CN103355290A - Bacillus subtilis dry suspension agent and preparation method thereof - Google Patents
Bacillus subtilis dry suspension agent and preparation method thereof Download PDFInfo
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- CN103355290A CN103355290A CN2013103016064A CN201310301606A CN103355290A CN 103355290 A CN103355290 A CN 103355290A CN 2013103016064 A CN2013103016064 A CN 2013103016064A CN 201310301606 A CN201310301606 A CN 201310301606A CN 103355290 A CN103355290 A CN 103355290A
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Abstract
The invention relates to a bacillus subtilis dry suspension agent and a preparation method thereof. The agent comprises an effective active component, additives, and auxiliary materials. The agent is characterized in that the effective active component is bacillus subtilis T500; the additives comprise a wetting agent, a dispersion agent, an adhesion agent, and a disintegrating agent. Bacillus subtilis T500 viable cell amount in the dry suspension agent is 6.80*10<9>-8.61*10<9>cfu/g. The preparation method comprises the steps that: a bacillus subtilis T500 strain is activated, and is delivered into a seed pot and is subjected to fermentation culture; fermentation culture is carried out in a fermentation pot, such that fermentation broth is obtained; the wetting agent, the dispersion agent, the adhesion agent, and the disintegrating agent in the additives are mixed with the auxiliary agent; the mixture is well mixed by stirring with the fermentation broth according to a weight/volume ratio of 0.15:1; and the mixture is dried, such that the dry suspension agent of the bacillus subtilis is obtained.
Description
Technical field
The present invention relates to a kind of bacillus subtilis dry suspending agent and preparation method thereof.
Background technology
China is a traditional farming big country, in the crops production process, usually adopt the mode of spraying insecticide that the disease pest and weed of plant is prevented and treated, guarantee to produce good harvest, but the use of these agricultural chemicals, will cause certain harm to environment and the mankind, China's pesticide poisoning event of producing that takes place frequently also is to sound the alarm to society in recent years, and the green bactericide of developing a kind of safety, environmental protection has been instant responsibility.
Bacillus subtilis is a kind of desirable biocontrol microorganisms, and the various plants pathogen is had Antagonism, and has nontoxic, pollution-free, noresidue and be difficult for making the characteristics such as pathogen develops immunity to drugs, and has been widely used in a plurality of fields such as biopesticide.At present, develop and the bacillus subtilis formulation that puts into production have good control efficiency to multiple kinds of crops fungal diseases such as wheat sharp eyespot, rice sheath blight disease, tomato wilts.
The bacillus subtilis formulation is mainly with conventional dosage forms such as aqua, wetting powders in the market.But aqua is easy layering and precipitating in storage, is difficult for remote the popularization and preserves; Wetting powder production and to produce dust when using harmful to the operator, " caking " occur when in water, disperseing and easily stop up shower nozzle, in addition, easily produce sedimentation and cause that spray medicine front and back concentration differs in the sprinkling process, these are all seriously restricting the promotion and application of bacillus subtilis bactericide.
Dry suspending agent is a kind of safe, efficient, economic, convenient and use labour-saving novel agrochemical formulations.In the 6th international chemistry of pesticide meeting, dry suspending agent is considered to a kind of pesticides new formulation of efficient, environmental protection, is the important developing direction of formulations of pesticide processing.Compare with traditional formulations of pesticide, dry suspending agent has the advantage of solid pharmaceutical preparation and liquid preparation, and fast disintegration has preferably dispersiveness and suspension in water; The particle of its effective ingredient is thin, and the medicament of unit mass has preferably drug effect at the broad covered area of crop surface; Without dust from flying, reduce the harm to environment and operator in the production and application process, and can make the product of high concentration, reduce the storing cost, be easy to apply.
