CN115316404A - Dispersible oil suspending agent containing pyrethrin and beauveria bassiana and application thereof - Google Patents
Dispersible oil suspending agent containing pyrethrin and beauveria bassiana and application thereof Download PDFInfo
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Abstract
The invention relates to a dispersible oil suspending agent containing pyrethrin and beauveria bassiana and application thereof, belonging to the technical field of biological pesticide and biological pest control. The dispersible oil suspending agent of the pyrethrin and the beauveria bassiana comprises the following components in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 10 to 13 percent of emulsifier, 7 to 8 percent of aqueous dispersant, 3 to 5 percent of non-aqueous dispersant, 3.5 to 5 percent of rheological modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base. The dispersible oil suspending agent is prepared from natural substances, is environment-friendly, has excellent and stable physical and chemical properties, has a good homoptera control effect, especially has an optimal control effect on aphids, is low in active ingredient consumption, and can remarkably reduce the production cost.
Description
Technical Field
The invention relates to a dispersible oil suspending agent containing pyrethrin and beauveria bassiana and application thereof, belonging to the technical field of biological pesticides and biological pest control.
Background
The pesticide plays a key role in pest control and is an important guarantee for agricultural production. With the progress of science and technology, people have stronger environmental protection and health consciousness, and natural and low-toxicity pesticides are gradually replacing high-toxicity pesticides. Entomogenous fungi is a natural biological pesticide with low toxicity and can effectively control harmful insects, such as Beauveria bassiana (Beauveria bassiana), has strong adaptability and wide host range, can infect thousands of insects and mites, but has slow and unstable insecticidal effect.
The natural pyrethrin is a plant secondary compound with insecticidal activity separated from pyrethrum flower, and comprises six effective components of pyrethrin I and II, guayulin I and II, and allethrin I and II. The natural pyrethrin has the advantages of broad insecticidal spectrum, rapidity, environmental friendliness and the like, but is easy to photolyze, short in lasting period and expensive in price, and the development and application of the natural pyrethrin are limited.
The natural pyrethrin and the beauveria bassiana are mixed, so that the characteristic of quick pest killing by the pyrethrin is exerted, and the defect of slow response of the beauveria bassiana to pests is overcome; meanwhile, the dosage of the natural pyrethrin is reduced, the cost is reduced, the trouble of tank mixing of pesticide fields is avoided, and the time and the cost are saved. Therefore, the development of the mixed preparation of the natural pyrethrin and the beauveria bassiana has wide development prospect.
The application of the active ingredients of the pesticide can be used in the field only by processing the active ingredients into preparations, and the mixed composition of the entomogenous fungi and the chemical pesticide in the prior art is processed into the preparation, so that various defects still exist, for example, the activity of the entomogenous fungi is influenced by an auxiliary agent in the preparation, so that the insecticidal effect is low; for example, chemical pesticides are not highly soluble or insoluble in adjuvants, and adjuvants affect the emulsifying dispersibility and stability of active ingredients of pesticides, thereby affecting the performance of preparations and further the insecticidal effect of the preparations.
In conclusion, the research and development of the pesticide preparation prepared by mixing the natural pyrethrin and the beauveria bassiana with stable performance and good effect has important significance.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana, and the dispersible oil suspending agent has stable performance, good suspension performance and obvious aphid prevention and treatment effect.
In order to realize the purpose, the invention adopts the technical scheme that: the dispersible oil suspending agent comprises the following components in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 8.5 to 14.5 percent of emulsifier, 7 to 8 percent of aqueous dispersant, 3 to 5 percent of non-aqueous dispersant, 3.5 to 5 percent of rheology modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
Preferably, the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 10 to 12 percent of emulsifier, 7.5 percent of water-based dispersant, 4 percent of non-water-based dispersant, 4 to 5 percent of rheology modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
Preferably, the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 percent of natural pyrethrin, 0.7 percent of beauveria bassiana conidia, 11 percent of emulsifier, 7.5 percent of aqueous dispersant, 4 percent of non-aqueous dispersant, 4 percent of rheology modifier, 3 percent of synergist, 0.007 percent of ultraviolet protective agent and 69.493 percent of oil base.
Preferably, the emulsifier is a nonionic surfactant.
Preferably, the aqueous dispersant includes at least one of a polyoxyethylene ether-based dispersant and polyoxyethylene (40) sorbitol hexaoleate.
Preferably, the non-aqueous dispersant is at least one of an alkoxyethylene (5) propylene (15) based sorbitol ester, a polyalkylene block copolymer and a star structure copolymer.
Preferably, the rheology modifier comprises at least one of diatomaceous earth, xanthan gum, sodium carboxymethylcellulose, organobentonite, polyester block copolymer and fumed silica.
Preferably, the synergist is a synergist amine.
Preferably, the UV protective agent comprises zinc oxide, 2- (3, 5-di-tert-amyl-2-hydroxyphenyl) benzotriazole and titanium dioxide (TiO) 2 ) At least one of (a).
Preferably, the oil base comprises at least one of sunflower oil, soybean oil, peanut oil, and a vegetable blend oil.
