Preparation method and application of chicken manure biogas slurry purification and refining component

Abstract

The invention provides a preparation method and application of a chicken manure biogas slurry purified and refined component, the purified and refined component prepared by the method is a phytotoxicity relieving product which is wide in crop phytotoxicity application range and friendly to the ecological environment, and an application method capable of achieving phytotoxicity relieving and pesticide effect increasing on crops in multiple ways and at different time can be achieved. The two-way functional effects of relieving phytotoxicity and enhancing the pesticide effect are realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of resource utilization of breeding waste and agricultural production, and particularly relates to a preparation method and application of a chicken manure biogas slurry purification and refining component. BACKGROUND

[0002] Pesticides are important inputs in agricultural production, and play an important role in the prevention and control of pests and weeds, and chemical weeding. In the actual agricultural production practice, excessive application of pesticides often occurs due to improper human operation, causing crop phytotoxicity stress and phytotoxicity, mainly including crop wilting, leaf abscission, leaf scorching and curling, growth stagnation and even death, and crop physiological and biochemical indexes mainly manifesting in a sharp increase in the content of antioxidant enzymes and malondialdehyde in plant cells. At present, there are various phytotoxicity relief products on the market, among which the main components of plant growth regulator products are brassinolide, indole acetic acid, sodium complex nitrophenol and other single or composite plant growth regulator phytotoxicity relief substances. However, plant hormone products have strong specificity for phytotoxicity, and the application range of a single product is narrow. In the process of application, the dosage and concentration need to be highly concerned. The conventional phytotoxicity relief agents are applied after pesticide application, and the time node is difficult to grasp. Once the growth point of the crop is damaged by the pesticide, the phytotoxicity relief agent cannot play an effective role. If the concentration and timing are not appropriate, not only the efficacy of the phytotoxicity relief agent is affected, but also further hormone excess hazards are caused. The phytotoxicity relief products of chloroacetamides, chloroacetamides and chloroacetamides mainly reduce the influence of phytotoxicity by specifically inducing crop detoxification metabolism mechanism, and are mainly used to solve the phytotoxicity problem of herbicides. However, they have no obvious effect on the phytotoxicity of other pesticides. At the same time, the chemical safeners inevitably bring new potential environmental pollution risks or crop safety hazards in the process of synthesis and use.

[0003] In addition, in the prior art, there are few phytotoxicity relief products that can simultaneously increase pesticide efficacy and reduce pesticide use. The method for enhancing the herbicidal effect of flupicolide and relieving phytotoxicity function proposes to use a plant hormone combination to increase the herbicidal effect of flupicolide and reduce phytotoxicity. The selection of pesticides has specificity, and is limited to a single form of stem and leaf spraying. Therefore, there is an important need to develop a phytotoxicity relief product with a wide application range for crops, which is friendly to the ecological environment, reduces the influence of crop phytotoxicity, and ensures the normal growth of phytotoxicity crops. SUMMARY

[0004] In view of the above problems, the application provides a preparation method of a purified and refined component of chicken manure biogas slurry, based on the specificity of component material composition, and through a large number of tests and tests, the technical personnel find that different application methods can play a dual role of drug damage and drug effect, the application further innovates the application method of drug damage and drug effect in multiple ways and at different times on crops on the basis of innovating the purification and refining method of chicken manure biogas slurry, realizes the dual functional role of drug damage and drug effect, and has outstanding application value and obvious creativity.

[0005] To achieve the above object, the specific technical scheme provided by the application is as follows:

[0006] The first aspect of the application provides a preparation method of a purified and refined component of chicken manure biogas slurry, the preparation method comprising: fully anaerobic fermentation of chicken manure to obtain original biogas slurry material; the original biogas slurry material is transported into an acidification adjustment tank, air is introduced for acidification hydrolysis, an ultrasonic wave generating device is arranged at the output pipeline of the acidification adjustment tank, and ultrasonic wave treatment is continuously performed; the biogas slurry after acidification hydrolysis and ultrasonic wave treatment is transported to an ultrafiltration raw material tank for ultrafiltration treatment to obtain an ultrafiltration clear liquid; the ultrafiltration clear liquid is subjected to nanofiltration treatment to obtain a nanofiltration concentrated liquid; the nanofiltration concentrated liquid is subjected to dewatering treatment to obtain a primary biogas slurry concentrated component; anhydrous methanol is added to the primary biogas slurry concentrated component, and ultrasonic wave is combined for leaching separation, anhydrous methanol is leached for 3-5 times, and methanol extraction liquid is separated and collected; the methanol extraction liquid is combined and subjected to reduced pressure distillation to obtain a methanol extract, water is added for redissolution, ethyl acetate is extracted to obtain ethyl acetate extraction liquid, and the extraction is performed for 3-5 times; after the second ethyl acetate extraction is completed, hydrochloric acid is added to the water phase solution separated and collected for pH adjustment, and ethyl acetate solvent extraction is further performed; the ethyl acetate extraction liquid extracted for multiple times is combined and subjected to reduced pressure distillation to obtain an ethyl acetate extract, a 50% methanol solution is added for redissolution to obtain the purified and refined component.

[0007] In some embodiments, the acidification hydrolysis is performed at 25-30 DEG C for 24 h.

[0008] Further, the acidification hydrolysis is performed at 25 DEG C for 24 h.

[0009] In some embodiments, the ultrasonic wave treatment is 20-25 kHz ultrasonic wave treatment.

[0010] Further, the ultrasonic wave treatment is 25 kHz ultrasonic wave treatment.

[0011] In some embodiments, the ultrafiltration pore size of the ultrafiltration is 100-200 nm.