The academy of agricultural sciences, Jiangsu Province from water paddy soil, separate obtain a bacillus subtilis (
Bacillus subtilis) T-500, be preserved in the common micro-organisms center C GMCC of China Committee for Culture Collection of Microorganisms, deposit number: CGMCC NO:6058, this bacterial strain is applied for a patent on September 7th, 2012, its application number is 201210328716.5, publication number is CN102816725A, proves that through indoor flat plate mensuration and potted plant experiment this bacterial strain all has good control efficiency to rice blast, false smut and banded sclerotial blight.The pre-stage test of this bacterial strain mainly directly carries out with zymophyte bacterium liquid, therefore, relates to this bacterial strain during patent application among the embodiment, also only is confined on the using method of zymocyte liquid.At present, this bacterial strain being prepared into dry suspending agent does not appear in the newspapers.Along with country is more and more higher to the grain security demand, in order to prevent and treat the important disease of paddy rice, in the urgent need to researching and developing the new and effective viable bacteria bactericide of this bacterial strain.
Summary of the invention
The object of the invention is to: prevent and treat the needs of disease in order to satisfy paddy rice, a kind of transportation is provided and uses all very easily bacillus subtilis dry suspending agent, and the preparation method of this dry suspending agent.
The object of the present invention is achieved like this:
The dry suspending agent of a kind of bacillus subtilis, comprise effective active component, auxiliary agent and auxiliary material, it is characterized in that, described effective active composition is bacillus subtilis T500, described auxiliary agent comprises wetting agent, dispersant, binding agent, disintegrant, and bacillus subtilis T500 viable bacteria amount is 6.80 * 10 in the described dry suspending agent
9～8.61 * 10
9Cfu/g.
In the present invention, described wetting agent is fatty alcohol-polyoxyethylene ether, and described dispersant is naphthalenesulfonic acid-formaldehyde condensate, and described binding agent is sodium carboxymethylcellulose, and described disintegrant is sodium sulphate, and their mass ratio is 1:4:1:5.
In the present invention, described auxiliary material is kaolin.
The concrete steps that prepare above-mentioned dry suspending agent are as follows:
A) will cultivate after the activation of bacillus subtilis T500 bacterial strain, by inoculum concentration 1:100～1:150 cultured bacterial strain is moved into fermented and cultured 12～24h in the seeding tank, by inoculum concentration 1:100～1:150 the zymotic fluid in the seeding tank is moved into the 24～48h that ferments in the large fermentation tank again, obtain zymotic fluid;
B) wetting agent in the described auxiliary agent, dispersant, binding agent, disintegrant and auxiliary material are mixed after with step a in zymotic fluid press the weight/volume mix and blend of 0.15 ﹕ 1;
C) adopt spray drying process, with mixture in the b step at EAT 110-120 ℃, sample introduction speed 480-960ml/h, atomizing pressure 0.1-0.3MPa, hot air flow 25-35M
3Dry under the environment of/h, namely obtain the dried suspension reagent of bacillus subtilis T500.
In the present invention, EAT is 120 ℃ in the described c step, and sample introduction speed is 720ml/h, and atomizing pressure is 0.1MPa, and hot air flow is 30M
3/ h.
In the present invention, in the described b step wetting agent, dispersant, binding agent, disintegrant in mass ratio 1:4:1:5 mix with auxiliary material so that auxiliary agent accounts for 11% of amount of the mixture in the mixture.
In the present invention, described wetting agent is fatty alcohol-polyoxyethylene ether, and described dispersant is naphthalenesulfonic acid-formaldehyde condensate, and described adhesion agent is sodium carboxymethylcellulose, and described disintegrant is sodium sulphate, and described auxiliary material is kaolin.
The present invention compares with existing product and technology of preparing thereof, and the present invention has the following advantages:
1. solved existing bacillus subtilis formulation and stored the layered precipitation, dust pollution when using, the not first-class problem of concentration before and after the spray medicine.
2. divide the prepared bacillus subtilis T500 dispersive property of invention good, the viable bacteria amount is high.