Alternatively, the nonionic surfactant includes at least one of polyoxyethylene (40) sorbitol hexaoleate, polyalkylene block copolymers, castor oil polyoxyethylene ether castor oil polyoxyethylene (10) ether, castor oil polyoxyethylene ether castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene (30) ether, castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene ether, alkoxy ethylene (5) propylene (15) sorbitan ester, polyalkylene block copolymers, star-structured copolymers, polyethylene glycol 400 monooleate, and polyoxyethylene (40) perennial sorbitol ester.
Alternatively, the nonionic surfactants include castor oil polyoxyethylene (40) ether, polyethylene glycol 400 monooleate, and polyoxyethylene (40) perennial sorbitol ester; the mass ratio of the castor oil polyoxyethylene (40) ether to the polyethylene glycol 400 monooleate to the polyoxyethylene (40) perennial sorbitol ester is (7-8): (2-3): (1-2).
Alternatively, the polyoxyethylene ether-based dispersant is a polyalkylene block copolymer.
Optionally, the aqueous dispersant comprises polyoxyethylene (40) sorbitol hexaoleate
The non-aqueous dispersant comprises a star-structure copolymer, an alkoxyethylene (5) propylene (15) sorbitol ester and the non-ionic block copolymer, wherein the mass ratio of the star-structure copolymer, the alkoxyethylene (5) propylene (15) sorbitol ester to the non-ionic block copolymer is (3-5): (0-1): (0-2).
Optionally, the rheology modifier comprises an organobentonite clay, a polyester block copolymer, and fumed silica; the mass ratio of the organic bentonite to the polyester block copolymer to the fumed silica is (2-3): 1:1.
Optionally, the oil base is a vegetable blend oil comprising soybean oil, peanut oil, and sesame oil; the mass ratio of the soybean oil to the peanut oil to the sesame oil is 90.
The invention also aims to provide the application of the dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana in preventing and controlling agricultural pests, wherein the agricultural pests are homoptera pests, and particularly have obvious effect on preventing and controlling aphids.
Compared with the prior art, the invention has the beneficial effects that:
(1) The dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana provided by the invention has stable performance and good suspension performance; the auxiliary agent in the dispersible oil suspending agent has no obvious influence on the safety of the effective component beauveria bassiana and can be mutually dissolved with the pyrethrin, so that the insecticidal effect of the combination of the pyrethrin and the beauveria bassiana is not influenced;
(2) The dispersible oil suspending agent containing pyrethrin and beauveria bassiana provided by the invention has a remarkable aphid prevention and treatment effect;
(3) The dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana provided by the invention has the advantages of natural components, environmental friendliness, excellent and stable physical and chemical properties, good aphid control effect, low effective component consumption and capability of obviously reducing the production cost.
Drawings
FIG. 1 is the solubility of pyrethrin in an oil base; wherein the oil base in the test tube 1 is sunflower seed oil, the oil base in the test tube 2 is soybean oil, the oil base in the test tube 3 is peanut oil, the oil base in the test tube 4 is ethyl oleate, the oil base in the test tube 5 is methyl oleate, and the oil base in the test tube 6 is vegetable blend oil;
FIG. 2 is the appearance of four dispersible oil suspensions of examples 6 and 7 and comparative examples 1 and 2 after hot storage; wherein fig. 2A is the dispersible oil suspension prepared in example 6, fig. 2B is the dispersible oil suspension prepared in example 7, fig. 2C is the dispersible oil suspension prepared in comparative example 1, and fig. 2D is the dispersible oil suspension prepared in comparative example 2.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to the following detailed drawings and examples. In the examples, the experimental methods used were all conventional methods unless otherwise specified, and the materials, reagents and the like used were commercially available without otherwise specified.
The beauveria bassiana conidium powder of the invention comprises the following components: rice fermentation product from BbPs01 strain (Guangdong province microorganism culture collection accession number GDMCC 61494), and has spore content of (1.77 + -0.31) × 10 11 The spore germination rate is (93.75 +/-1.92)%, the drying decrement is (2.53 +/-0.43)%, and other indexes meet the requirements, so the method can be used for processing dispersible oil suspending agents.
The pyrethrin crude oil of the invention has the natural pyrethrin content of 71.78 percent and is purchased from Yunnan treasure Biotechnology Limited liability company.
The vegetable blend oil provided by the invention is vegetable blend oil and comprises 90% of soybean oil, 9% of peanut oil and 1% of sesame oil by mass percentage.
The embodiment of the invention provides a dispersible oil suspending agent containing pyrethrin and beauveria bassiana, and the dispersible oil suspending agent comprises the following raw materials in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 8.5 to 14.5 percent of emulsifier, 7 to 8 percent of aqueous dispersant, 3 to 5 percent of non-aqueous dispersant, 3.5 to 5 percent of rheological modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
In one embodiment, the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 10 to 12 percent of emulsifier, 7.5 percent of water-based dispersant, 4 percent of non-water-based dispersant, 4 to 5 percent of rheology modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
More specifically, the dispersible oil suspending agent comprises the following raw materials in percentage by mass: 0.3 percent of natural pyrethrin, 0.7 percent of beauveria bassiana conidia, 11 percent of emulsifier, 7.5 percent of aqueous dispersant, 4 percent of non-aqueous dispersant, 4 percent of rheology modifier, 3 percent of synergist, 0.007 percent of ultraviolet protective agent and 69.493 percent of oil base.
In one embodiment, the emulsifier is a nonionic surfactant.
