A complex traditional Chinese medicine preparation for preventing and treating apple snails and a preparation method and application thereof

Abstract

The present application relates to the technical field of green prevention and control of rice diseases and pests, and specifically discloses a compound traditional Chinese medicine preparation for preventing and treating apple snails, and a preparation method and application thereof. The compound traditional Chinese medicine preparation comprises the following components in parts by weight: radix veratri 15-25 parts, monkshood 10-20 parts, Chinese toona fruit 15-20 parts, capsicum 18-28 parts, kopsia 8-15 parts, mint 5-10 parts, and rhizoma haemorrhoidalis 3-8 parts. The present application uses natural traditional Chinese medicine materials, has no chemical pesticide components, and can be completely degraded naturally without destroying the ecological balance of the rice field. The present application has the advantages of fast and complete killing speed, long egg inhibition duration, high overall killing rate of apple snails, wide source and low price of traditional Chinese medicine materials, simple preparation process, convenient field operation, multiple target points, no drug resistance of apple snails, continuous use for years, stable and non-declining prevention and control effect, improvement of the water and soil environment of the rice field, inhibition of the occurrence of diseases and pests in the field, promotion of the growth of rice roots, and improvement of the seedling emergence rate and seed setting rate of rice.

Description

Technical Field

[0001] This application relates to the field of green prevention and control technology for rice diseases and pests, specifically disclosing a compound traditional Chinese medicine preparation for controlling golden apple snails in rice, its preparation method, and its application. Background Technology

[0002] Golden apple snails are a major invasive alien pest in my country's rice-producing areas. They reproduce rapidly, feeding on rice seedlings and young panicles, seriously threatening rice yield and quality. Current control methods mainly rely on chemical agents and physical methods, both of which have significant limitations. Chemical control primarily uses metaldehyde and molluscicides, which are fast-acting but leave serious residues, damage paddy field aquatic ecosystems and biodiversity, and induce drug resistance in golden apple snails with long-term use, leading to diminishing control effectiveness and a vicious cycle of drug use. Furthermore, the high risk of pesticide residues in rice makes it unsuitable for green and organic rice standards. Physical control methods, such as manual collection, water network interception, and duck farming in rice paddies, are environmentally friendly, but manual collection is inefficient and costly, suitable only for small areas; water network interception cannot eradicate existing snail populations; and duck farming for snail control is affected by weather and the snail-growing period, resulting in poor stability and making large-scale implementation difficult. Existing plant-based snail control technologies also have shortcomings such as single formula, low snail-killing rate, short effective period, susceptibility to environmental influences, and risk of seedling damage. There is a lack of mature alternatives suitable for large-scale planting.

[0003] Current technologies for controlling golden apple snails in rice paddies have the following drawbacks: First, chemical molluscicides leave high residues, kill non-target beneficial organisms, and damage biodiversity in paddy fields, posing ecological safety risks. Second, long-term use of single-drug therapy leads to the evolution of drug resistance in golden apple snails and a decline in control efficacy, making it difficult to overcome resistance bottlenecks and achieve stable and long-term eradication. Third, physical and biological control methods are inefficient, highly dependent on environmental conditions, and difficult to standardize and promote, hindering the development of replicable solutions suitable for large-scale contiguous planting. Fourth, existing plant-derived formulations have low molluscicidal rates, short durations of action, high environmental sensitivity, and potential seedling damage, requiring optimization of formulation systems to balance efficacy, safety, and crop compatibility. Summary of the Invention

[0004] In view of this, this application provides a compound traditional Chinese medicine preparation for the control of golden apple snails in rice, its preparation method and application, in order to solve the technical problems of chemical pesticide residue pollution, strong pesticide resistance of pests, damage to the ecological environment, low efficiency and high cost of physical control, inability to be promoted on a large scale, and poor effect and insufficient stability of conventional plant-derived pesticides in the existing control technologies for golden apple snails in rice.

[0005] In a first aspect, the present invention provides a compound traditional Chinese medicine preparation for the prevention and control of golden apple snails in rice, comprising the following parts by weight: 15-25 parts of Veratrum nigrum, 10-20 parts of Aconitum carmichaelii, 15-20 parts of Melia toosendan, 18-28 parts of chili pepper, 8-15 parts of Citrus reticulata, 5-10 parts of Mentha haplocalyx, and 3-8 parts of Acorus tatarinowii.