[0012] Further, the ultrafiltration pore size of the ultrafiltration is 100 nm.

[0013] Further, the nanofiltration pore size is 1 nm.

[0014] In some embodiments, the dehydration process is as follows: the nanofiltration concentrated solution is pumped into a reaction kettle for primary dehydration treatment, low-pressure distillation, vacuum degree is lower than -0.09 Mpa, 30-35℃, to obtain concentrated solution; the concentration multiple is 5-10 times; the concentrated solution obtained by low-pressure distillation is subjected to secondary freeze-drying dehydration treatment by a freeze dryer, freeze-drying treatment parameters: pre-freezing temperature -25--20℃; drying temperature 30-35℃, pressure <100 pa.

[0015] Further, the dehydration process is as follows: the nanofiltration concentrated solution is pumped into a reaction kettle for primary dehydration treatment, low-pressure distillation, vacuum degree is lower than -0.09 Mpa, 35℃, to obtain concentrated solution; the concentration multiple is 10 times; the concentrated solution obtained by low-pressure distillation is subjected to secondary freeze-drying dehydration treatment by a freeze dryer, freeze-drying treatment parameters: pre-freezing temperature -25℃; drying temperature 30℃, pressure <100 pa.

[0016] In some embodiments, the leaching separation process is as follows: the primary biogas liquid concentrated component is placed in a reaction kettle with an ultrasonic device, anhydrous methanol is added according to a mass ratio of 1:4-1:8, the ultrasonic wave parameters are 25-30 kHz, and each leaching time is 1-2 h; the centrifugal parameters of the leaching liquid are 1000 g, and the centrifugation time is 20-30 min.

[0017] Further, the leaching separation process is as follows: the primary biogas liquid concentrated component is placed in a reaction kettle with an ultrasonic device, anhydrous methanol is added according to a mass ratio of 1:8, the ultrasonic wave parameters are 25-30 kHz, and each leaching time is 1-2 h; the centrifugal parameters of the leaching liquid are 1000 g, and the centrifugation time is 20-30 min.

[0018] In some embodiments, the vacuum distillation conditions of the methanol extract liquid are as follows: vacuum degree is lower than -0.09 Mpa, 30-35℃.

[0019] Further, the vacuum distillation conditions of the methanol extract liquid are as follows: vacuum degree is lower than -0.09 Mpa, 35℃.

[0020] In some embodiments, the water addition and resolubilization ratio is water to methanol extract mass ratio 1:1-1:1.5.

[0021] Further, the water addition and resolubilization ratio is water to methanol extract mass ratio 1:1.5.

[0022] In some embodiments, the ethyl acetate extraction ratio is ethyl acetate to methanol extract resolubilization solution weight ratio 1:1-1:4.

[0023] Further, the ratio of the ethyl acetate extraction is ethyl acetate to methanol extract re-dissolution solution weight ratio 1:4.

[0024] Further, the process of the ethyl acetate extraction is as follows: oscillation 0.5-1 h, standing 1 h, separating the ethyl acetate phase by a liquid separator.

[0025] In some embodiments, after the end of the second ethyl acetate extraction, 1 mol / L hydrochloric acid is added to the water phase solution separated by extraction to adjust the pH to 2.5-3.0, and then ethyl acetate solvent extraction is continued, ethyl acetate is added at a water phase solution mass ratio of 1:1-1:4, oscillation is performed for 0.5-1 h, standing is performed for 1 h, the ethyl acetate phase is separated by a liquid separator, and extraction is continued for 1-3 times.

[0026] Further, after the end of the second ethyl acetate extraction, 1 mol / L hydrochloric acid is added to the water phase solution separated by extraction to adjust the pH to 3.0, and then ethyl acetate solvent extraction is continued, ethyl acetate is added at a water phase solution mass ratio of 1:4, oscillation is performed for 0.5 h, standing is performed for 1 h, the ethyl acetate phase is separated by a liquid separator, and extraction is continued for 2 times.

[0027] In some embodiments, the reduced pressure distillation condition of the ethyl acetate extraction solution is that the vacuum degree is lower than -0.09 Mpa, and the temperature is 30-40℃.

[0028] Further, the reduced pressure distillation condition of the ethyl acetate extraction solution is that the vacuum degree is lower than -0.09 Mpa, and the temperature is 35℃.

[0029] In some embodiments, the re-dissolution ratio of the 50% methanol solution is 50% methanol solution to ethyl acetate extract mass ratio 1:1-1:5.

[0030] Further, the re-dissolution ratio of the 50% methanol solution is 50% methanol solution to ethyl acetate extract mass ratio 1:2.5.

[0031] The second aspect of the present application provides a purified and refined component prepared by the preparation method according to the first aspect of the present application.

[0032] The third aspect of the present application provides a method for relieving pesticide phytotoxicity, which comprises applying the purified and refined component according to the second aspect of the present application.

[0033] Further, the pesticide comprises herbicides, insecticides, and fungicides.

[0034] Further, the pesticide is nicosulfuron.

[0035] In a specific embodiment of the present invention, nicosulfuron, also known as Yunolu or nicosulfuron, is a selective herbicide of the sulfonylurea class, with the molecular formula C. 15 H 18 N6O6S.

[0036] Furthermore, the pesticide is 2,4-D isooctyl ester.

[0037] In a specific embodiment of the present invention, 2,4-D isooctyl ester refers to ester with the molecular formula C 16 H 22 Cl2O3 is a selective post-emergence foliar contact herbicide suitable for soybeans, corn, and wheat.