In one embodiment, the aqueous dispersant includes at least one of a polyoxyethylene ether-based dispersant and polyoxyethylene (40) sorbitol hexaoleate.
In one embodiment, the non-aqueous dispersant is at least one of an alkoxyethylene (5) propylene (15) based sorbitol ester, a polyalkylene block copolymer, and a star-structured copolymer.
In one embodiment, the rheology modifier comprises at least one of diatomaceous earth, xanthan gum, sodium carboxymethylcellulose, organobentonite, polyester block copolymer, and fumed silica.
In one embodiment, the synergist is a synergist amine.
In one embodiment, the UV protectant includes zinc oxide, 2- (3, 5-di-tert-amyl-2-hydroxyphenyl) benzotriazole, and titanium dioxide (TiO) 2 ) At least one of (1).
In one embodiment, the oil base comprises at least one of sunflower oil, soybean oil, peanut oil, and a vegetable blend oil.
In one embodiment, the nonionic surfactant comprises at least one of polyoxyethylene (40) sorbitan hexaoleate, polyalkylene block copolymers, castor oil polyoxyethylene ether castor oil polyoxyethylene (10) ether, castor oil polyoxyethylene ether castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene (30) ether, castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene ether, an alkoxy ethylene (5) propylene (15) sorbitan ester, polyalkylene block copolymers, star copolymers, polyethylene glycol 400 monooleate, and polyoxyethylene (40) perennial sorbitan ester.
More specifically, the nonionic surfactants include castor oil polyoxyethylene (40) ether, polyethylene glycol 400 monooleate, and polyoxyethylene (40) perennial sorbitol ester; the mass ratio of the castor oil polyoxyethylene (40) ether to the polyethylene glycol 400 monooleate to the polyoxyethylene (40) perennial sorbitol ester is (7-8): (2-3): (1-2).
In one embodiment, the polyoxyethylene ether-based dispersant is a polyalkylene block copolymer.
In one embodiment, the aqueous dispersant comprises polyoxyethylene (40) sorbitol hexaoleate.
More specifically, the non-aqueous dispersant includes a star-structured copolymer; or the non-aqueous dispersant comprises a star-structure copolymer, an alkoxyethylene (5) propylene (15) sorbitol ester and the non-ionic block copolymer, wherein the mass ratio of the star-structure copolymer, the alkoxyethylene (5) propylene (15) sorbitol ester and the non-ionic block copolymer is (3-5): (0-1): (0-2).
More specifically, the rheology modifiers include organobentonites, polyester block copolymers, and fumed silica; the mass ratio of the organic bentonite to the polyester block copolymer to the fumed silica is (2-3): 1:1.
More specifically, the oil base is a vegetable blend oil comprising soybean oil, peanut oil and sesame oil; the mass ratio of the soybean oil to the peanut oil to the sesame oil is 90.
The embodiment of the invention provides application of a dispersible oil suspending agent containing pyrethrin and beauveria bassiana in prevention and control of agricultural pests, wherein the agricultural pests are homoptera pests, and particularly have an obvious effect on preventing and controlling aphids.
In order to explain the technical solution of the present invention more clearly and in detail, the following examples are provided for further explanation.
Example 1: aiming at the dispersible oil suspending agent containing pyrethrin and beauveria bassiana of the invention, the screening of the assistant oil base
(1) Safety to Beauveria bassiana
Taking 20mL of each test oil base (soybean oil, peanut oil, sunflower oil, methyl oleate, ethyl oleate, castor oil, rapeseed oil and vegetable blend oil) into a clean 50mL centrifuge tube, drying in vacuum, sealing and sterilizing. Adding 1g of spore powder into each centrifuge tube respectively, mixing uniformly, suspending, storing at normal temperature, taking no treatment as a blank control, repeating each treatment for 3 times, and determining the spore germination rate, wherein the results are shown in table 1.
TABLE 1 Effect of test oil base on spore germination of Beauveria bassiana
Note: the data in the table are mean ± sd, and different letters after the same column indicate significant differences at P < 0.05 level (new repolarization test, DMRT).
Test results show that in 8 oil bases to be tested, compared with the oil bases stored for 7 days, after the oil bases are stored for 30 days, the germination rates of the beauveria bassiana spores are remarkably reduced to 23.31%, 5.15%, 59.49% and 57.14% respectively; soybean oil, peanut oil, sunflower seed oil and plant blend oil have no influence on the vitality of beauveria bassiana spores, and can be used as an oil base of a dispersible oil suspending agent containing pyrethrin and beauveria bassiana.
(2) Solubility to pyrethrin
On the basis of the oil-based screening experiment, sunflower seed oil, soybean oil, peanut oil, ethyl oleate, methyl oleate and vegetable blend oil are respectively mixed with pyrethrin crude oil in a test tube, and whether the layering phenomenon exists or not is observed, and the result is shown in figure 1.
As shown in figure 1, the test results show that all the tested oil bases (sunflower oil, soybean oil, peanut oil, ethyl oleate, methyl oleate, vegetable blend oil) can be mutually dissolved with the pyrethrin crude oil without layering,
in conclusion, the sunflower seed oil, the soybean oil, the peanut oil and the plant blend oil can be used as the oil base of the dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana.