[0006] Furthermore, the preferred weight parts of the compound traditional Chinese medicine preparation for controlling golden apple snails in rice according to the present invention are: 16-24 parts of Veratrum nigrum, 11-19 parts of Aconitum carmichaelii, 16-19 parts of Melia toosendan, 19-27 parts of chili pepper, 9-14 parts of Citrus reticulata, 6-9 parts of Mentha haplocalyx, and 4-7 parts of Acorus tatarinowii.

[0007] Furthermore, the optimal weight proportions of the compound traditional Chinese medicine preparation for controlling golden apple snails in rice according to the present invention are: 20 parts of Veratrum nigrum, 10 parts of Aconitum carmichaelii, 20 parts of Melia toosendan, 20 parts of chili pepper, 15 parts of Citrus reticulata, 8 parts of Mentha haplocalyx, and 7 parts of Acorus tatarinowii.

[0008] Secondly, the present invention also provides a method for preparing a compound traditional Chinese medicine preparation for controlling golden apple snails in rice, comprising the following steps: S1. Place the raw materials of Veratrum nigrum, Aconitum carmichaelii, Melia toosendan, chili pepper, bitter vine, mint and Acorus tatarinowii in a ventilated and shady place to air dry naturally, then crush them and pass them through a 40-60 mesh sieve to obtain mixed Chinese medicine powder. S2. Add water to the mixed Chinese herbal powder at a mass-volume ratio of 1:8 to 1:10, add 10% of the mass of tea seed cake or cake residue of the mixed Chinese herbal powder, and then add fermentation bacteria composed of Bacillus subtilis, Lactobacillus plantarum and compound enzymes. The inoculation amount of the fermentation bacteria is 5% to 8% of the total mass of the mixed powder and tea seed cake or cake residue to obtain Chinese herbal fermentation liquid. S3. First, the fermented Chinese medicine liquid is coarsely filtered through a 60-80 mesh filter. Then, a 0.45μm cellulose acetate filter membrane is used to perform vacuum negative pressure fine filtration on the coarsely filtered fermented Chinese medicine liquid. The residue is then removed and the filtrate is collected. The filtrate is then concentrated at normal pressure and low temperature to reduce the volume of the filtrate to 1 / 3 of the original volume of the fermented Chinese medicine liquid, thus obtaining the concentrated mother liquor of fermented Chinese medicine. S4. Add natural plant adjuvants according to the conventional adjuvant addition amount to the concentrated mother liquor of fermented Chinese medicine, stir evenly and let stand to defoam, and you will get a compound Chinese medicine preparation for the control of golden apple snail in rice that can be applied directly. Store in a sealed container away from light for later use.

[0009] Preferably, in S2, the fermentation temperature of the obtained traditional Chinese medicine fermentation liquid is 30℃~35℃, the fermentation time is 7~10 days, and the fermentation is fully carried out. The bacterial enzymes break the cell walls, and the large molecules in the traditional Chinese medicine are transformed into small molecules through fermentation and enzymatic hydrolysis, so that the effective components in the traditional Chinese medicine raw materials are fully released.

[0010] Preferably, in step S2, the pH value of the traditional Chinese medicine fermentation broth is 4.5~5.0, and the viable count is ≥2×10⁻⁶. 9 CFU / mL.

[0011] Preferably, in S3, the conditions for performing vacuum negative pressure fine filtration are: temperature 30℃~35℃, vacuum degree 0.06MPa~0.07MPa, and feed rate 8~10L / h.

[0012] Preferably, in S3, the transmittance of the filtrate after vacuum negative pressure filtration is ≥98%, and the retention rate of effective components is ≥95%.

[0013] Thirdly, the present invention also provides the application of the compound traditional Chinese medicine preparation in the control of golden apple snails in rice paddies, by applying the compound traditional Chinese medicine preparation to rice fields.

