7H2O 1g/L, CaCl20.15g/L, FeSO4·7H2O 0.05g/L, KH2PO4
0.25g/L, Na2HPO40.3g/L, 1%BTB 1mL, agar 20g/L, pH 7.0~7.5.1gBTB is dissolved in 100ml absolute ethyl alcohols
Obtain final product 1%BTB ethanol solutions.After cultivating 1d in 30 DEG C of constant incubators, there is the bacterium of blue halos in picking surrounding media
Strain to BTB culture mediums, line is purified and numbering preservation.
From flat board picking colony to enriched medium, the formula of enriched medium is：Glucose 6g/L, dusty yeast 12g/L,
MgSO4·7H2O 2.5g/L, CaCl20.5g/L, KH2PO42.5g/L, pH 7.0.30 DEG C of gas bath shaking table 200rpm cultivate 12h,
1% (volume ratio) bacterium solution centrifuge washing is taken, nitrification culture medium is seeded to.Nitrify culture medium formula be：(NH4)2SO4
0.945g/L, sodium citrate 16.34g/L, MgSO4·7H2O 1g/L, KH2PO40.2g/L, Na2HPO40.3g/L.30 DEG C,
200rpm Shaking cultures, period sampling measuring ammonia nitrogen (NH4 +- N), nitrate (NO3 -- N) and nitrite (NO2 -- N) concentration,
By the heterotrophic nitrification performance for analyzing the clearance of total nitrogen (TN) to judge bacterial strain.Fig. 1 is pseudomonad CGMCC No.13433
To the removal effect figure of ammonia nitrogen, as seen from Figure 1, pseudomonad CGMCC No.13433 possess good nitrogen removal performance, 24h
When reach 98.7% to nitrogen removal rate, and nitrate nitrogen and nitrite nitrogen as intermediate product is seldom accumulated, and cumulant exists
0.02mg/L or so.
To enriched medium, 30 DEG C of gas bath shaking table 200rpm cultivate 12h to the good bacterium colony of picking Nitrification, take 1% (volume
Than) bacterium solution centrifuge washing, it is seeded to denitrification culture medium.Denitrification culture medium prescription is：KNO30.722g/L, sodium citrate
6.128g/L, MgSO4·7H2O 1g/L, KH2PO40.25g/L, Na2HPO40.3g/L.30 DEG C, 200rpm Shaking cultures are fixed
Phase is measured by sampling ammonia nitrogen (NH4 +- N), nitrate (NO3 -- N) and nitrite (NO2 -- N) concentration, by analyzing total nitrogen (TN)
Clearance judge the aerobic denitrification capability of bacterial strain.Fig. 2 is removals of the pseudomonad CGMCC No.13433 to nitrate
Design sketch, as seen from Figure 2, pseudomonad CGMCC No.13433 have good nitrogen removal performance, with nitrate as only
During one nitrogen source, its denitrification percent reaches 99.7%.
(2) cultivate
The culture medium for using for：Glucose 1.5g/100mL, yeast extract 0.5g/100mL, peptone 1g/100mL, phosphoric acid
Potassium dihydrogen 0.05g/100mL, magnesium sulfate 0.04g/100mL.Pseudomonad CGMCC No.13433 are inoculated in above-mentioned culture medium
In, under rotary shaker 150rpm rotating speeds, 32 DEG C of culture 24h obtain final product the seed for being suitable to inoculation.
(3) preserve
Added by the glycerine of high-temperature sterilization in cultured pseudomonad seed in step (2), glycerine is in seed liquor
In concentration be 30% (V/V), be placed on -80 DEG C of Refrigerator stores.