3. preparation technology of the present invention compares other prilling and is easy to realize serialization production.
Description of drawings
Fig. 1 be under 54 ℃ of conditions different auxiliary agents on the impact of zymocyte liquid viable bacteria amount.
Among the figure: RT-CK is the normal temperature contrast, and 54-CK is 54 ℃ of contrasts, and A1-A5 is wetting agent, and B1-B65 is dispersant, and C1-C4 is binding agent, and D1-D6 is disintegrant.
Embodiment
Following examples are used for explanation the present invention, limit the scope of the invention but be not used in.
Bacillus subtilis T500 inoculation is cultivated after the activation of YPGA slant medium, press inoculum concentration 1:100～1:150 with fermented and cultured 12～24h in the YPG medium of cultured bacterial strain immigration seeding tank, by inoculum concentration 1:100～1:150 the zymotic fluid in the seeding tank is moved into the 24～48h that ferments in the large fermentation tank again, obtain zymotic fluid.Fermentation medium in the fermentation tank is corn starch-bean cake powder medium.
Embodiment 2
But screening used additives, the zymotic fluid that utilizes embodiment 1 to provide, adopt direct mixing method, insecticides adjuvant is mixed with fermentation of bacillus subtilis liquid with reference to applicable ratio, simultaneously take the zymocyte liquid of adding assistant not as contrast, place 14d in 54 ℃ of baking ovens, comprise 21 kinds of wetting agent, dispersant, binding agent and disintegrant four classes for the examination auxiliary agent, participate in the experiment auxiliary agent title and usage ratio see Table 1.
Table 1 screening used additives kind title and usage ratio
By as seen from Table 1, major part has decline in various degree, and wherein wetting agent A5, dispersant B 6 are larger on the impact of viable bacteria amount, descend significantly.And wetting agent A2, dispersant B 3, binding agent C3, disintegrant D2 and D4 process bacterium liquid and descend compared with the control not significantly, and it is not obvious on the bacillus subtilis impact to analyze these four kinds of auxiliary agents, and the viable bacteria amount after each auxiliary agent is cultivated as shown in Figure 1.Final selected: wetting agent is the A2 fatty alcohol-polyoxyethylene ether, and dispersant is the B3 naphthalenesulfonic acid-formaldehyde condensate, and adhesion agent is the C3 sodium carboxymethylcellulose, and described disintegrant is D4 sodium sulphate or D2 urea.
Embodiment 3
After utilizing 1 couple of bacillus subtilis T500 of embodiment bacterial strain activation culture, four kind auxiliary agent and the agricultural chemicals selected with embodiment 3 carry out mixing and stirring with filler kaolin, and acquisition viable bacteria amount is 1.8*10
9The bacterium liquid of cfu/ml.
Embodiment 4
Adopt spray drying process, by Orthogonal Optimization Test, at EAT 110-120 ℃, sample introduction speed 480-960ml/h, atomizing pressure 0.1-0.3MP, hot air flow 25-35 M
3Select optimum condition in the/h condition.
It is as shown in table 2 to record dry suspending agent viable bacteria amount and analysis result after the reaction:
Record dry suspending agent viable bacteria amount and analysis result after table 2 reaction
By as seen from Table 2, average and range analysis result can select best atomized drying condition and be: 120 ℃ of EATs, sample introduction speed 720ml/h, atomizing pressure 0.1MP, hot air flow 25M
3/ h.