[0038] In some embodiments, the method includes applying the purified and refined components described in the second aspect of the invention prior to sowing.

[0039] Furthermore, the method of applying the purified and refined components is to soak the seeds after dilution.

[0040] Furthermore, the dilution factor is 10,000 to 50,000 times.

[0041] Furthermore, the dilution factor is 10,000 times.

[0042] Furthermore, the soaking temperature is 25~30℃.

[0043] Furthermore, the soaking time is 8-12 hours.

[0044] In some embodiments, the method includes applying the purified and refined component described in the second aspect of the invention prior to the application of pesticides.

[0045] Furthermore, the purified and refined components are applied by foliar spraying after dilution.

[0046] Furthermore, the dilution factor is 20,000 to 100,000 times.

[0047] Furthermore, the dilution factor is 20,000 times.

[0048] Furthermore, the purified and refined components are applied 2 to 3 times, with an interval of 3 to 5 days.

[0049] Furthermore, the method for applying the purified and refined components is as follows: before applying the pesticide, dilute the purified and refined components and spray them on the leaves, with an application interval of 3 to 5 days, and apply them more than twice in a row, and then apply the pesticide.

[0050] Furthermore, the interval between the last application of the purified and refined component and the application of the pesticide should be within 48 hours.

[0051] In some embodiments, the method includes applying the purified and refined component described in the second aspect of the invention after crop phytotoxicity has occurred.

[0052] Furthermore, the purified and refined components are applied by dilution and then sprayed onto the stems and leaves.

[0053] Furthermore, the dilution factor is 20,000 to 100,000 times.

[0054] Furthermore, the dilution factor is 50,000 times.

[0055] Furthermore, the purified and refined components are applied 2 to 3 times, with an interval of 1 to 3 days.

[0056] Furthermore, the method for applying the purified and refined components is as follows: after the crop suffers phytotoxicity, the purified and refined components are diluted and sprayed on the stems and leaves, with an application interval of 1 to 3 days, and applied more than twice consecutively.

[0057] Furthermore, the purified and refined components are applied at the initial stage of drug-induced damage symptoms.

[0058] Furthermore, the purified and refined components are applied within 24 hours of the onset of phytotoxicity symptoms.

[0059] Furthermore, the purified and refined components are used to remove severely phytotoxic tissues from crops before application.

[0060] The fourth aspect of the present invention provides a method for enhancing the efficacy of pesticides, the method comprising mixing and applying the purified and refined components described in the second aspect of the present invention with a pesticide.

[0061] Furthermore, the pesticides include herbicides, insecticides, and fungicides.

[0062] Furthermore, the pesticide in question is tebuconazole.

[0063] In a specific embodiment of the present invention, tebuconazole is an organic compound with the molecular formula C. 16 H 22 ClN3O is a highly effective, broad-spectrum, systemic triazole fungicide, mainly used to control various fungal diseases on crops such as wheat, rice, peanuts, vegetables, bananas, apples, pears, corn, and sorghum.

[0064] Furthermore, the method of mixed application is as follows: the purified and refined components are diluted, mixed with pesticides, and applied to the soil surface and / or foliar spray; the amount of pesticide used is reduced by 10% to 20% compared to the method without purified and refined components.

[0065] Furthermore, the dilution factor is 20,000 to 100,000 times.

[0066] Furthermore, the dilution factor is 20,000 times, and the amount of pesticide used is reduced by 20% compared to the unrefined component.

[0067] The fifth aspect of the present invention provides the application of the purified and refined components described in the second aspect of the present invention, the application including any one of the following:

[0068] 1) Application in eliminating pesticide residue damage in soil before sowing; 2) Application in eliminating pesticide damage in soil after sowing; 3) Application in preventing pesticide damage before applying pesticides to crops; 4) Application in promoting recovery of crops after pesticide damage symptoms appear; 5) Application in enhancing pesticide efficacy and reducing pesticide usage.

[0069] Furthermore, the pesticides include herbicides, insecticides, and fungicides.

[0070] Furthermore, the pesticide in question is nicosulfuron.

[0071] Furthermore, the pesticide is 2,4-D isooctyl ester.

[0072] Furthermore, the pesticide in question is tebuconazole.

[0073] Furthermore, the crops are selected from field crops and cash crops.

[0074] Furthermore, the crop in question is corn.

[0075] Furthermore, the crop in question is an apple.

[0076] Advantages and beneficial effects of the present invention:

[0077] 1) The purified and refined components provided by this invention are derived from anaerobic fermentation biogas slurry and are prepared through multi-stage purification and refining, exhibiting biosafety. The extraction conditions in the purification and refining process were optimized through long-term comparative testing, and all selected extractants are recyclable and reusable, making the preparation process environmentally friendly.

[0078] 2) The purified and refined components provided by this invention integrate the effects of relieving drug toxicity and enhancing drug efficacy through innovation and optimization of the application method.

[0079] 3) The purified and refined components provided by this invention can be used to relieve pesticide damage in multiple scenarios and through multiple pathways. The application methods for relieving pesticide damage before seed soaking, before application, and after pesticide damage occurs are more practical and can ensure the needs of crops for relieving pesticide damage in multiple scenarios. Detailed Implementation

[0080] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0081] Example 1: Preparation of specific chicken manure biogas slurry concentration, purification and refining components

[0082] a. Chicken manure undergoes thorough mesophilic anaerobic fermentation to obtain raw biogas slurry.