Example 2: aiming at the dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana, the invention screens the rheological modifier
(1) Safety assay for Beauveria bassiana
Taking a culture dish with filter paper with a proper size paved at the bottom, weighing 500mg of spore powder, placing the spore powder on the filter paper, weighing 500mg of various rheological modifiers (diatomite, xanthan gum, sodium hydroxymethyl cellulose, organic bentonite, polyester block copolymer and fumed silica), respectively and uniformly mixing the various rheological modifiers with the spore powder, taking the spore powder without the rheological modifiers as a reference, covering a sealing film for sealing and storing, repeating the steps for 3 times, standing for 7d and 15d at normal temperature, and then measuring the spore germination rate, wherein the test results are shown in table 2.
TABLE 2 Effect of test rheology modifiers on spore viability of Beauveria bassiana
Note: the data in the table are mean ± sd, and different letters after the same column indicate significant differences at P < 0.05 level (new repolarization test, DMRT).
As can be seen from Table 2, the test results show that the compatibility of various tested rheology modifiers with beauveria bassiana spores is good, the germination rates of the beauveria bassiana spores in the treatment and the control groups have no significant difference, and the germination rates of the beauveria bassiana spores are all over 70%.
However, considering that the diatomite has a special porous adsorption effect, it has a certain suspension property; xanthan gum and sodium hydroxymethyl cellulose are dissolved in water to show the thickening property; therefore, the rheology modifier of dispersible oil suspension containing pyrethrin and beauveria bassiana is preferably selected from organobentonite, polyester block copolymer and fumed silica.
(2) Optimization of rheology modifiers
The rheology modifier was optimized by orthogonal test, 3 factors were set, 3 levels were set, 9 treatments were performed, and the amounts of organobentonite, polyester block copolymer and fumed silica were as shown in table 3, in each treatment, 2% of conidia powder of beauveria bassiana, 5% or 2.5% of a non-aqueous dispersant (star-structured copolymer) and the balance of oil base (vegetable blend oil) were added in mass%. The experiment was repeated 3 times. The oil separation rate, pourability and suspension rate were investigated according to the relevant standards.
Table 3 rheology modifier optimized orthogonal design table
TABLE 4 rheology modifier optimization test results (when the star-structured copolymer contains 5%)
Note: the data in the table are mean ± sd, same level and different letters after same column data indicate significant difference at P < 0.05 level (new repolarization test, DMRT).
TABLE 5 rheology modifier optimization test results (when the star copolymer contains 2.5%)
Note: the data in the table are mean ± sd, same level and different letters after same column data indicate significant difference at P < 0.05 level (new repolarization test, DMRT).
The experimental results are shown in tables 4 and 5, the organic bentonite has the largest influence on the oil separation rate, the range of the organic bentonite is obviously higher than that of the other two rheology modifiers, and the optimal dosage of the organic bentonite is 2-3%; the polyester block copolymer has the greatest effect on the suspension percentage, and the amount thereof is optimally 1%. The fumed silica has a large influence on the oil separating rate and small influence on the pourability and the suspension rate, and the dosage of the fumed silica is preferably 0.5 to 1 percent.
Therefore, the rheology modifier of the dispersible oil suspending agent containing pyrethrin and beauveria bassiana is selected from organic bentonite, polyester block copolymer and fumed silica; and the mass ratio of the organic bentonite to the polyester block copolymer to the fumed silica is (2-3) to 1 (0.5-1).
Example 3: aiming at the dispersible oil suspending agent containing pyrethrin and beauveria bassiana of the invention, the screening of an emulsifier and a dispersant is carried out
(1) Evaluation of emulsion dispersibility
A mixture of vegetable blend oils containing 15% surfactant was prepared by placing 40mL of vegetable blend oil in 50mL centrifuge tubes, adding 6mL of each surfactant to be tested (as shown in Table 6), and observing the dispersion during the preparation. Taking 1mL of the mixture in a centrifuge tube filled with 39mL of water, shaking up, observing the emulsification condition, standing for 30min, and recording and observing the emulsification dispersion stability of the sample.
Evaluation criteria for emulsion dispersibility: first-stage, quickly and automatically uniformly dispersing, and slightly stirring to obtain fluorescent blue or light white transparent emulsion; and (2) second stage: can automatically and uniformly disperse, and is in a blue or light milky semitransparent emulsion by slight stirring; third stage: white cloud or filiform dispersion, and slightly stirring to obtain light blue or milky opaque emulsion; four stages: the white particles sink and become opaque emulsion after stirring; and (5) fifth stage: sink in oil form, emulsify incompletely with stirring, and separate layers quickly after stopping.
The results are shown in table 6, except that fatty alcohol polyoxyethylene ether (N = 9), alkoxy phosphate and nonylphenol polyoxyethylene ether NP-10 are poor and have more than four-order dispersibility in oil or water, and the remaining surfactants tested have good dispersibility in both oil and water.
TABLE 6 evaluation of Dispersion emulsifiability of test surfactant
(2) Safety assay for Beauveria bassiana
200 mu L of various emulsifiers to be tested are respectively dripped into 50mL centrifuge tubes filled with 20mL of distilled water to prepare aqueous solutions, the distilled water without a surfactant is used as a control group, 30mg of beauveria bassiana conidium powder is respectively added under aseptic conditions, a high-speed oscillator is used for uniformly oscillating, the conidium powder is stored at normal temperature of 1d,7d and 15d, then the spore germination rate is measured, and each treatment is repeated for 3 times. The results are shown in Table 7, and the surfactants tested have no influence on the spore germination activity of beauveria bassiana except for nonylphenol polyoxyethylene ether NP-10, nonylphenol polyoxyethylene ether NP-7, fatty alcohol polyoxyethylene ether (N = 9) and octylphenol polyoxyethylene ether OP-4.