Embodiment 5
Get the zymotic fluid that embodiment 1 obtains; Get wetting agent fatty alcohol-polyoxyethylene ether (peregal), dispersant naphthalenesulfonic acid-formaldehyde condensate (NNO), binding agent sodium carboxymethylcellulose (CMC), disintegrant sodium sulphate that embodiment 2 selectes, according to insecticides adjuvant ratio commonly used, mix with auxiliary material kaolin with mass ratio 1:4:1:5, so that auxiliary agent accounts for 11% of amount of the mixture in the mixture, zymotic fluid in amount of the mixture and the first step is with the weight/volume mixing and stirring of 0.15 ﹕ 1, the unit of amount of the mixture is kg, and the unit of zymotic fluid is L; Then adopt spray drying process, operating parameter is: 120 ℃ of the EATs of mixture, sample introduction speed 720ml/h, atomizing pressure 0.1MPa, hot air flow 30M
3/ h, with this understanding, present embodiment obtains in the atomized drying afterproduct, and viable count reaches 8 * 10
9Cfu/g.
Claims (7)
1. the dry suspending agent of a bacillus subtilis comprises effective active component, auxiliary agent and auxiliary material, it is characterized in that, described effective active composition be bacillus subtilis (
Bacillus subtilis) T500, described auxiliary agent comprises wetting agent, dispersant, binding agent, disintegrant, bacillus subtilis T500 viable bacteria amount is 6.80 * 10 in the described dry suspending agent
9～8.61 * 10
9Cfu/g.
2. the dry suspending agent of a kind of bacillus subtilis according to claim 1, it is characterized in that described wetting agent is fatty alcohol-polyoxyethylene ether, described dispersant is naphthalenesulfonic acid-formaldehyde condensate, described binding agent is sodium carboxymethylcellulose, described disintegrant is sodium sulphate, and their mass ratio is 1:4:1:5.
3. the dry suspending agent of a kind of bacillus subtilis according to claim 1 and 2 is characterized in that, described auxiliary material is kaolin.
4. preparation method of the dry suspending agent of bacillus subtilis as claimed in claim 1 is characterized in that:
A) will cultivate after the activation of bacillus subtilis T500 bacterial strain, by inoculum concentration 1:100～1:150 cultured bacterial strain is moved into fermented and cultured 12～24h in the seeding tank, by inoculum concentration 1:100～1:150 the zymotic fluid in the seeding tank is moved into the 24～48h that ferments in the large fermentation tank again, obtain zymotic fluid;
B) wetting agent in the described auxiliary agent, dispersant, binding agent, disintegrant and auxiliary material are mixed the weight/volume mixing and stirring that zymotic fluid is pressed 0.15 ﹕ 1 among rear and the step a;
C) adopt spray drying process, with mixture in the b step at EAT 110-120 ℃, sample introduction speed 480-960ml/h, atomizing pressure 0.1-0.3MPa, hot air flow 25-35M
3Dry under the environment of/h, namely obtain the dried suspension reagent of bacillus subtilis.
5. the preparation method of the dry suspending agent of a kind of bacillus subtilis according to claim 4 is characterized in that, EAT is 120 ℃ in the described c step, and sample introduction speed is 720ml/h, and atomizing pressure is 0.1MPa, and hot air flow is 30M
3/ h.
6. according to claim 4 or the preparation method of the dry suspending agent of 5 described a kind of bacillus subtilises, it is characterized in that, in the described b step wetting agent, dispersant, binding agent, disintegrant in mass ratio 1:4:1:5 mix with auxiliary material so that auxiliary agent accounts for 11% of amount of the mixture in the mixture.