[0014] Preferably, the application includes the following steps: S100. One to two days before applying the pesticide, prepare the paddy field ditches and maintain a shallow water layer in the paddy field. The water depth should be controlled at 3cm to 5cm to ensure that the water surface of the paddy field evenly covers the soil surface, with no waterlogged dead corners or dry and cracked plots. S200. Dilute the prepared compound Chinese medicine preparation with water at a ratio of 1:150 to 1:200 and stir evenly. Quantitatively prepare the medicine according to the area of ​​the paddy field, using 30 kg to 40 kg of the medicine solution per mu of paddy field. S300. On a sunny day, between 9:00 and 11:00 in the morning or between 16:00 and 18:00 in the afternoon, use a backpack sprayer or drone to evenly spray the diluted compound Chinese medicine preparation on the entire paddy field. The water depth for spraying should be 3cm to 5cm. Focus on spraying the base of rice seedlings, the edge of the field ridges, and the areas around ditches where golden apple snails gather. S400: Seven days after the first application, a second application is made to kill any remaining adult snails and newly hatched juvenile snails. A third preventative application is made at the early tillering stage of rice to achieve snail-free growth throughout the entire growth period. S500: Keep the shallow water layer in the paddy field without draining or changing the water for 3 days after application to ensure that the compound Chinese medicine preparation can fully exert its efficacy. Normal field water and fertilizer management can be carried out after 3 days.

[0015] After adopting the above technical solution, the beneficial effects of the present invention are as follows: (1) The whole process uses natural Chinese medicinal materials, without chemical pesticides. The agent can be completely biodegraded naturally, with no residues in soil, water, or rice. It does not damage the ecological balance of paddy fields, protects the biodiversity of paddy fields, and meets the development standards of green agriculture and organic agriculture. (2) The four-fold synergistic effect can kill snails quickly and thoroughly, inhibit eggs for a long time, and achieve an overall kill rate of over 95%, which is far superior to conventional physical control and ordinary plant-based control technologies. It can completely solve the problem of snail damage in rice fields. (3) Chinese medicinal materials are widely available and inexpensive, with simple preparation processes and convenient field operations. They do not require high equipment investment and labor costs. Compared with long-term chemical application and manual picking and control, the overall control cost is reduced by more than 40%. (4) The compound effect of multiple components of natural plants has multiple targets, and the golden apple snail cannot develop drug resistance. It can be used continuously for many years, and the control effect is stable and does not decrease. (5) The active ingredients of the agent can improve the soil and water environment of paddy fields, inhibit the occurrence of pests and diseases in the field, promote the growth of rice roots, and increase the rice seedling rate and grain filling rate. While preventing pests, it can also increase rice yield and improve quality, resulting in significant economic, ecological and social benefits. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 This is the process flow of the compound traditional Chinese medicine preparation for the control of golden apple snail in rice, as described in this application. Figure 1 ; Figure 2 This is the process flow of the compound traditional Chinese medicine preparation for the control of golden apple snail in rice, as described in this application. Figure 2 ; Figure 3 This is a flowchart illustrating the application of the compound traditional Chinese medicine preparation of this application in the control of golden apple snails in rice. Detailed Implementation

[0018] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application. In addition, it should be understood that the specific implementation methods described herein are only for illustration and explanation of this application and are not intended to limit this application.

[0019] In the description of this application, the term "comprising" means "including but not limited to". The terms first, second, third, etc. are used merely as illustrative purposes and do not impose numerical requirements or establish an order.

[0020] In this application, "and / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A existing alone, A and B existing simultaneously, or B existing alone. A and B can be singular or plural.

[0021] In this application, "at least one" means one or more, and "more than one" means two or more. "One or more", "at least one of the following", or similar expressions refer to any combination of these items, including any combination of single or multiple items. For example, "at least one of a, b, or c", or "at least one of a, b, and c", can both mean: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can be single or multiple.

[0022] Various embodiments of this application may exist in the form of a range; it should be understood that the description in the form of a range is merely for convenience and brevity and should not be construed as a rigid limitation on the scope of this application; therefore, it should be considered that the range description has specifically disclosed all possible sub-ranges and single numerical values ​​within that range. For example, it should be considered that the range description from 1 to 6 has specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and single numbers within the range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. Furthermore, whenever a numerical range is referred to herein, it means including any referenced number (fraction or integer) within the referred range.

[0023] The present application will be specifically described below through specific embodiments. The following embodiments are only some embodiments of the present application and are not intended to limit the present application.

[0024] Example 1 This invention provides a compound traditional Chinese medicine preparation for the prevention and control of golden apple snails in rice, comprising the following parts by weight: 20 parts of Veratrum nigrum, 10 parts of Aconitum carmichaelii, 20 parts of Melia toosendan, 20 parts of Citrus reticulata, 15 parts of Citrus reticulata bark, 7 parts of Mentha haplocalyx, and 8 parts of Acorus tatarinowii.