TABLE 7 influence of test surfactants on the germination capacity of conidia of Beauveria bassiana
Note: the data in the table are mean ± sd, and different letters after the same column indicate significant differences at P < 0.05 level (new repolarization test, DMRT).
The results in tables 6 and 7 show that the surfactants such as polyoxyethylene (40) sorbitol hexaoleate, nonionic block copolymer, castor oil polyoxyethylene (10) ether, castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene (30) ether, castor oil polyoxyethylene ether, alkoxy ethylene (5) propylene (15) sorbitol ester, polyalkylene block copolymer, star structure copolymer, polyethylene glycol 400 monooleate and polyoxyethylene (40) perennial sorbitol ester among the nonionic surfactants have no significant influence on the germination force of beauveria bassiana conidia and the evaluation of emulsifying dispersibility is good.
In the experimental process, the beauveria bassiana spores emulsified by the castor oil polyoxyethylene (40) ether have a faster germination speed than the castor oil polyoxyethylene (30) ether; the castor oil polyoxyethylene ether is easy to solidify at low temperature, and is not beneficial to subsequent production and processing. Therefore, castor oil polyoxyethylene (30) ether and castor oil polyoxyethylene ether are removed, and other surfactants are used for further screening the emulsifying agent and the dispersing agent.
(3) Screening of emulsifier and non-aqueous dispersant
The mass fraction of the effective components is 1% (0.7% beauveria bassiana conidium powder and 0.3% natural pyrethrin), and the rheology modifier is polyester block copolymer, fumed silica and organic bentonite composition, which respectively account for 1%, 1% and 2%. 5 levels of 6 factor orthogonal tests were set, as in Table 8, and the experiments were repeated 3 times, and the oil recovery and suspension rate for each treatment were determined following the relevant standards.
Experimental results it was found that the emulsifiers tested and the non-aqueous dispersants had different degrees of influence on the oil separation and suspension ratios as shown in tables 9 and 10:
when the content of the castor oil polyoxyethylene (40) ether is less than 5%, the emulsifying effect is poor, white insoluble droplets are formed after the castor oil polyoxyethylene (40) ether is dissolved in water, the oil separation rate is high, the suspension rate is low, and the content effect of 5-10% is good; the dosage of the polyoxyethylene 400 monooleate has great influence on oil separation and suspension property, and the effect is best when the content is 2.5 percent; the polyoxyethylene (40) perennial acid sorbitol ester content is 1% -2%.
Therefore, the emulsifier of the pyrethrin and beauveria bassiana dispersible oil suspending agent is castor oil polyoxyethylene (40) ether, polyethylene glycol 400 monooleate and polyoxyethylene (40) sorbitan monooleate; the mass ratio of the castor oil polyoxyethylene (40) ether to the polyethylene glycol 400 monooleate to the polyoxyethylene (40) perennial sorbitol ester is (7-8) to (2-3) to (1-2).
Non-aqueous dispersant: the alkoxy ethylene propylene sorbitol ester has certain influence on the oil separating rate, but has no influence on the suspension rate, and the dosage can be considered to be less than 1%; the nonionic block copolymer and the star-structured copolymer can control the oil separating problem with the increase of the dosage, but the suspension percentage is reduced, the system becomes more viscous and is not easy to pour, and in contrast, the star-structured copolymer has stronger oil separating control capability than the nonionic block copolymer, so that the dosage of the star-structured copolymer is 3-5 percent, and the dosage of the nonionic block copolymer is 0-2 percent.
Therefore, the non-aqueous dispersant of the dispersible oil suspending agent containing pyrethrin and beauveria bassiana comprises a star-structure copolymer, alkoxy ethylene propylene sorbitol ester and a non-ionic block copolymer, wherein the mass ratio of the star-structure copolymer, the alkoxy ethylene propylene sorbitol ester and the non-ionic block copolymer is (3-5) to (0-1) to (0-2).
TABLE 8 emulsifier and non-aqueous dispersant optimization test
TABLE 9 (%) effect of surfactant on oil deposition
Note: different letters after the same column of data in the table indicate significant differences at the level of P < 0.05.
TABLE 10 Effect of surfactants on suspension Rate (%)
Note: the table shows significant differences at P < 0.05 levels with different letters after the same column of data (New Bipolar Difference test, DMRT).
(4) Aqueous dispersant screening
The mass fraction of the effective components is 1% (0.7% beauveria bassiana conidium powder and 0.3% natural pyrethrin), the rheology modifier is polyester block copolymer, fumed silica and organic bentonite composition, and the compositions respectively account for 1%, 1% and 2%; 7.5% of castor oil polyoxyethylene (40) ether as an emulsifier, 2.5% of polyethylene glycol 400 monooleate and 1% of polyoxyethylene (40) perennial sorbitol ester; the dosage of the oily dispersant is 4 percent of the dosage of the star-shaped structure copolymer. Two aqueous dispersants, polyalkylene block copolymer and polyoxyethylene sorbitol hexaoleate, were selected for testing, and the experiment was repeated 3 times to compare their effects on oil separation, suspension and pour residue.