[0083] b. The raw biogas slurry is transported to an acidification and equalization tank, where air is continuously introduced for 24 hours of acidification and hydrolysis at 25°C, achieving hydrolysis and sedimentation of the raw biogas slurry. An ultrasonic generator is installed at the output pipe of the acidification and equalization tank to continuously perform 25kHz ultrasonic treatment, promoting the degradation of insoluble or sparingly soluble substances in the biogas slurry and further converting large molecules into smaller molecules, facilitating the subsequent separation and collection of smaller molecules by the membrane.

[0084] c. The biogas slurry, after acidification, hydrolysis, and ultrasonic treatment, is transported to an ultrafiltration feed tank, where it undergoes ultrafiltration through a membrane, and the ultrafiltrate is collected. The ultrafiltrate is further concentrated and separated using a nanofiltration membrane, and the nanofiltration concentrate is collected. Specifically, the ultrafiltration pore size is 100 nm; the nanofiltration pore size is 1 nm.

[0085] d. The nanofiltration concentrate is dehydrated to obtain the primary biogas slurry concentrate. Specifically, the nanofiltration concentrate is pumped into a reactor for primary dehydration, followed by low-pressure distillation at a vacuum below -0.09 MPa and 35°C to obtain a concentrated solution. The concentration factor is 10 times. The concentrated solution obtained from low-pressure distillation is then subjected to secondary freeze-drying dehydration using the following parameters: pre-freezing temperature -25°C; drying temperature 30°C; pressure <100 Pa, to obtain the primary biogas slurry concentrate.

[0086] e. Add anhydrous methanol to the primary biogas slurry concentrate and perform extraction and separation using ultrasound. Collect the filtrate after methanol extraction. Specifically, place the primary biogas slurry concentrate in a reaction vessel (equipped with an ultrasonic device), add anhydrous methanol at a mass ratio of 1:8, use ultrasonic parameters of 25-30 kHz, and extract for 1-2 hours each time. Centrifuge the extract at 1000g for 20-30 minutes. Repeat the anhydrous methanol extraction three times.

[0087] f. Combine the methanol extracts and subject them to vacuum distillation to remove methanol, obtaining a methanol extract. Vacuum distillation parameters: vacuum level below -0.09 MPa, 35°C. The recovered methanol can be reused for extraction.

[0088] g. Redissolve the methanol extract in water. Specifically, redissolve in water at a weight ratio of 1:1.5 with the methanol extract.

[0089] h. Add pure ethyl acetate to the reconstituted methanol extract and shake to promote extraction. Allow to stand and separate the ethyl acetate phase. Specifically, take the reconstituted methanol extract and add ethyl acetate at a weight ratio of 1:4. Shake for 0.5 hours each time, let stand for 1 hour, and separate the ethyl acetate phase using a separatory device. Repeat the extraction 3-5 times.

[0090] i. After the second ethyl acetate extraction, adjust the pH to 3.0 by adding 1 mol / L hydrochloric acid to the aqueous phase solution from the extraction separation, and then continue the ethyl acetate solvent extraction. Add ethyl acetate at a weight ratio of 1:4 with the aqueous phase solution, shaking for 0.5 h each time, letting stand for 1 h, and separating the ethyl acetate phase using a separatory device. Repeat the extraction twice. Combine the obtained ethyl acetate extract with the ethyl acetate extracts obtained from the first two extractions.

[0091] According to the experimental results, after two extractions of ethyl acetate, the pH was adjusted to 3.0 with hydrochloric acid, and then the ethyl acetate was extracted twice more. The final combined ethyl acetate extract weight was 18% higher than that of four extractions without hydrochloric acid adjustment (p < 0.05).

[0092] j. Combine the multiple ethyl acetate extracts and perform vacuum distillation to remove ethyl acetate. The vacuum distillation parameters are below -0.09 MPa and 35°C. The recovered ethyl acetate can be reused for extraction.

[0093] k. Collect the ethyl acetate extract, and redissolve the extract in 50% methanol solution at a weight ratio of 1:2.5 with the ethyl acetate extract. Collect the solution, which is the concentrated, purified and refined component of chicken manure biogas slurry.

[0094] Example 2: Application method and efficacy test of purified and refined components in crop pesticide damage relief.

[0095] Application Method 1: Apply by soaking seeds. This avoids situations where excessive pesticide residues in the previous crop soil negatively impact seed germination or seedling growth, ensuring normal crop growth. The specific application method is as follows: Before sowing in soil with excessive pesticide residues from the previous crop, dilute the purified and refined components 10,000-50,000 times, soak the seeds at 25℃ for 12 hours, and then sow. This method is also suitable for situations where pesticides need to be sprayed on the soil surface after sowing, preventing excessive pesticides from affecting seed germination or seedling growth and ensuring normal crop growth. Pesticides include herbicides, insecticides, and fungicides.

[0096] Efficacy verification: Indoor test on the effect of purified and refined components in seed soaking to reduce nicosulfuron-induced herbicide damage to maize.

[0097] 1. Experimental materials: maize variety “Denghai 605”; 4% nicosulfuron; purified and refined components; raw biogas slurry.

[0098] 2. Test methods

[0099] 1) Seed soaking: The purified and refined components were prepared into solutions diluted 100,000 times, 50,000 times, 20,000 times, 10,000 times, and 5,000 times with deionized water, respectively; the original biogas slurry was prepared into a solution diluted 100 times with deionized water (based on the characteristics of high-concentration chicken manure biogas slurry, a 100-fold dilution is suitable for seed soaking treatment); deionized water was used as a control. Seeds were soaked at a dosage of 2 L of solution per kilogram of corn seeds for 12 hours at 25℃.