[0025] Specifically, the active substances in Veratrum nigrum that kill snails are mainly steroidal alkaloids, with core components including veratrine, proveratrine, pseudoveratrine, red veratrine, veratramine, and veratrin. It has extremely strong contact and stomach poison effects, can quickly destroy the nervous system of golden apple snails, achieve rapid killing, and also has the effects of inhibiting golden apple snail eggs and repelling them.

[0026] The molluscicidal active substances in *Melia azedarach* are mainly limonene triterpenoids, including: azadirachtin, azadirachtinone, azadirachtin alcohol, and azadirachtin aldehyde, with contents of 0.06%–0.20%. It exhibits five activities against *Pomacea canaliculata* (rice golden apple snail): antifeedant, stomach poison, contact toxicity, oviposition inhibition, and repellency, and is highly effective against mollusks.

[0027] The main active ingredient in chili peppers for killing molluscs is capsaicin, accounting for 70%–90% of the total alkaloids in chili peppers; the minor components are dihydrocapsaicin, nordihydrocapsaicin, homocapsaicin, etc., collectively known as capsaicinoids. It has strong irritant, neurotoxic, stomach poison, repellent, and antifeedant effects on rice golden apple snails, and is highly toxic to both juvenile and adult snails of rice golden apple snails.

[0028] The molluscicidal active substances of *Cephalotaxus fortunei* are mainly dihydrolinuron sesquiterpene esters (e.g., cephalotaxin), including: cephalotaxin V, the main toxic agent and the most potent stomach toxicity; cephalotaxin IV, which has anesthetic activity and causes muscle relaxation; and cephalotaxin I–III, angulatin A, etc., which have synergistic effects of anorexicism and oocyte inhibition. *Cephalotaxus fortunei* primarily exerts its stomach toxicity, with contact toxicity as a secondary effect, providing strong anorexicism and anesthesia, making it highly effective and with low residual effects on mollusks.

[0029] The molluscicidal or molluscicidal activity of peppermint is mainly due to its volatile oils (e.g., peppermint oil). Key components include: menthol (60%–70%), which has a strong repellent and neuro-anesthetic effect on *Pomacea canaliculata* (rice golden apple snail); menthone and pulegone, which produce stomach toxins and inhibit oviposition in *Pomacea canaliculata*; and 1,8-cineole, limonene, and α-pinene contained in peppermint, which synergistically enhance its effects. Peppermint primarily acts as a repellent, with secondary stomach toxin effects, strong oviposition inhibition, and low toxicity and safety. Adult snails show significant repulsion, while juvenile snails are sensitive, and it has an inhibitory effect on eggs.

[0030] The molluscicidal or molluscicidal activity of Acorus tatarinowii is mainly due to its volatile oil, with core components including: strong repellent, neurotoxic, and gastric toxins such as α-asarone and β-asarone (e.g., accounting for 70%–80% of the volatile oil); synergistic anesthesia, ovicidal inhibition, and anorexic effects of acorylether, acorylol, and terpenes. It exhibits strong repellency against golden apple snails in rice, moderate gastric toxicity, significant ovicidal inhibition, and low residue safety; it is particularly effective against adult snails, sensitive to juvenile snails, and has a high inhibition rate on egg mass hatching; the naturally occurring volatile oil can modulate the efficacy of the pesticide, reduce the irritation of raw materials, and simultaneously improve the diffusion and persistence of the pesticide, avoiding phytotoxicity to rice seedlings.

[0031] Aconite contains C as its key activity. 19 - Diterpenoid diester alkaloids, with three core components: aconitine, the most toxic and potent molluscicide; mesonaconitine, highly active and rapidly penetrating; and hypoaconitine, abundant and long-lasting. Aconitum contains approximately 0.8%–1.2% total alkaloids, possessing extremely strong contact and stomach poison effects. It can paralyze the muscle system of the golden apple snail, inhibiting its activity, feeding, and reproduction.