Embodiment 2
The preparation of the complex micro organism fungicide of town and country river sewage in-situ immobilization
(1) pseudomonad CGMCC No.13433 seeds are taken, is inoculated in solid medium I, activated culture, activate bar
Part is：37 DEG C, activation culture 48h is obtained activation pseudomonad CGMCC No.13433；Then will activation pseudomonad CGMCC
No.13433 is inoculated in fluid nutrient medium I and is enlarged culture, and Amplification Culture condition is：In rotary shaker 180rpm rotating speeds
Under, 30 DEG C of culture 48h；The pseudomonad seed that will be enlarged by culture is inoculated in fermentation medium I by the 10% of fermentating liquid volume,
Under rotary shaker 150rpm rotating speeds, after 32 DEG C of culture 96h, pseudomonad preparation is obtained, pseudomonad cell concentration is 4.0
×109cfu/mL。
Wherein, in solid medium I, 1g/100mL containing peptone, dusty yeast 1.5g/100mL, glucose 2g/100mL,
Dipotassium hydrogen phosphate 0.1g/100mL, agar 2g/100mL, pH value 6~8；
In fluid nutrient medium I, 1.5g/100mL containing glucose, yeast extract 0.5g/100mL, peptone 1g/100mL, phosphoric acid
Potassium dihydrogen 0.05g/100mL, magnesium sulfate 0.04g/100mL；
In fermentation medium I, 2g/100mL containing glucose, yeast extract 1.5g/100mL, analysis for soybean powder 2/100mL, di(2-ethylhexyl)phosphate
Hydrogen potassium 0.02g/100mL, magnesium sulfate 0.01g/100mL.
(2) acinetobacter calcoaceticus CGMCC No.1.8587 are taken, is inoculated in solid medium, activated culture, activation condition
For：37 DEG C, activation culture 60h is obtained activation acinetobacter calcoaceticus；Then activation acinetobacter calcoaceticus are inoculated in into fluid nutrient medium II is carried out
Amplification Culture；Amplification Culture condition is：Under rotary shaker 180rpm rotating speeds, 30 DEG C of culture 48h；Will be enlarged by the motionless of culture
Bacillus seed is inoculated in fermentation medium II by the 10% of fermentating liquid volume, under rotary shaker 160rpm rotating speeds, 32 DEG C
After culture 96h, acinetobacter calcoaceticus preparation is obtained, acinetobacter calcoaceticus cell concentration is 8.0 × 107cfu/mL。
Wherein, in solid medium II, 1.5g/100mL containing peptone, glucose 1g/100mL, sodium chloride 0.5g/
100mL, agar 2g/100mL, pH value 6.5~7.5；
In fluid nutrient medium II, 1.5g/100mL containing glucose, yeast extract 0.5g/100mL, peptone 1g/100mL, chlorine
Change sodium 0.5g/100mL；
In fermentation medium II, 3g/100mL containing glucose, yeast extract 1.5g/100mL, (NH4)2SO40.5g/100mL,
Sodium citrate 0.2g/100mL, magnesium sulfate 0.02g/100mL.
(3) bacillus amyloliquefaciens CGMCC No.1.7463 are taken, is inoculated in solid medium, activated culture, activation
Condition is：37 DEG C, activation culture 48h is obtained activation bacillus amyloliquefaciens；Then activation bacillus amyloliquefaciens are inoculated in
Fluid nutrient medium III is enlarged culture；Amplification Culture condition is：Under rotary shaker 160rpm rotating speeds, 30 DEG C of cultures
48h；The bacillus amyloliquefaciens seed that will be enlarged by culture is inoculated in fermentation medium III by the 10% of fermentating liquid volume,
Under rotary shaker 160rpm rotating speeds, after 30 DEG C of culture 72h, bacillus amyloliquefaciens preparation, bacillus amyloliquefaciens body is obtained
Concentration is 9.0 × 109cfu/mL。
Wherein, in solid medium III, 1g/100mL containing tryptone, yeast extract 0.5g/100mL, sodium chloride 1g/
100mL, agar 2g/100mL, pH value 6~8；
In fluid nutrient medium III, 2.5g/100mL containing glucose, yeast extract 0.2g/100mL, potassium dihydrogen phosphate 0.02g/
100mL；
In fermentation medium III, molasses 1.5g/100mL, brown sugar 1.0g/100mL, yeast extract 1.5/100mL, corn pulp
1.5/100mL, potassium dihydrogen phosphate 0.02g/100mL, magnesium sulfate 0.01g/100mL.