[0100] 2) Preparation of medicated soil: Use soil from vegetable greenhouses and set the concentration of nicosulfuron in the soil to 0.25 mg / kg. First, dilute 4% nicosulfuron 16,000 times to prepare a 2.5 mg / L solution. Then, add 100 mL of the solution to each kilogram of soil, mix well, and let it stand in a cool place for 24 hours.

[0101] 3) Sowing: Each experimental group was set up with 3 trays. The medicated soil was used as the seedling substrate. The soaked seeds were sown into the 32-cell seedling trays, one seed per cell, and the seeds were gently pressed into the substrate to a depth of 1 cm with tweezers.

[0102] 4) Cultivation: Place the seedling trays in a light incubator and cultivate them at 30℃ / 25℃ under 14h light / 10h darkness conditions.

[0103] 5) Measurement indicators: The germination rate was recorded 6 days after sowing, and the plant height and fresh weight of the above-ground parts were measured 15 days later.

[0104] 3. Test treatment

[0105] Table 1. Indoor test groupings for reducing nicosulfuron-induced herbicide damage to maize after seed soaking

[0106]

[0107] 4. Test Results

[0108] Table 2. Results of laboratory tests on the reduction of nicosulfuron-induced herbicide damage to maize by seed soaking.

[0109]

[0110] The letters in the table represent the results of the difference analysis between different experimental groups. The same letter indicates no significant difference, while different letters indicate a significant difference.

[0111] The results are shown in Table 2. Regarding germination rate and plant height, the T3-T5 groups (corn seeds treated with a 50,000-10,000 times dilution of the purified component) showed significant improvements compared to the water control CK1 (soaked in water and then sprayed with nicosulfuron), but no significant difference compared to the blank control CK0. Regarding plant fresh weight, the T3-T5 groups (corn seeds treated with a 50,000-10,000 times dilution of the purified component) also showed significant improvements compared to the water control CK1 (soaked in water and then sprayed with nicosulfuron), and no significant difference compared to the blank control CK0, indicating an antidote effect. The T1 group (raw biogas slurry treatment) showed no significant differences in germination rate, plant fresh weight, and plant height compared to CK1, and did not show an antidote effect. In summary, the purified and refined components prepared according to the method of this patent, diluted 50,000 to 10,000 times, can be used to treat corn seeds by soaking them in water. This can effectively alleviate the phytotoxicity of corn caused by the herbicide nicosulfuron sprayed on the soil surface before emergence.

[0112] Application Method Two: Apply the purified and refined component by spraying before pesticide application. This pretreatment helps reduce the phytotoxic effects of subsequent pesticide application. The specific application method is as follows: Dilute the purified and refined component to 20,000 to 100,000 times its original concentration, and then spray it onto the leaves. Apply each time at least twice, with an interval of 3-5 days between applications, before applying the pesticide. The optimal time interval between the last application of the purified and refined component and the pesticide application should be within 48 hours. This can reduce the phytotoxic effects of subsequent pesticide applications on crops. Pesticides include herbicides, insecticides, and fungicides.

[0113] Efficacy verification: Application test of purified and refined components in preventing phytotoxicity of corn by 2,4-D isooctyl ester

[0114] 1. Experimental conditions: Maize (Denghai 605) was used as the test crop. All experimental groups had the same cultivation conditions and water, fertilizer and pesticide management.

[0115] 2. Experimental Treatments: After the third true leaf unfolded, spraying began with diluted raw biogas slurry and purified herbicides at different dilution ratios for each experimental group. Spraying was done twice, with a 3-day interval between applications. One day after the second spraying, a 500-fold dilution of 77% 2,4-D isooctyl ester herbicide was applied. Each experimental group had three replicates, with 60 plants per replicate. Phytotoxicity was assessed 5 days after 2,4-D isooctyl ester herbicide application, and the detoxification effect was calculated.

[0116] Table 3. Application test groups for preventing phytotoxicity of maize by 2,4-D isooctyl ester

[0117]

[0118] 3. Measurement Method

[0119] Phytotoxicity grading: Grade 0 - Normal growth, consistent with the control group in clean water; Grade 1 - Slightly deformed central leaves, no wilting of the plant; Grade 2 - Obviously deformed central leaves, slightly wilting of the plant; Grade 3 - Obviously deformed central leaves, mild wilting of the plant; Grade 4 - Severely deformed central leaves, obvious wilting of the plant; Grade 5 - Severe wilting of the plant, with the leaves turning severely yellow.

[0120]

[0121]

[0122] 4. Test Results

[0123] Table 4. Results of application trials for preventing herbicide damage to maize caused by 2,4-D isooctyl ester.

[0124]

[0125] The letters in the table represent the results of the difference analysis between different experimental groups. The same letter indicates no significant difference, while different letters indicate a significant difference.

[0126] The results are shown in Table 4. Spraying the purified and refined component twice before applying the herbicide during the corn seedling stage effectively prevented phytotoxicity caused by the herbicide 2,4-D isooctyl ester. The detoxification effect was highly influenced by the concentration of the purified and refined component, showing significant changes with different dilution ratios. Groups T3-T5 showed better detoxification effects, with virtually no obvious phytotoxicity, and their detoxification performance was significantly different from the control group (CK). Furthermore, their detoxification effects were significantly higher than those of groups T1, T2, and T6. Groups T1, T2, and T6, treated with biogas slurry, still exhibited significant phytotoxicity, including twisted and deformed leaf margins and wrinkled new leaves. In conclusion, spraying the purified and refined component prepared according to the method of this patent at a dilution of 100,000 to 20,000 times before applying 2,4-D isooctyl ester can effectively reduce the phytotoxicity caused by excessive 2,4-D isooctyl ester.