[0032] This invention also provides a method for preparing a compound traditional Chinese medicine preparation for controlling golden apple snails in rice, comprising the following steps: S1. Place the raw materials of Veratrum nigrum, Aconitum carmichaelii, Melia toosendan, chili pepper, bitter vine, mint and Acorus tatarinowii in a ventilated and shady place to air dry naturally, then crush them and pass them through a 60-mesh sieve to obtain mixed Chinese medicine powder. S2. Add water to the mixed Chinese herbal powder at a mass-to-volume ratio of 1:10, add 10% (by weight) of tea seed cake or oilseed cake from the mixed Chinese herbal powder, and then add a fermentation bacteria composed of Bacillus subtilis, Lactobacillus plantarum, and a compound enzyme. The inoculum amount of the fermentation bacteria is 8% of the total mass of the mixed powder and tea seed cake or oilseed cake. The fermentation temperature is 35℃, and the fermentation time is 10 days. Fermentation is allowed to proceed fully, the bacterial and enzyme cells are broken down, and the large molecules in the Chinese herbal medicine are converted into smaller molecules through fermentation and enzymatic hydrolysis, allowing the effective components in the raw materials to be fully released. The pH of the fermentation liquid should be 5.0, and the viable cell count should be ≥2×10⁻⁶. 9 CFU / mL was used to obtain the fermentation broth of traditional Chinese medicine; S3. First, the fermentation liquid of traditional Chinese medicine is coarsely filtered through an 80-mesh filter. Then, under the conditions of 35℃, vacuum degree 0.07MPa, and feed rate 10L / h, it is finely filtered under vacuum negative pressure using a 0.45μm cellulose acetate filter membrane. The transmittance of the filtrate is ≥98%, and the retention rate of effective components is ≥95%. Then, the filtrate is collected after removing the dregs. The filtrate is then concentrated at normal pressure and low temperature to reduce the volume of the filtrate to 1 / 3 of the original volume of the fermentation liquid of traditional Chinese medicine, thus obtaining the concentrated mother liquor of fermented traditional Chinese medicine. S4. Add natural plant adjuvants according to the conventional adjuvant addition amount to the concentrated mother liquor of fermented Chinese medicine, stir evenly and let stand to defoam, and you will get a compound Chinese medicine preparation for the control of golden apple snail in rice that can be applied directly. Store in a sealed container away from light for later use.

[0033] This invention also provides the application of the above-mentioned compound traditional Chinese medicine preparation in the control of golden apple snails in rice, the application including the following steps: S100. One day before applying the pesticide, prepare the paddy field ditches and maintain a shallow water layer in the paddy field, with the water depth controlled at 3cm. Ensure that the water surface of the paddy field evenly covers the soil surface, with no waterlogged dead corners or dry and cracked plots. S200. Dilute the prepared compound Chinese medicine preparation with water at a ratio of 1:150 and stir evenly. Quantitatively prepare the medicine according to the area of ​​the paddy field, using 35 kg of liquid medicine per mu of paddy field. S300. On a sunny morning at 10:00, use a backpack sprayer or drone to evenly spray the diluted compound Chinese medicine preparation on the entire paddy field. The water depth for spraying should be 3cm. Focus on spraying the base of rice seedlings, the edge of the field ridges, and the areas around ditches where golden apple snails gather. S400: Seven days after the first application, a second application is made to kill any remaining adult snails and newly hatched juvenile snails. A third preventative application is made at the early tillering stage of rice to achieve snail-free growth throughout the entire growth period. S500: Keep the shallow water layer in the paddy field without draining or changing the water for 3 days after application to ensure that the compound Chinese medicine preparation can fully exert its efficacy. Normal field water and fertilizer management can be carried out after 3 days.

[0034] As a result, after 3 days of application, the mortality rate of golden apple snails in the field reached 95%; after 10 days of application, there were very few remaining juvenile snails and newly hatched snails, and the overall eradication rate reached 96.2%; there was no golden apple snail damage throughout the entire growth period, the rice seedlings grew vigorously, the paddy field water was clear, and there were no pesticide damage or pollution phenomena.

[0035] Example 2 This invention provides a compound traditional Chinese medicine preparation for the prevention and control of golden apple snails in rice, comprising the following parts by weight: 15 parts of Veratrum nigrum, 15 parts of Aconitum carmichaelii, 15 parts of Melia toosendan, 25 parts of Citrus reticulata, 15 parts of Citrus reticulata bark, 8 parts of Mentha haplocalyx, and 7 parts of Acorus tatarinowii.