7. the preparation method of the dry suspending agent of a kind of bacillus subtilis according to claim 6, it is characterized in that, described wetting agent is fatty alcohol-polyoxyethylene ether, described dispersant is naphthalenesulfonic acid-formaldehyde condensate, described adhesion agent is sodium carboxymethylcellulose, described disintegrant is sodium sulphate, and described auxiliary material is kaolin.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104542713A (en) * | 2014-12-22 | 2015-04-29 | 江苏省农业科学院 | Bacillus subtilis composition and application thereof |
| CN104839227A (en) * | 2015-04-08 | 2015-08-19 | 江苏省农业科学院 | Application of Bacillus amyloliquefaciens B-1619 in control of root-knot nematode disease |
| CN105076212A (en) * | 2015-09-09 | 2015-11-25 | 陕西上格之路生物科学有限公司 | Bactericidal composition containing bacillus subtilis and phenamacril and application thereof |
| CN106190920A (en) * | 2016-08-08 | 2016-12-07 | 湖南农业大学 | Bacillus subtilis YN145 and application thereof |
| CN106376599A (en) * | 2016-08-27 | 2017-02-08 | 江西天人生态股份有限公司 | Dry suspending agent of paecilomyces lilacinus and preparation method thereof |
| CN106615626A (en) * | 2017-01-10 | 2017-05-10 | 武汉新联大生物有限公司 | Probiotic dry suspension for feed and preparation method thereof |
| CN106884031A (en) * | 2017-04-25 | 2017-06-23 | 江苏省农业科学院 | To the method for the shake flask fermentation process optimization of bacillus subtilis T-500 high yield lipopeptide antibioticses |
| CN113142196A (en) * | 2020-09-04 | 2021-07-23 | 江苏省农业科学院 | Bacillus amyloliquefaciens dry suspending agent and preparation method thereof |
| CN114317329A (en) * | 2021-12-07 | 2022-04-12 | 湖北省农业科学院植保土肥研究所 | Bacillus subtilis strain and screening method and application thereof |
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Patent Citations (3)
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| CN1457643A (en) * | 2002-05-16 | 2003-11-26 | 武汉东湖高新集团股份有限公司 | Compound pesticide of biological pesticide and chemical pesticide |
| CN101643710A (en) * | 2009-08-25 | 2010-02-10 | 山东汇丰源农业科技发展有限公司 | Bacillus subtilis and application thereof |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104542713A (en) * | 2014-12-22 | 2015-04-29 | 江苏省农业科学院 | Bacillus subtilis composition and application thereof |
| CN104839227A (en) * | 2015-04-08 | 2015-08-19 | 江苏省农业科学院 | Application of Bacillus amyloliquefaciens B-1619 in control of root-knot nematode disease |
| CN105076212A (en) * | 2015-09-09 | 2015-11-25 | 陕西上格之路生物科学有限公司 | Bactericidal composition containing bacillus subtilis and phenamacril and application thereof |
| CN105076212B (en) * | 2015-09-09 | 2018-03-20 | 陕西上格之路生物科学有限公司 | A kind of bactericidal composition containing bacillus subtilis and 2-cyano-3-amino-3-phenylancryic acetate |
| CN106190920A (en) * | 2016-08-08 | 2016-12-07 | 湖南农业大学 | Bacillus subtilis YN145 and application thereof |
| CN106190920B (en) * | 2016-08-08 | 2019-08-27 | 湖南农业大学 | Bacillus subtilis YN145 and its application |
| CN106376599A (en) * | 2016-08-27 | 2017-02-08 | 江西天人生态股份有限公司 | Dry suspending agent of paecilomyces lilacinus and preparation method thereof |
| CN106615626A (en) * | 2017-01-10 | 2017-05-10 | 武汉新联大生物有限公司 | Probiotic dry suspension for feed and preparation method thereof |
| CN106884031A (en) * | 2017-04-25 | 2017-06-23 | 江苏省农业科学院 | To the method for the shake flask fermentation process optimization of bacillus subtilis T-500 high yield lipopeptide antibioticses |
| CN106884031B (en) * | 2017-04-25 | 2020-12-11 | 江苏省农业科学院 | Method for optimizing shake flask fermentation process of bacillus subtilis T-500 for high yield of lipopeptide antibiotics |
| CN113142196A (en) * | 2020-09-04 | 2021-07-23 | 江苏省农业科学院 | Bacillus amyloliquefaciens dry suspending agent and preparation method thereof |
| CN114317329A (en) * | 2021-12-07 | 2022-04-12 | 湖北省农业科学院植保土肥研究所 | Bacillus subtilis strain and screening method and application thereof |
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