(4) Bacillus subtillis CGMCC No.1.2162 are taken, is inoculated in solid medium, activated culture activates bar
Part is：37 DEG C, activation culture 60h is obtained activation Bacillus subtillis；Then activation Bacillus subtillis is inoculated in liquid training
Support base IV and be enlarged culture；Amplification Culture condition is：Under rotary shaker 150rpm rotating speeds, 37 DEG C of culture 48h；Will be enlarged by
The Bacillus subtillis seed of culture is inoculated in fermentation medium IV by the 5% of fermentating liquid volume, in rotary shaker
Under 180rpm rotating speeds, after 37 DEG C of culture 72h, Bacillus subtillis preparation is obtained, Bacillus subtillis bulk concentration is 8.0 ×
109cfu/mL。
Wherein, in solid medium IV, 1g/100mL containing tryptone, yeast extract 0.5g/100mL, sodium chloride 1g/
100mL, agar 2g/100mL, pH value 6~8；
In fluid nutrient medium IV, 2.5g/100mL containing glucose, yeast extract 0.2g/100mL, potassium dihydrogen phosphate 0.02g/
100mL；
In fermentation medium IV, 2.0g/100mL containing molasses, brown sugar 1.5g/100mL, analysis for soybean powder 2.0g/100mL, corn
Slurry 1.5g/100mL, potassium dihydrogen phosphate 0.02g/100mL.
(5) bacillus licheniformis CGMCC No.1.6510 are taken, is inoculated in solid medium, activated culture activates bar
Part is：35 DEG C, activation culture 48h；Activation bacillus licheniformis are obtained；Then activation bacillus licheniformis are inoculated in liquid training
Support base V and be enlarged culture；Amplification Culture condition is：Under rotary shaker 150rpm rotating speeds, 37 DEG C of culture 48h；Will be enlarged by
The bacillus licheniformis seed of culture is inoculated in fermentation medium V by the 10% of fermentating liquid volume, in rotary shaker
Under 160rpm rotating speeds, after 32 DEG C of culture 96h, bacillus licheniformis preparations is obtained, bacillus licheniformis cell concentration is 1.0 ×
109cfu/mL。
Wherein, in solid medium V, 1g/100mL containing peptone, yeast extract 0.5g/100mL, sodium chloride 1g/100mL,
Agar 2g/100mL, pH value 6~8；
In fluid nutrient medium V, 2.0g/100mL containing glycerine, yeast extract 0.5g/100mL, corn pulp 1.5g/100mL, phosphoric acid
Potassium dihydrogen 0.02g/100mL；
In fermentation medium V, 2.0g/100mL containing glycerine, molasses 1.5/100mL, corn pulp 3.0g/100mL, analysis for soybean powder
1.5g/100mL, potassium dihydrogen phosphate 0.03g/100mL, magnesium sulfate 0.02g/100mL.
(6) extracting lactic acid galactococcus CGMCC No.1.3920, are inoculated in solid medium, activated culture, activation condition
For：37 DEG C, activation culture 72h is obtained activation Lactococcus lactis；Then activation Lactococcus lactis are inoculated in fluid nutrient medium VI
It is enlarged culture；Amplification Culture condition is：Under rotary shaker 150rpm rotating speeds, 35 DEG C of culture 72h；Will be enlarged by culture
Lactococcus lactis seed is inoculated in fermentation medium VI by the 10% of fermentating liquid volume, under rotary shaker 50rpm rotating speeds,
After 32 DEG C of culture 150h, Lactococcus lactis bacteria preparation is obtained, Lactococcus lactis cell concentration is 8.0 × 105cfu/mL。
Wherein, in solid medium VI, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/
100mL, tomato juice 5g/100mL, glucose 1.0g/100mL, calcium bicarbonate 1.0g/100mL, agar 2g/100mL, pH value 4~
6.5；
In fluid nutrient medium VI, 2.0g/100mL containing glucose, yeast extract 0.5g/100mL, corn pulp 1.5g/100mL,
Potassium dihydrogen phosphate 0.02g/100mL, calcium bicarbonate 1.0g/100mL；
In fermentation medium VI, 2.5g/100mL containing glucose, yeast extract 2.0g/100mL, corn pulp 3.0/100mL, phosphorus
Acid dihydride potassium 0.02g/100mL, calcium bicarbonate 1.0g/100mL, citric acid 0.5g/100mL.