[0127] Application Method 3: Apply after pesticide damage occurs to reduce crop damage and promote rapid recovery of plant growth. The specific application method is as follows: Dilute the purified component to 20,000-100,000 times its original concentration and apply via foliar spray. Apply at 1-3 day intervals, repeating at least twice. This can reduce the impact of pesticide damage on crops and promote rapid plant recovery. Note: Application should be made at the initial stage of pesticide damage symptoms, preferably within 24 hours of the onset of symptoms. Before application, prune and remove severely damaged tissues of the plant.

[0128] Efficacy Verification: Application Trial of Purified Components to Alleviate Herbicide Damage to Corn by 2,4-D Isooctyl Ester

[0129] 1. Experimental conditions: Maize (Denghai 605) was used as the test crop. All experimental groups had the same cultivation conditions and water, fertilizer and pesticide management.

[0130] 2. Experimental Treatment: After the fourth true leaf unfolded, a 500-fold dilution of 77% 2,4-D isooctyl ester herbicide was sprayed on the leaves. On the first day after the appearance of phytotoxicity symptoms, spraying began with the original biogas slurry and purified components of the herbicide at different dilution ratios for each experimental group, with a two-day interval between sprays, for a total of two sprays. Each experimental group had three replicates, with 60 plants per replicate. Phytotoxicity was assessed five days after the second spraying of each experimental group, and the detoxification effect was calculated.

[0131] Table 5. Grouping of application trials to alleviate phytotoxicity of maize by 2,4-D isooctyl ester

[0132]

[0133] 3. Measurement Method

[0134] Phytotoxicity grading: Grade 0 - Normal growth, consistent with the control group in clean water; Grade 1 - Slightly deformed central leaves, no wilting; Grade 2 - Obviously deformed central leaves, slightly wilting; Grade 3 - Obviously deformed central leaves, mild wilting; Grade 4 - Severely deformed central leaves, obvious wilting; Grade 5 - Severe wilting, yellowing leaves.

[0135]

[0136]

[0137] 4. Test Results

[0138] Table 6. Results of application trials to alleviate herbicide damage to maize caused by 2,4-D isooctyl ester

[0139]

[0140] The results are shown in Table 6. After phytotoxicity occurred in corn, spraying the purified and refined component diluted twice could alleviate the phytotoxicity of the herbicide 2,4-D isooctyl ester on corn plants. The detoxification effect was highly affected by the concentration of the purified and refined component, showing significant changes in effect with different dilution ratios. Groups T3-T5 showed low phytotoxicity indices and good detoxification effects, with significant differences from the phytotoxicity index of the CK group. Group T1, treated with raw biogas slurry, showed no significant difference from the CK group and still exhibited obvious phytotoxicity symptoms. The detoxification effects of groups T3-T5 were significantly higher than those of groups T2, T6, and group T1 treated with raw biogas slurry. In summary, the purified and refined component prepared according to the method of this patent can effectively reduce the phytotoxicity of excessive 2,4-D isooctyl ester on crops and promote crop recovery after phytotoxicity caused by spraying the purified and refined component at a dilution of 100,000 to 20,000 times after spraying 2,4-D isooctyl ester.

[0141] Example 3: Application method and effect testing of purified and refined components for enhancing crop efficacy.

[0142] Application Method: When the purified and refined components are mixed with pesticides, the efficacy of the pesticides can be enhanced. These pesticides include those applied to the soil surface or as foliar sprays. The specific application method is as follows: Before mixing with the pesticide, dilute the purified and refined components 20,000 to 100,000 times, then mix with the diluted pesticide. It is important to reduce the pesticide dosage by 10% to 20% to achieve increased efficacy and reduced pesticide usage.

[0143] Efficacy verification: Field trial of synergistic effect of purified and refined components combined with tebuconazole

[0144] 1. Experimental conditions: The experimental site was located in the Fuji apple orchard in Nanwang Street, Penglai District, Yantai City, Shandong Province, with 10-year-old trees. All experimental plots had the same cultivation conditions, except for differences in the application of tebuconazole; other water, fertilizer, and pesticide management were consistent.

[0145] 2. Experimental Treatments: Following the orchard's pesticide application plan, spraying began before the onset of apple ring rot and was conducted monthly, specifically in June, July, August, and September (4 times in total). Each experimental group had 3 replicates, with 3 trees per replicate. The pesticide used was 43% tebuconazole suspension, with treatments at the standard dose and a 20% reduction (diluted 4000 and 5000 times, respectively) serving as control groups. The 43% tebuconazole suspension was mixed with either raw biogas slurry or a diluted purified component, forming the experimental groups. Additionally, groups using only diluted purified components (100,000, 50,000, and 20,000 times) were included to assess the effectiveness of the purified component.

[0146] Table 7. Field trial groups showing synergistic effects of purified components combined with tebuconazole

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[0148] 3. Measurement method: During the harvest period (October), select 2 trees in each plot, investigate all fruits, record the total number of fruits and the number of diseased fruits, and calculate the control effect based on the disease rate.

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[0151] 4. Test Results

[0152] Table 8. Field trial results of synergistic effect of purified components combined with tebuconazole

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[0154] The letters in the table represent the results of the difference analysis between different experimental groups. The same letter indicates no significant difference, while different letters indicate a significant difference.

[0155] The results are shown in Table 8. When the purified and refined components were mixed with the fungicide tebuconazole, the control effect of group T6 was not significantly different from that of group CK1 (concentrated with tebuconazole at the conventional dosage), and was significantly better than that of group CK2 (concentrated with tebuconazole at a 20% reduction). The control effects of groups T7 and T8 were significantly better than those of group CK1 (concentrated with tebuconazole at the conventional dosage) and group CK2 (concentrated with tebuconazole at a 20% reduction). When tebuconazole was reduced by 20%, and the mixture was applied with a 100,000-20,000 times dilution of the purified and refined components, the control effect could be achieved at the conventional dosage, or even exceed the effect of the conventional dosage.