The results show that the polyalkylene block copolymer significantly affects the pouring residue amount when the amount is more than 5% as shown in table 11, while the polyoxyethylene sorbitol hexaoleate is excellent in combination, and the amount is most preferably 7.5%.
Therefore, the water-based dispersant of the dispersible oil suspending agent with the pyrethrin and the beauveria bassiana is polyoxyethylene sorbitol hexaoleate, and the using amount of the water-based dispersant is 7-8%.
TABLE 11 comparison of the Effect of aqueous dispersants
* T test results are shown, indicating that the mean is significantly higher than another mean in the same column.
Example 4: aiming at the dispersible oil suspending agent containing pyrethrin and beauveria bassiana of the invention, the ultraviolet protective agent is screened
Preparing 1% pyrethrin (0.3%) and beauveria bassiana (0.7%) dispersible oil suspending agents according to the screened oil base, emulsifier, dispersant and rheological modifier and the dosage thereof, respectively adding different ultraviolet protective agents to be tested to the set dosage, storing for 15 days at normal temperature, diluting the preparation, performing ultraviolet treatment (irradiating a position 30cm below a 30W ultraviolet lamp for a certain time), measuring the spore germination rate at intervals, taking the ultraviolet protective agent as a control group (CK) without adding, and repeating the experiment for 3 times.
The experimental results show that, as shown in table 12, if there is no ultraviolet treatment, other test substances, except the fluorescent whitening agent 33, have no effect on spore germination rate; if the ultraviolet treatment is given for 0.5 to 2 hours, various test substances have a significant protective effect on spore germination, especially titanium dioxide (TiO) 2 ) The effect is best, and the dosage is preferably 0.005 to 0.01 percent.
TABLE 12 influence of UV-protective agents on the germination rate (%) of Beauveria bassiana spores
Note: the data in the table are mean ± standard deviation, with non-identical letters after the same column of data indicating significant differences at P < 0.05 (new repolarization test, DMRT).
Example 5: aiming at the dispersible oil suspending agent containing the pyrethrin and the beauveria bassiana, the invention screens the synergist
(1) Determination of safety to Beauveria bassiana
According to the various auxiliary agents and the dosage screened by the test, 1 percent of pyrethrin (0.3 percent) and 0.7 percent of beauveria bassiana dispersible oil suspending agent are prepared, and respectively added with a set dosage of synergistic amine 1 (Zheng Zhou sea chemical engineering) or synergistic amine 2 (Wuhan Huake Jiejie biotechnology), a sealing film is sealed, stored at normal temperature and repeated for 3 times, and no synergistic agent is added as a control group (CK). The spore germination rate was measured at intervals and the results are shown in Table 13.
TABLE 13 influence of Amines on spore germination rates of Beauveria bassiana
Note: the data in the table are mean ± standard deviation, with the same letter after the same column data indicating no significant difference at P < 0.05 level (new repolarization test, DMRT).
(2) Determination of the Effect
The liquid medicine prepared by the method is used, biological measurement is carried out on the Aphis gossypii Glover by respectively adopting a spraying method to dilute 1000 times, the preparation without the synergist and clean water are used as a control group, the biological measurement is repeated for 3 times, the death condition of the Aphis gossypii Glover is observed and recorded every 24 hours, and the corrected death rate of the Aphis gossypii Glover is calculated, and the results are shown in table 14.
TABLE 14 Effect test of potentiators
Note: the data in the table are mean ± sd, and different letters after the same column indicate significant differences at P < 0.05 level (new repolarization test, DMRT).
Combining table 13 and table 14, the synergist 1 significantly inhibits the germination of beauveria bassiana spores, the synergist 2 has little influence on the germination of beauveria bassiana spores, and has significant synergistic effect on pyrethrin, and the mass fraction of the synergist is preferably 5-10 times of the mass fraction of the pyrethrin (namely 2% -4%).
To sum up the test results of the embodiments 1 to 5, the dispersible oil suspension agent of pyrethrin and beauveria bassiana provided by the invention has 1 to 2 percent of effective components (including 0.3 to 0.8 percent of natural pyrethrin and 0.7 to 1.5 percent of beauveria bassiana conidium powder) by mass percentage; emulsifier 10-13% (including castor oil polyoxyethylene (40) ether 7-8%, polyoxyethylene 400 monooleate 2-3%, polyoxyethylene (40) perennial sorbate 1-2%); 7 to 8 percent of aqueous dispersant (polyoxyethylene sorbitol hexaoleate); 3 to 5 percent of non-aqueous dispersant (comprising 2 to 4 percent of star-shaped structure copolymer, 0 to 1 percent of alkoxy ethylene propylene sorbitol ester and 0 to 1 percent of polyalkylene block copolymer); 3-5% of a rheology modifier (2-3% of organobentonite, 1% of a polyester block copolymer, 1% of fumed silica); 2 to 4 percent of synergist (synergistic amine) and 0.005 to 0.01 percent of ultraviolet protective agent (TiO) 2 ) And the balance of oil base (vegetable blend oil, soybean oil, sunflower seed oil or peanut oil).