(7) hydrogenlike silicon ion CGMCC No.1.2174 are taken, is inoculated in solid medium, activated culture, activation condition
For：35 DEG C, activation culture 72h；Activation hydrogenlike silicon ion is obtained；Then activation hydrogenlike silicon ion is inoculated in fluid nutrient medium
VII is enlarged culture；Amplification Culture condition is：Under rotary shaker 150rpm rotating speeds, 32 DEG C of culture 72h；Will be enlarged by training
Foster hydrogenlike silicon ion seed is inoculated in fermentation medium VII by the 10% of fermentating liquid volume, in rotary shaker 150rpm
Under rotating speed, after 32 DEG C of culture 96h, hydrogenlike silicon ion preparation is obtained, hydrogenlike silicon ion cell concentration is 9.0 × 107cfu/mL。
Wherein, in solid medium VII, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/
100mL, agar 2g/100mL, pH value 6.5~7.5；
In fluid nutrient medium VII, 2.0g/100mL containing glucose, corn pulp 1.5g/100mL, ammonium sulfate 1.0g/100mL,
Sodium glutamate 0.2g/100mL, potassium dihydrogen phosphate 0.02g/100mL；
In fermentation medium VII, 3.0g/100mL containing glucose, corn pulp 2.0g/100mL, ammonium sulfate 1.5g g/
100mL, sodium glutamate 1.5g/100mL, potassium dihydrogen phosphate 0.02g/100mL.
(8) alkali lake enlightening thatch Salmonella CGMCC No.1.6147 are taken, is inoculated in solid medium, activated culture, activate bar
Part is：32 DEG C, activation culture 96h is obtained activation alkali lake enlightening thatch Salmonella；Then activation alkali lake enlightening thatch Salmonella is inoculated in liquid training
Support base IIX and be enlarged culture；Amplification Culture condition is：Under rotary shaker 150rpm rotating speeds, 32 DEG C of culture 72h；To expand
The alkali lake enlightening thatch Salmonella seed of big culture is inoculated in fermentation medium IIX by the 15% of fermentating liquid volume, in rotary shaker
Under 50rpm rotating speeds, after 30 DEG C of culture 120h, alkali lake enlightening thatch Salmonella preparation is obtained, alkali lake enlightening thatch Salmonella cell concentration is 7.0 ×
107cfu/mL。
Wherein, in solid medium IIX, 1g/100mL containing glucose, sodium acetate 0.2g/100mL, yeast extract 0.5g/
100mL, ammonium chloride 0.25g/100mL, agar 2g/100mL, pH value 6.5~7.5；
In fluid nutrient medium IIX, 1g/100mL containing glucose, sodium acetate 0.5g/100mL, yeast extract 0.5g/100mL, chlorine
Change ammonium 0.5g/100mL, potassium nitrate 0.2g/100mL, magnesium sulfate 0.02g/100mL, dipotassium hydrogen phosphate 0.05g/100mL；
Described fermentation medium IIX components are as follows：Glucose 2.0g/100mL, sodium acetate 1.5g/100mL, yeast extract
1.0g/100mL, ammonium chloride 1.5g/100mL, potassium nitrate 0.2g/100mL, magnesium sulfate 0.03g/100mL, dipotassium hydrogen phosphate
0.02g/100mL, sodium potassium tartrate tetrahydrate 0.2g/100mL.
(9) Rhodopseudomonas palustris CGMCC No.1.2351 are taken, is inoculated in solid medium IX, activated culture is living
Change condition is：32 DEG C, activation culture 96h is obtained activation Rhodopseudomonas palustris；Then will activation Rhodopseudomonas palustris inoculation
Culture is enlarged in fluid nutrient medium IX, Amplification Culture condition is：Under rotary shaker 50rpm rotating speeds, 32 DEG C of cultures
96h；The Rhodopseudomonas palustris seed that will be enlarged by culture is inoculated in fermentation medium IX by the 15% of fermentating liquid volume, in rotation
Under rotatable shaking table 50rpm rotating speeds, after 32 DEG C of culture 160h, Rhodopseudomonas palustris preparation, Rhodopseudomonas palustris thalline is obtained
Concentration is 5.0 × 107cfu/mL。
Wherein, in solid medium IX, 1g/100mL containing glucose, sodium acetate 0.2g/100mL, yeast extract 0.5g/
100mL, biotin 0.01g/100mL, magnesium sulfate 0.02g/100mL, agar 2g/100mL, pH value 6.5~7.5；
In fluid nutrient medium IX, 1.0g/100mL containing glucose, sodium acetate 1.0g/100mL, yeast extract 0.5g/100mL,
Ammonium chloride 0.5g/100mL, magnesium sulfate 0.05g/100mL, dipotassium hydrogen phosphate 0.02g/100mL；
In fermentation medium IX, 1.5g/100mL containing glucose, sodium acetate 1.0g/100mL, yeast extract 1.5g/100mL,
Ammonium chloride 0.5g/100mL, magnesium sulfate 0.05g/100mL, dipotassium hydrogen phosphate 0.02g/100mL.