[0036] This invention also provides a method for preparing a compound traditional Chinese medicine preparation for controlling golden apple snails in rice, comprising the following steps: S1. Place the raw materials of Veratrum nigrum, Aconitum carmichaelii, Melia toosendan, chili pepper, bitter vine, mint and Acorus tatarinowii in a ventilated and shady place to air dry naturally, then crush them and pass them through a 60-mesh sieve to obtain mixed Chinese medicine powder. S2. Add water to the mixed Chinese herbal powder at a mass-to-volume ratio of 1:8, add 10% (by weight) of tea seed cake or oilseed cake from the mixed Chinese herbal powder, and then add a fermentation bacteria composed of Bacillus subtilis, Lactobacillus plantarum, and a compound enzyme. The inoculum amount of the fermentation bacteria is 8% of the total mass of the mixed powder and tea seed cake or oilseed cake. The fermentation temperature is 30℃, and the fermentation time is 8 days. Fermentation is allowed to proceed fully, the bacterial and enzyme cells are broken down, and the large molecules in the Chinese herbal medicine are converted into smaller molecules through fermentation and enzymatic hydrolysis, allowing the effective components in the raw materials to be fully released. The pH value of the fermentation liquid is 5.0, and the viable cell count is ≥2×10⁻⁶. 9 CFU / mL was used to obtain the fermentation broth of traditional Chinese medicine; S3. First, the fermentation liquid of traditional Chinese medicine is coarsely filtered through an 80-mesh filter. Then, under the conditions of 35℃, vacuum degree 0.07MPa, and feed rate 10L / h, it is finely filtered under vacuum negative pressure using a 0.45μm cellulose acetate filter membrane. The transmittance of the filtrate is ≥98%, and the retention rate of effective components is ≥95%. Then, the filtrate is collected after removing the dregs. The filtrate is then concentrated at normal pressure and low temperature to reduce the volume of the filtrate to 1 / 3 of the original volume of the fermentation liquid of traditional Chinese medicine, thus obtaining the concentrated mother liquor of fermented traditional Chinese medicine. S4. Add natural plant adjuvants according to the conventional adjuvant addition amount to the concentrated mother liquor of fermented Chinese medicine, stir evenly and let stand to defoam, and you will get a compound Chinese medicine preparation for the control of golden apple snail in rice that can be applied directly. Store in a sealed container away from light for later use.

[0037] This invention also provides the application of the above-mentioned compound traditional Chinese medicine preparation in the control of golden apple snails in rice, the application including the following steps: S100. One day before applying the pesticide, prepare the paddy field ditches and maintain a shallow water layer in the paddy field, with the water depth controlled at 5cm. Ensure that the water surface of the paddy field evenly covers the soil surface, with no waterlogged dead corners or dry and cracked plots. S200. Dilute the prepared compound Chinese medicine preparation with water at a ratio of 1:200 and stir evenly. Quantitatively prepare the medicine according to the area of ​​the paddy field, using 40 kg of medicine solution per mu of paddy field. S300. On a sunny morning at 10:00, use a backpack sprayer or drone to evenly spray the diluted compound Chinese medicine preparation on the entire paddy field. The water depth for spraying should be 3cm. Focus on spraying the base of rice seedlings, the edge of the field ridges, and the areas around ditches where golden apple snails gather. S400: Seven days after the first application, a second application is made to kill any remaining adult snails and newly hatched juvenile snails. A third preventative application is made at the early tillering stage of rice to achieve snail-free growth throughout the entire growth period. S500: Keep the shallow water layer in the paddy field without draining or changing the water for 3 days after application to ensure that the compound Chinese medicine preparation can fully exert its efficacy. Normal field water and fertilizer management can be carried out after 3 days.

[0038] Therefore, it can effectively prevent golden apple snails from invading rice paddies, inhibiting snail egg hatching by more than 95%, leaving rice paddies basically free of snail damage and having no adverse effects on seedlings.

[0039] Example 3 This invention provides a compound traditional Chinese medicine preparation for the prevention and control of golden apple snails in rice, comprising the following parts by weight: 20 parts of Veratrum nigrum, 10 parts of Aconitum carmichaelii, 20 parts of Melia toosendan, 20 parts of Citrus reticulata, 15 parts of Citrus reticulata bark, 7 parts of Mentha haplocalyx, and 8 parts of Acorus tatarinowii.