Example 6:1% of dispersible oil suspending agent of pyrethrin beauveria bassiana, the specific components are shown in table 15:
watch 15
The preparation method of the dispersible oil suspending agent containing 1% of pyrethrin and beauveria bassiana comprises the following steps:
(1) Weighing the components in proportion;
(2) Heating and dissolving the star-structure copolymer in a small amount of vegetable blend oil, adding about half of soybean oil for later use, and addingIn organic bentonite and TiO 2 (ii) a Adding emulsifier, non-aqueous dispersant and synergistic amine, mixing beauveria bassiana spore powder with a small amount of soybean oil, adding, stirring at 4000r/min with homogenizing emulsifier, adding pyrethrin and other rest components, adding plant blend oil to supplement the oil, and sanding to obtain 1% pyrethrin-beauveria bassiana dispersible oil suspension.
Example 7: the dispersible oil suspending agent of 2 percent pyrethrin beauveria bassiana comprises the following specific components:
TABLE 16
The preparation method of the dispersible oil suspension containing 2% pyrethrin and beauveria bassiana is the same as that of example 6.
Comparative example 1:1.5% pyrethrin beauveria bassiana dispersible oil suspending agent, the dosage of each component is shown in table 18:
watch 18
The preparation method of the 1.5% pyrethrin beauveria bassiana dispersible oil suspending agent is the same as that of example 6.
Comparative example 2:1% pyrethrin-beauveria bassiana dispersible oil suspending agent, and the dosage of each component is shown in table 19:
watch 19
The preparation method of the 1% pyrethrin beauveria bassiana dispersible oil suspending agent is the same as that of example 6.
Test example 1: quality detection of pyrethrin-beauveria bassiana dispersible oil suspending agent product
The dispersible oil suspension (muscardine silkworm OD) products of the pyrethrin beauveria bassiana prepared in the examples 6 and 7, the comparative examples 1 and the comparative examples 2 are stored in a standing way at room temperature (15-35 ℃) and low temperature (0-4 ℃), and indexes such as the fungus content, the viable bacteria rate, the pyrethrin content, the oil precipitation rate, the suspension rate, the pourability, the storage stability and the like of the beauveria bassiana are measured at regular intervals according to relevant standards.
The results show that the indexes of 1% and 2% of the chrysanthemin muscardine OD corresponding to the examples 6 and 7 are good (tables 20 and 21), and both meet the requirements of relevant standards; when stored under the conditions of heat (30 +/-2 ℃) for 18 weeks and cold (0 +/-2 ℃ and 1 week), the viable bacteria rate of beauveria bassiana of the comparative example 1 and the comparative example 2 is less than 90 percent and is obviously lower than that of the example 6 and the example 7.
TABLE 20 quality detection of pyrethrin-Beauveria bassiana dispersible oil suspending agent product
Product quality detection method for dispersible oil suspending agent containing pyrethrin and beauveria bassiana in TABLE 21
FIG. 2 is the appearance of four dispersible oil suspensions of examples 6 and 7 and comparative examples 1 and 2 after hot storage; wherein fig. 2A is the dispersible oil suspension prepared in example 6, fig. 2B is the dispersible oil suspension prepared in example 7, fig. 2C is the dispersible oil suspension prepared in comparative example 1, and fig. 2D is the dispersible oil suspension prepared in comparative example 2.
Test example 2: indoor biological activity assay
Test aphids: melon aphids (Aphis gossypii Glover) collected from the Experimental farm of southern China university of agriculture are bred on healthy cucumbers, stably bred for many generations for experiment, kept under proper illumination, humidity and temperature, and selected healthy adults for bioactivity determination.
Reagent to be tested: in the present invention, 1.0% of the chrysanthemin muscardine OD (containing 0.3% of natural pyrethrin and 0.7% of muscardine spore powder) of example 6, 2.0% of the chrysanthemin muscardine OD (containing 0.5% of natural pyrethrin and 1.5% of muscardine spore powder) of example 7, and 1.5% of the natural pyrethrin aqueous emulsion (yunnan nan bao biotechnology ltd.); 1.5% pyrethrin-beauveria bassiana dispersible oil suspending agent of comparative example 1 (containing 0.5% natural pyrethrin and 1% beauveria bassiana spore powder), and 1% pyrethrin-beauveria bassiana dispersible oil suspending agent of comparative example 2 (containing 0.3% natural pyrethrin and 0.7% beauveria bassiana spore powder).
The experimental method and the steps are as follows: the biological activity of the medicament on aphids is measured by using a spraying method, the aphids are inoculated into healthy cucumber pot plants 3 days in advance, and the aphids are reserved after being stably propagated. Diluting the test agent with water 1000 times, 2000 times, 3000 times, 5000 times, 7000 times and 10000 times respectively for use. The prepared liquid medicine is evenly sprayed on cucumber leaves with aphids, the cucumber leaves are moistened without dripping water, and the control group is sprayed with sterile water and repeated for 3 times. Culturing in a light incubator under the conditions of light 14 (L: D), temperature 25 +/-1 ℃ and relative humidity 70 +/-10%. Aphid death was observed every 24h and was determined as death with immobility of the extremities of aphids. And (5) counting the mortality rate and correcting the mortality rate. The obtained data are subjected to variance analysis and Probit toxicity regression analysis by SPSS software.
TABLE 22 Activity of Agents on aphids of melon
Note: the data in the table are mean ± sd, and different letters after the same column indicate significant differences at P < 0.05 level (new repolarization test, DMRT).