(10) by 15 parts of pseudomonad preparation, 10 parts of acinetobacter calcoaceticus preparation, step obtained in step (2) obtained in step (1)
Suddenly 8 parts of bacillus amyloliquefaciens preparation obtained in (3), 12 parts of Bacillus subtillis preparation, step (5) obtained in step (4) are made
8 parts of bacillus licheniformis preparation, 15 parts of Lactococcus lactis bacteria preparation, class ball obtained in step (7) obtained in step (6) it is red thin
12 parts of bacteria preparation, 10 parts of alkali lake enlightening thatch Salmonella preparation obtained in step (8) and Rhodopseudomonas palustris preparation obtained in step (9)
10 parts of mixing, are obtained complex micro organism fungicide, and 7.5 × 10 are approximately equal to containing viable count10cfu/mL。
Embodiment 3
Application of the complex micro organism fungicide in town and country river sewage in-situ immobilization
Suburb of Shanghai river course, river is in black, distributes strong stink, and COD, ammonia nitrogen and total phosphorus are exceeded.Taken from the river course
2L water samples, add liquid composite microbial microbial inoculum 2mL, every three days with microbial inoculum once, the Zhou Tianhou of continuous dosing three, measurement result table
Bright, water body COD drops to 15.3mg/L by delivering the 265mg/L before microorganism；Ammonia-nitrogen content is by under preprosthetic 16.8mg/L
It is down to 0.69mg/L；Total nitrogen drops to 0.97 by 31.53mg/L；Total phosphorus drops to 0.23 by 2.82mg/L, the water body after reparation
People's Republic of China's water environment quality standard V class water standards are reached.Be demonstrated by obvious water remediation effect (see
Table 1)
Repairing effect of the complex micro organism fungicide of table 1 to river sewage
| COD(mg/L) | NH3-N(mg/L) | Total nitrogen (mg/L) | Total phosphorus (mg/L) | |
| River sewage before processing | 265 | 16.8 | 31.53 | 2.82 |
| Plus after microbial bacterial agent treatment | 15.3 | 0.69 | 0.97 | 0.23 |
| Clearance | 94.22% | 95.89% | 96.23% | 91.84% |
The above is only the preferred embodiment of the present invention, and protection scope of the present invention is not limited merely to above-mentioned implementation
Example.All technical schemes belonged under thinking of the present invention belong to protection scope of the present invention.It is noted that for the art
Those of ordinary skill for, improvements and modifications under the premise without departing from the principles of the invention, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of complex micro organism fungicide, it is characterised in that the complex micro organism fungicide includes pseudomonad preparation, non-lever
Bacteria preparation, bacillus amyloliquefaciens preparation, Ko subtilis bar preparation, bacillus licheniformis preparation, Lactococcus lactis bacteria preparation, class ball
Red bacteria preparation, alkali lake enlightening thatch Salmonella preparation and Rhodopseudomonas palustris preparation.
2. complex micro organism fungicide according to claim 1, it is characterised in that pseudomonad is to be preserved in China Microbiological
Culture presevation administration committee common micro-organisms center, deposit number is the pseudomonad of CGMCC No.13433.
3. complex micro organism fungicide according to claim 1, it is characterised in that each component in the complex micro organism fungicide
Parts by volume be：5~15 parts of pseudomonad preparation, 5~15 parts of acinetobacter calcoaceticus preparation, 5~15 parts of bacillus amyloliquefaciens preparation,
5~15 parts of bacillus subtilis formulation, 5~15 parts of bacillus licheniformis preparation, 5~15 parts of Lactococcus lactis bacteria preparation, class ball are red
5~15 parts of bacteria preparation, 5~15 parts of alkali lake enlightening thatch Salmonella preparation, 5~15 parts of Rhodopseudomonas palustris preparation.