[0040] This invention also provides a method for preparing a compound traditional Chinese medicine preparation for controlling golden apple snails in rice, comprising the following steps: S1. Place the raw materials of Veratrum nigrum, Aconitum carmichaelii, Melia toosendan, chili pepper, bitter vine, mint and Acorus tatarinowii in a ventilated and shady place to air dry naturally, then crush them and pass them through a 60-mesh sieve to obtain mixed Chinese medicine powder. S2. Add water to the mixed Chinese herbal powder at a mass-to-volume ratio of 1:8, add 10% (by weight) of tea seed cake or oilseed cake from the mixed Chinese herbal powder, and then add a fermentation bacteria composed of Bacillus subtilis, Lactobacillus plantarum, and a compound enzyme. The inoculum amount of the fermentation bacteria is 8% of the total mass of the mixed powder and tea seed cake or oilseed cake. The fermentation temperature is 35℃, and the fermentation time is 7 days. Fermentation is complete, and the bacterial and enzyme cell walls are broken down, converting the large molecules in the Chinese herbal medicine into smaller molecules through fermentation and enzymatic hydrolysis, thus fully releasing the effective components in the raw materials. The pH of the fermentation liquid should be 5.0, and the viable cell count should be ≥2×10⁻⁶. 9 CFU / mL was used to obtain the fermentation broth of traditional Chinese medicine; S3. First, the fermentation liquid of traditional Chinese medicine is coarsely filtered through an 80-mesh filter. Then, under the conditions of 35℃, vacuum degree 0.07MPa, and feed rate 10L / h, it is finely filtered under vacuum negative pressure using a 0.45μm cellulose acetate filter membrane. The transmittance of the filtrate is ≥98%, and the retention rate of effective components is ≥95%. Then, the filtrate is collected after removing the dregs. The filtrate is then concentrated at normal pressure and low temperature to reduce the volume of the filtrate to 1 / 3 of the original volume of the fermentation liquid of traditional Chinese medicine, thus obtaining the concentrated mother liquor of fermented traditional Chinese medicine. S4. Add natural plant adjuvants according to the conventional adjuvant addition amount to the concentrated mother liquor of fermented Chinese medicine, stir evenly and let stand to defoam, and you will get a compound Chinese medicine preparation for the control of golden apple snail in rice that can be applied directly. Store in a sealed container away from light for later use.

[0041] This invention also provides the application of the above-mentioned compound traditional Chinese medicine preparation in the control of golden apple snails in rice, the application including the following steps: S100. One day before applying the pesticide, prepare the paddy field ditches and maintain a shallow water layer in the paddy field, with the water depth controlled at 4cm. Ensure that the water surface of the paddy field evenly covers the soil surface, with no waterlogged dead corners or dry and cracked plots. S200. Dilute the prepared compound Chinese medicine preparation with water at a ratio of 1:180 and stir evenly. Quantitatively prepare the medicine according to the area of ​​the paddy field, using 40 kg of medicine solution per mu of paddy field. S300. On a sunny morning at 10:00, use a backpack sprayer or drone to evenly spray the diluted compound Chinese medicine preparation on the entire paddy field. The water depth for spraying should be 4cm. Focus on spraying the base of rice seedlings, the edge of the field ridges, and the areas around ditches where golden apple snails gather. S400: Seven days after the first application, a second application is made to kill any remaining adult snails and newly hatched juvenile snails. A third preventative application is made at the early tillering stage of rice to achieve snail-free growth throughout the entire growth period. S500: Keep the shallow water layer in the paddy field without draining or changing the water for 3 days after application to ensure that the compound Chinese medicine preparation can fully exert its efficacy. Normal field water and fertilizer management can be carried out after 3 days.

[0042] Therefore, the treatment is effective within 2 days, causing adult snails to die quickly, with an overall eradication rate of 95.8% and a duration of effectiveness of up to 20 days, effectively controlling the secondary outbreak of golden apple snails.

[0043] It should be noted that Aconitum carmichaelii fermentation is a highly efficient and green technology for reducing toxicity and enhancing efficacy. It can reduce highly toxic diester alkaloids to a safe range while retaining their medicinal effects. Its safety is significantly better than that of raw Aconitum carmichaelii and some traditionally processed products. Fermentation can significantly reduce the toxicity of Aconitum carmichaelii, and its safety is controllable. The toxicity reduction mechanism is clear, and the medicinal effects are well preserved, but strict control of the strains and processes is required.