The results are shown in Table 22, LC of two Chrysin Beauveria bassiana OD formulations of examples 6 and 7 50 The values are obviously lower than those of the comparison example 1, the comparison example and the 1.5% pyrethrin aqueous emulsion, which shows that the formula of the pyrethrin beauveria bassiana dispersible oil suspension (the beauveria bassiana OD) provided by the invention has more excellent aphid killing activity.
In conclusion, compared with a contrast medicament, the pyrethrin-beauveria bassiana dispersible oil suspending agent provided by the invention has the advantages that the medicament effect on aphids is obviously improved, the consumption of natural pyrethrin is obviously reduced, and the production cost is reduced.
Claims (10)
1. The dispersible oil suspending agent containing pyrethrin and beauveria bassiana is characterized in that the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 10 to 13 percent of emulsifier, 7 to 8 percent of aqueous dispersant, 3 to 5 percent of non-aqueous dispersant, 3.5 to 5 percent of rheological modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
2. The dispersible oil suspending agent containing pyrethrin and beauveria bassiana of claim 1, wherein the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 to 0.8 percent of natural pyrethrin, 0.7 to 1.5 percent of conidia of beauveria bassiana, 10 to 12 percent of emulsifier, 7.5 percent of aqueous dispersant, 4 percent of non-aqueous dispersant, 4 to 5 percent of rheology modifier, 2 to 4 percent of synergist, 0.005 to 0.01 percent of ultraviolet protective agent and the balance of oil base.
3. The dispersible oil suspending agent containing pyrethrin and beauveria bassiana of claim 1, wherein the dispersible oil suspending agent is prepared from the following raw materials in percentage by mass: 0.3 percent of natural pyrethrin, 0.7 percent of beauveria bassiana conidia, 11 percent of emulsifier, 7.5 percent of aqueous dispersant, 4 percent of non-aqueous dispersant, 4 percent of rheology modifier, 3 percent of synergist, 0.007 percent of ultraviolet protective agent and 69.493 percent of oil base.
4. The dispersible oil suspension concentrate comprising pyrethrin and beauveria bassiana of any of claims 1-3, wherein said emulsifier is a non-ionic surfactant;
the aqueous dispersant comprises at least one of polyoxyethylene ether dispersant and polyoxyethylene (40) sorbitol hexaoleate;
the non-aqueous dispersant is at least one of an alkoxyethylene (5) propylene (15) based sorbitol ester, a polyalkylene block copolymer and a star structure copolymer;
the rheology modifier comprises at least one of diatomite, xanthan gum, sodium carboxymethylcellulose, organobentonite, a polyester block copolymer and fumed silica;
the synergist is synergist amine;
the ultraviolet protective agent comprises at least one of zinc oxide, 2- (3, 5-di-tert-amyl-2-hydroxyphenyl) benzotriazole and titanium dioxide.
The oil base comprises at least one of sunflower oil, soybean oil, peanut oil and vegetable blend oil.
5. The dispersible oil suspension concentrate of pyrethrin and beauveria bassiana of claim 4, wherein said non-ionic surfactant comprises at least one of polyoxyethylene (40) sorbitan hexaoleate, polyalkylene block copolymers, castor oil polyoxyethylene ether castor oil polyoxyethylene (10) ether, castor oil polyoxyethylene ether castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene (30) ether, castor oil polyoxyethylene (40) ether, castor oil polyoxyethylene ether, alkoxy ethylene (5) propylene (15) sorbitan ester, polyalkylene block copolymers, star copolymers, polyethylene glycol 400 monooleate, and polyoxyethylene (40) sorbitan monooleate.
6. The dispersible oil suspension concentrate of pyrethrin and beauveria bassiana of claim 5, wherein said non-ionic surfactant comprises castor oil polyoxyethylene (40) ether, polyethylene glycol 400 monooleate, and polyoxyethylene (40) sorbitan perester;
the mass ratio of the castor oil polyoxyethylene (40) ether to the polyethylene glycol 400 monooleate to the polyoxyethylene (40) perennial sorbitol ester is (7-8): (2-3): (1-2).
7. The dispersible oil suspension concentrate comprising pyrethrin and beauveria bassiana of claim 4, wherein said polyoxyethylene ether-based dispersant is a polyalkylene block copolymer;
the aqueous dispersant comprises polyoxyethylene (40) sorbitol hexaoleate;
the non-aqueous dispersant comprises a star-structure copolymer, an alkoxyethylene (5) propylene (15) sorbitol ester and the non-ionic block copolymer, wherein the mass ratio of the star-structure copolymer, the alkoxyethylene (5) propylene (15) sorbitol ester and the non-ionic block copolymer is (3-5): (0-1): (0-2).
8. The dispersible oil suspension concentrate of claim 4 comprising pyrethrin and beauveria bassiana,
the rheology modifier comprises organobentonite, a polyester block copolymer and fumed silica;
the mass ratio of the organic bentonite to the polyester block copolymer to the fumed silica is (2-3): 1;
the vegetable blend oil comprises soybean oil, peanut oil and sesame oil; the mass ratio of the soybean oil to the peanut oil to the sesame oil is 90.
9. The use of the dispersible oil suspension concentrate comprising pyrethrin and beauveria bassiana as claimed in any of claims 1 to 8 for controlling agricultural pests.
10. The use according to claim 9, wherein the agricultural pest is a homopteran pest.
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