4. complex micro organism fungicide according to claim 1, it is characterised in that in the complex micro organism fungicide, total bacterium
Fall number >=6.0 × 1010cfu/mL。
5. the complex micro organism fungicide according to any one of Claims 1 to 4, it is characterised in that acinetobacter calcoaceticus are not lever
Bacterium CGMCC No.1.8587, bacillus amyloliquefaciens are bacillus amyloliquefaciens CGMCC No.1.7463, Bacillus subtillis
It is Bacillus subtillis CGMCC No.1.2162, bacillus licheniformis are bacillus licheniformis CGMCC No.1.6510, lactic acid
Galactococcus is Lactococcus lactis CGMCC No.1.3920, and hydrogenlike silicon ion is hydrogenlike silicon ion CGMCC No.1.2174, alkali lake
Enlightening thatch Salmonella is alkali lake enlightening thatch Salmonella CGMCC No.1.6147, and Rhodopseudomonas palustris is Rhodopseudomonas palustris CGMCC
No.1.2351。
6. a kind of preparation method of complex micro organism fungicide as described in any one of Claims 1 to 5, comprises the following steps：
(1) pseudomonad is inoculated in solid slope culture medium I and is activated, activation pseudomonad is obtained；Then will activation
Pseudomonad is inoculated in fluid nutrient medium I and is enlarged culture；It is inoculated in fermentation medium I, fermented culture is obtained bacterium
Bulk concentration is not less than 3.0 × 109The pseudomonad preparation of cfu/mL；
(2) acinetobacter calcoaceticus are inoculated in solid slope culture medium II and are activated, activation acinetobacter calcoaceticus are obtained；Then will activation
Acinetobacter calcoaceticus are inoculated in fluid nutrient medium II and are enlarged culture；It is inoculated in fermentation medium II, fermented culture is obtained
Cell concentration is not less than 5.0 × 107The acinetobacter calcoaceticus preparation of cfu/mL；
(3) bacillus amyloliquefaciens are inoculated in solid slope culture medium III and are activated, activation solution starch gemma bar is obtained
Bacterium；Then activation bacillus amyloliquefaciens are inoculated in fluid nutrient medium III and are enlarged culture；It is inoculated in fermentation medium
In III, fermented culture is obtained cell concentration and is not less than 8.0 × 109The bacillus amyloliquefaciens preparation of cfu/mL；
(4) Bacillus subtillis is inoculated in solid slope culture medium IV and is activated, activation Bacillus subtillis is obtained；So
Activation Bacillus subtillis is inoculated in fluid nutrient medium IV afterwards and is enlarged culture；It is inoculated in fermentation medium IV, passes through
Fermented and cultured is obtained cell concentration and is not less than 6.0 × 109The Bacillus subtillis preparation of cfu/mL；
(5) bacillus licheniformis are inoculated in solid slope culture medium V and are activated, activation bacillus licheniformis are obtained；So
Activation bacillus licheniformis are inoculated in fluid nutrient medium V afterwards and are enlarged culture；It is inoculated in fermentation medium V, through hair
Ferment culture is obtained cell concentration and is not less than 8.0 × 108The bacillus licheniformis preparation of cfu/mL；
(6) Lactococcus lactis are inoculated in solid slope culture medium VI and are activated, activation Lactococcus lactis are obtained；Then will
Activation Lactococcus lactis are inoculated in fluid nutrient medium VI and are enlarged culture；It is inoculated in fermentation medium VI, fermented training
Support prepared cell concentration and be not less than 7.0 × 105The Lactococcus lactis bacteria preparation of cfu/mL；
(7) hydrogenlike silicon ion is inoculated in solid slope culture medium VII and is activated, activation hydrogenlike silicon ion is obtained；Then
Activation hydrogenlike silicon ion is inoculated in fluid nutrient medium VII and is enlarged culture；It is inoculated in fermentation medium VII, through hair
Ferment culture is obtained cell concentration and is not less than 8.0 × 107The hydrogenlike silicon ion preparation of cfu/mL；
(8) alkali lake enlightening thatch Salmonella is inoculated in solid slope culture medium IIX and is activated, activation alkali lake enlightening thatch Salmonella is obtained；
Then activation alkali lake enlightening thatch Salmonella is inoculated in fluid nutrient medium IIX and is enlarged culture；It is inoculated in fermentation medium IIX
In, fermented culture is obtained cell concentration and is not less than 6.0 × 107The alkali lake enlightening thatch Salmonella preparation of cfu/mL；
(9) Rhodopseudomonas palustris is inoculated in solid slope culture medium IX and is activated, the activation red false unit cell in marsh is obtained
Bacterium；Then activation Rhodopseudomonas palustris is inoculated in fluid nutrient medium IX and is enlarged culture；It is inoculated in fermentation medium
In IX, fermented culture is obtained cell concentration and is not less than 4.0 × 107The Rhodopseudomonas palustris preparation of cfu/mL；
(10) by pseudomonad preparation, the obtained solution of acinetobacter calcoaceticus preparation, step (3) obtained in step (2) obtained in step (1)
Bacillus amyloliquefacienses preparation, Bacillus subtillis preparation obtained in step (4), bacillus licheniformis preparation obtained in step (5),
Lactococcus lactis bacteria preparation, hydrogenlike silicon ion preparation, alkali lake enlightening thatch obtained in step (8) obtained in step (7) obtained in step (6)
After Salmonella preparation and Rhodopseudomonas palustris preparation obtained in step (9) proportionally mix, complex micro organism fungicide is obtained.