[0156] The control effect of group T5 was not significantly different from that of group T9. Although it showed a certain synergistic effect with tebuconazole, it was significantly different from that of group CK1, which used tebuconazole at the conventional dosage, and did not effectively exert the synergistic effect of tebuconazole in controlling apple ring rot. The control effect of group T4 (raw biogas slurry) was not significantly different from that of group CK2 (tebuconazole dosage reduced by 20%), and did not show a synergistic effect. To further verify whether the purified and refined component itself has the effect of controlling apple ring rot, this experiment set up test groups for applying the purified and refined component at dilutions of 100,000 times, 50,000 times, and 20,000 times. The test results showed that applying the purified and refined component alone could not control apple ring rot. In summary, the purified and refined component prepared according to the method of this patent, diluted 100,000 to 20,000 times, and then mixed with tebuconazole can enhance the effect of tebuconazole in controlling apple ring rot, achieving the effect of synergistic effect and reduced dosage of pesticide.

[0157] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing purified and refined components of chicken manure biogas slurry, characterized in that, The preparation method includes: Chicken manure undergoes thorough mesophilic anaerobic fermentation to obtain raw biogas slurry. The raw biogas slurry material is transported to an acidification and conditioning tank, where air is introduced for acidification and hydrolysis. An ultrasonic generator is installed at the output pipe of the acidification and conditioning tank to continuously perform ultrasonic treatment. The biogas slurry after acidification, hydrolysis, and ultrasonic treatment is transported to the ultrafiltration feed tank for ultrafiltration treatment to obtain ultrafiltration clear liquid. The ultrafiltrate is subjected to nanofiltration to obtain nanofiltration concentrate; The nanofiltration concentrate is dehydrated to obtain a primary biogas slurry concentrate. Anhydrous methanol was added to the primary biogas slurry concentrate, and the extraction and separation were carried out by ultrasonic extraction. The anhydrous methanol was extracted 3 to 5 times, and the methanol extract was collected. The methanol extracts were combined and subjected to vacuum distillation to obtain a methanol extract. After redissolving in water, the extracts were extracted with ethyl acetate to obtain an ethyl acetate extract. The extraction was repeated 3 to 5 times. After the second ethyl acetate extraction, hydrochloric acid was added to the aqueous phase solution to adjust the pH, and ethyl acetate solvent extraction was continued. The ethyl acetate extracts obtained from multiple extractions were combined and subjected to vacuum distillation to obtain an ethyl acetate extract. The extract was then redissolved in 50% methanol solution to obtain the purified component.

2. The preparation method according to claim 1, characterized in that, The acidification and hydrolysis conditions are 25~30℃ for 24 hours; Preferably, the acidification and hydrolysis conditions are 25°C for 24 hours; Preferably, the ultrasonic treatment is a 20-25 kHz ultrasonic treatment; Preferably, the ultrasonic treatment is a 25 kHz ultrasonic treatment; Preferably, the ultrafiltration pore size is 100~200 nm; Preferably, the ultrafiltration pore size is 100 nm; Preferably, the nanofiltration pore size is 1 nm; Preferably, the dehydration process is as follows: the nanofiltration concentrate is pumped into a reaction vessel for primary dehydration, followed by low-pressure distillation at a vacuum level below -0.09 MPa and a temperature of 30~35°C to obtain a concentrated solution with a concentration ratio of 5~10 times; the concentrated solution obtained from low-pressure distillation is then subjected to secondary freeze-drying dehydration using a freeze dryer with the following freeze-drying parameters: pre-freezing temperature -25~-20°C; drying temperature 30~35°C; pressure <100 Pa. Preferably, the dehydration process is as follows: the nanofiltration concentrate is pumped into a reaction vessel for primary dehydration, followed by low-pressure distillation at a vacuum level below -0.09 MPa and a temperature of 35°C to obtain a concentrated solution with a concentration factor of 10 times; the concentrated solution obtained from low-pressure distillation is then subjected to secondary freeze-drying dehydration using a freeze dryer with the following freeze-drying parameters: pre-freezing temperature -25°C; drying temperature 30°C; pressure <100 Pa. Preferably, the extraction and separation process is as follows: the primary biogas slurry concentrate is placed in a reaction vessel equipped with an ultrasonic device, and anhydrous methanol is added at a mass ratio of 1:4 to 1:

8. The ultrasonic parameters are 25 to 30 kHz, and the extraction is carried out for 1 to 2 hours each time. The extract is centrifuged at 1000g for 20 to 30 minutes. Preferably, the extraction and separation process is as follows: the primary biogas slurry concentrate is placed in a reactor equipped with an ultrasonic device, anhydrous methanol is added at a mass ratio of 1:8, the ultrasonic parameters are 25~30 kHz, and the extraction is carried out for 1~2 hours each time; the extract is separated by centrifugation at 1000g for 20~30 minutes. Preferably, the reduced pressure distillation conditions for the methanol extract are: vacuum degree below -0.09 MPa, 30~35℃; Preferably, the reduced pressure distillation conditions for the methanol extract are: vacuum degree below -0.09 MPa, 35°C; Preferably, the ratio of water to methanol extract is 1:1 to 1:1.5 by mass. Preferably, the ratio of water to methanol extract is 1:1.5 by mass. Preferably, the ratio of ethyl acetate extraction to methanol extract complex solution is 1:1 to 1:4 by weight. Preferably, the ratio of ethyl acetate extraction to the weight ratio of the ethyl acetate to methanol extract complex solution is 1:

4. Preferably, the ethyl acetate extraction process is as follows: shaking for 0.5-1 h, standing for 1 h, and separating the ethyl acetate phase using a separator; Preferably, after the second ethyl acetate extraction, 1 mol / L hydrochloric acid is added to the aqueous phase solution to adjust the pH to 2.5-3.0, and then ethyl acetate solvent extraction is continued. Ethyl acetate is added at a mass ratio of 1:1 to 1:4 with the aqueous phase solution, shaken for 0.5-1 h, allowed to stand for 1 h, and the ethyl acetate phase is separated by a separator. The extraction is continued for 1-3 times. Preferably, after the second ethyl acetate extraction, 1 mol / L hydrochloric acid is added to the aqueous phase solution to adjust the pH to 3.0, and then ethyl acetate solvent extraction is continued. Ethyl acetate is added at a mass ratio of 1:4 with the aqueous phase solution, shaken for 0.5 h, allowed to stand for 1 h, and the ethyl acetate phase is separated by a separator. The extraction is then repeated twice. Preferably, the vacuum distillation conditions for the ethyl acetate extract are: vacuum degree below -0.09 MPa, 30℃~40℃; Preferably, the vacuum distillation conditions for the ethyl acetate extract are: vacuum degree below -0.09 MPa, 35°C; Preferably, the reconstitution ratio of the 50% methanol solution is 1:1 to 1:5 by mass of the 50% methanol solution to the ethyl acetate extract; Preferably, the reconstitution ratio of the 50% methanol solution is 1:2.5 (mass ratio of 50% methanol solution to ethyl acetate extract).

3. The purified component obtained by the preparation method according to claim 1 or 2.

4. A method for relieving pesticide damage, characterized in that, The method includes applying the purified and refined component as described in claim 3; Preferably, the pesticides include herbicides, insecticides, and fungicides; Preferably, the pesticide is nicosulfuron; Preferably, the pesticide is 2,4-D isooctyl ester.

5. The method according to claim 4, characterized in that, The method includes applying the purified and refined component of claim 3 prior to sowing; Preferably, the method of applying the purified and refined components is to soak the seeds after dilution; Preferably, the dilution factor is 10,000 to 50,000 times; Preferably, the dilution factor is 10,000 times; Preferably, the soaking temperature is 25~30℃; Preferably, the soaking time is 8-12 hours.

6. The method according to claim 4, characterized in that, The method includes applying the purified and refined component of claim 3 before applying pesticides; Preferably, the purified and refined components are applied by foliar spraying after dilution; Preferably, the dilution factor is 20,000 to 100,000 times; Preferably, the dilution factor is 20,000 times; Preferably, the purified and refined components are applied 2 to 3 times, with an interval of 3 to 5 days. Preferably, the method for applying the purified and refined components is as follows: before applying the pesticide, the purified and refined components are diluted and sprayed on the leaves, with an application interval of 3 to 5 days, and applied more than twice consecutively, and then the pesticide is applied. Preferably, the interval between the last application of the purified component and the application of the pesticide is within 48 hours.

7. The method according to claim 4, characterized in that, The method includes applying the purified and refined component of claim 3 after crop damage has occurred. Preferably, the purified and refined components are applied by foliar spraying after dilution. Preferably, the dilution factor is 20,000 to 100,000 times; Preferably, the dilution factor is 50,000 times; Preferably, the purified and refined components are applied 2 to 3 times, with an interval of 1 to 3 days. Preferably, the method for applying the purified and refined components is as follows: after the crop suffers phytotoxicity, the purified and refined components are diluted and sprayed on the stems and leaves, with an application interval of 1 to 3 days, and applied more than twice consecutively. Preferably, the purified and refined components are applied at the initial stage of drug-induced damage symptoms; Preferably, the purified and refined component is applied within 24 hours of the onset of phytotoxicity symptoms; Preferably, the purified and refined components are used to remove severely phytotoxic tissues from crops before application.

8. A method for enhancing the efficacy of pesticides, characterized in that, The method includes mixing and applying the purified and refined component of claim 3 with a pesticide; Preferably, the pesticides include herbicides, insecticides, and fungicides; Preferably, the pesticide is tebuconazole; Preferably, the method of mixed application is as follows: the purified and refined component is diluted, mixed with the pesticide, and applied to the soil surface and / or as a foliar spray; the amount of pesticide used is reduced by 10% to 20% compared to the method without the purified and refined component; Preferably, the dilution factor is 20,000 to 100,000 times; Preferably, the dilution factor is 20,000 times, and the amount of pesticide used is reduced by 20% compared to the unrefined component.

9. The application of the purified component according to claim 3, characterized in that, The application includes any of the following: 1) Application in eliminating pesticide residue damage in soil before sowing; 2) Application in relieving soil pesticide damage after sowing; 3) Application in preventing pesticide damage before applying pesticides to crops; 4) Application in promoting crop recovery after pesticide damage symptoms appear; 5) Application in enhancing pesticide efficacy and reducing pesticide usage.

10. The application according to claim 9, characterized in that, The pesticides include herbicides, insecticides, and fungicides; Preferably, the pesticide is nicosulfuron; Preferably, the pesticide is 2,4-D isooctyl ester; Preferably, the pesticide is tebuconazole; Preferably, the crop is selected from field crops and cash crops. Preferably, the crop is corn; Preferably, the crop is an apple.

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