7. the preparation method of complex micro organism fungicide according to claim 6, it is characterised in that the solid medium I
In, 1g/100mL containing peptone, dusty yeast 1.5g/100mL, glucose 2g/100mL, dipotassium hydrogen phosphate 0.1g/100mL, agar
2g/100mL, pH value 6~8；In the solid medium II, 1.5g/100mL containing peptone, glucose 1g/100mL, sodium chloride
0.5g/100mL, agar 2g/100mL, pH value 6.5~7.5；In the solid medium III, 1g/100mL containing tryptone,
Yeast extract 0.5g/100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6~8；In the solid medium IV, containing pancreas
Peptone 1g/100mL, yeast extract 0.5g/100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6~8；The solid
In culture medium V, 1g/100mL containing peptone, yeast extract 0.5g/100mL, sodium chloride 1g/100mL, agar 2g/100mL, pH value 6
~8；In the solid medium VI, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/100mL,
Tomato juice 5g/100mL, glucose 1.0g/100mL, calcium bicarbonate 1.0g/100mL, agar 2g/100mL, pH value 4~6.5；Institute
In stating solid medium VII, 0.5g/100mL containing beef extract, peptone 0.5g/100mL, yeast extract 0.5g/100mL, agar
2g/100mL, pH value 6.5~7.5；In the solid medium IIX, 1g/100mL containing glucose, sodium acetate 0.2g/100mL,
Yeast extract 0.5g/100mL, ammonium chloride 0.25g/100mL, agar 2g/100mL, pH value 6.5~7.5；The solid medium IX
In, 1g/100mL containing glucose, sodium acetate 0.2g/100mL, yeast extract 0.5g/100mL, biotin 0.01g/100mL, sulphur
Sour magnesium 0.02g/100mL, agar 2g/100mL, pH value 6.5~7.5.
8. the preparation method of complex micro organism fungicide according to claim 7, it is characterised in that the fluid nutrient medium I
In, 1.5g/100mL containing glucose, yeast extract 0.5g/100mL, peptone 1g/100mL, potassium dihydrogen phosphate 0.05g/100mL,
Magnesium sulfate 0.04g/100mL；In the fluid nutrient medium II, 1.5g/100mL containing glucose, yeast extract 0.5g/100mL, albumen
Peptone 1g/100mL, sodium chloride 0.5g/100mL；In fluid nutrient medium III, 2.5g/100mL containing glucose, yeast extract 0.2g/
100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium IV, 2.5g/100mL containing glucose, yeast extract 0.2g/
100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium V, 2.0g/100mL containing glycerine, yeast extract 0.5g/
100mL, corn pulp 1.5g/100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium VI, 2.0g/ containing glucose
100mL, yeast extract 0.5g/100mL, corn pulp 1.5g/100mL, potassium dihydrogen phosphate 0.02g/100mL, calcium bicarbonate 1.0g/
100mL；In the fluid nutrient medium VII, 2.0g/100mL containing glucose, corn pulp 1.5g/100mL, ammonium sulfate 1.0g/
100mL, sodium glutamate 0.2g/100mL, potassium dihydrogen phosphate 0.02g/100mL；In the fluid nutrient medium IIX, containing glucose
1g/100mL, sodium acetate 0.5g/100mL, yeast extract 0.5g/100mL, ammonium chloride 0.5g/100mL, potassium nitrate 0.2g/100mL,
Magnesium sulfate 0.02g/100mL, dipotassium hydrogen phosphate 0.05g/100mL；In the fluid nutrient medium IX, 1.0g/ containing glucose
100mL, sodium acetate 1.0g/100mL, yeast extract 0.5g/100mL, ammonium chloride 0.5g/100mL, magnesium sulfate 0.05g/100mL, phosphorus
Sour hydrogen dipotassium 0.02g/100mL.
9. the preparation method of the complex micro organism fungicide according to any one of claim 6~8, it is characterised in that the hair
In ferment culture medium I, containing 0.5~5g/100mL of glucose, 0.5~4g/100mL of yeast extract, 1.0~6g/100mL of analysis for soybean powder, phosphorus
Acid dihydride 0.01~0.1g/100mL of potassium, 0.01~0.1g/100mL of magnesium sulfate；In the fermentation medium II, containing glucose
0.5~5g/100mL, yeast extract 0.3~3g/100mL, (NH4)2SO40.1~2.0g/100mL, 0.1~1g/ of sodium citrate
100mL, 0.01~0.5g/100mL of magnesium sulfate；In the fermentation medium III, containing 0.5~5.0g/100mL of molasses, brown sugar
0.3~3.0g/100mL, 0.2~2.0/100mL of yeast extract, 0.5~5.0/100mL of corn pulp, potassium dihydrogen phosphate 0.01~
0.05g/100mL, 0.005~0.05g/100mL of magnesium sulfate；In the fermentation medium IV, containing 0.5~5.0g/ of molasses
100mL, 0.5~5.0g/100mL of brown sugar, 0.5~3.0g/100mL of analysis for soybean powder, 1.0~5.0g/100mL of corn pulp, di(2-ethylhexyl)phosphate
0.01~0.1g/100mL of hydrogen potassium；In the fermentation medium V, containing 1.0~5.0g/100mL of glycerine, molasses 1.0~5.0/
100mL, 1.0~5.0g/100mL of corn pulp, 0.5~3.0g/100mL of analysis for soybean powder, 0.01~0.1g/100mL of potassium dihydrogen phosphate,
0.01~0.1g/100mL of magnesium sulfate；In the fermentation medium VI, containing 1.0~5.0g/100mL of glucose, yeast extract 1.0~
5.0g/100mL, 1.0~5.0/100mL of corn pulp, 0.01~0.1g/100mL of potassium dihydrogen phosphate, 0.5~2.0g/ of calcium bicarbonate
100mL, 0.2~2.0g/100mL of citric acid；In the fermentation medium, containing 1.0~5.0g/100mL of glucose, corn pulp
1.0~5.0g/100mL, 1.0~5.0g/100mL of ammonium sulfate, 0.1~1.5g/100mL of sodium glutamate, potassium dihydrogen phosphate 0.01
~0.1g/100mL；In the fermentation medium IIX, containing 1.0~5.0g/100mL of glucose, 0.5~2.0g/ of sodium acetate
100mL, 1.0~5.0g/100mL of yeast extract, 0.5~2.0g/100mL of ammonium chloride, 0.05~0.5g/100mL of potassium nitrate, sulfuric acid
0.01~0.1g/100mL of magnesium, 0.01~0.1g/100mL of dipotassium hydrogen phosphate, 0.1~1.0g/100mL of sodium potassium tartrate tetrahydrate；It is described
In fermentation medium IX, containing 0.2~2.0g/100mL of glucose, 0.2~2.0g/100mL of sodium acetate, 0.2~2.0g/ of yeast extract
100mL, 0.2~2.0g/100mL of ammonium chloride, 0.01~0.1g/100mL of magnesium sulfate, 0.01~0.1g/ of dipotassium hydrogen phosphate
100mL。
10. as described in a kind of complex micro organism fungicide or any one of claim 6~9 as described in any one of Claims 1 to 5
The application of complex micro organism fungicide prepared by the preparation method of complex micro organism fungicide in the river sewage in-situ immobilization of town and country.
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