[0044] Specifically, the toxic source of Aconitum carmichaelii is diester alkaloids (DDAs) – aconitine, neoaconitine, and hypoaconitine. The detoxification mechanism may include esterases produced by microorganisms (e.g., yeast, mold, probiotics, etc.) to hydrolyze highly toxic diester alkaloids (DDAs) into less toxic monoester alkaloids (MDAs) and non-toxic alcohol amine alkaloids (ADAs), reducing toxicity by 64–180 times while retaining cardiotonic and analgesic activities.

[0045] Among them, raw Aconitum carmichaelii contains approximately 0.19% diester alkaloids (DDAs) and approximately 0.005% low-toxicity monoester alkaloids (MDAs); after fermentation, the content of diester alkaloids (DDAs) drops to below 0.001%, while the content of low-toxicity monoester alkaloids (MDAs) rises to 0.01%–0.03%.

[0046] Aconite Root Toxicity Reduction Mechanism Verification Experiment The mechanism of aconite toxicity reduction can be verified through in vitro, animal, and quality control experiments. First, regarding in vitro toxicity, the fermented extract of Aconitum carmichaelii has an effect on the sodium content of cardiomyocytes. + Channel inhibition was significantly reduced, and the risk of arrhythmia was decreased; hemolysis and cytotoxicity tests showed a 5–10-fold increase in safe concentration.

[0047] Secondly, acute toxicity (LD50) in animals 50 A gavage test was conducted on mice using raw aconite, and the result was LD50. 50 The LD50 concentration in raw aconite is approximately 3.5 g / kg, at which point it is considered highly toxic. An oral gavage test on mice using fermented aconite showed an LD50 of approximately 3.5 g / kg. 50 At concentrations >30g / kg, fermented aconite is in a low-toxicity or non-toxic state, with no deaths or significant liver and kidney damage. Long-term toxicity was observed for 90 days, and no abnormalities were found in liver and kidney function, blood routine tests, or organ pathology, with no cumulative toxicity.

[0048] Finally, for quality control, the total amount of diester alkaloids is ≤0.0015% (e.g., fermented products are mostly <0.0006%, far below the limit), and the amount of monoester alkaloids is ≥0.010% (e.g., fermented products are stable at 0.01%–0.03%).

[0049] Therefore, fermented aconite can retain its core medicinal effects, and its cardiotonic, anti-heart failure, anti-inflammatory, analgesic, and immunomodulatory effects are comparable to or stronger than those of traditionally processed aconite. The fermentation conditions are mild (e.g., fermentation temperature 30℃, fermentation time 7 days), with low energy consumption, short cycle, and controllable composition, which can avoid the loss of effective components caused by traditional steaming and boiling.

[0050] Drug efficacy verification test To verify the killing effect of the compound traditional Chinese medicine preparation of this invention on golden apple snails, the following experiments were conducted: To verify the killing activity of the compound traditional Chinese medicine preparation of this invention against *Pomacea canaliculata* in rice, an indoor toxicity test was conducted, as follows: Experimental subjects: adult golden apple snails (e.g., shell height 2.5~3.5cm) and juvenile snails (e.g., shell height 0.8~1.5cm) collected from rice fields. Before the experiment, they were acclimatized for 3 days under room temperature (e.g., 25±2℃) and light or darkness conditions for 12 hours. Individuals with consistent vitality were selected for the experiment.

[0051] Experimental Methods: The immersion method was used. The compound traditional Chinese medicine preparation prepared in Example 1 was diluted with distilled water to create a series of concentration gradients (e.g., 1.0 g / L, 2.0 g / L, 3.0 g / L, 4.0 g / L, 5.0 g / L, 6.0 g / L). Three replicates were set for each concentration, with 20 snails per replicate. Water was used as a blank control. Mortality was observed and recorded at 12 h, 24 h, 48 h, and 72 h after treatment. Snails without a reaction when their legs were lightly touched with a brush were considered dead. The median lethal concentration (LC50) was calculated using SPSS software. 50 (and 95% confidence interval).

[0052] Experimental results: The LC50 of the compound traditional Chinese medicine preparation against juvenile golden apple snails for 72 hours was... 50 The value was 3.0 g / L, and the LC value of the 72-hour mollusc was 3.0 g / L. 50 The concentration was 5.5 g / L, indicating that the formulation was more sensitive to juvenile snails and had stronger killing activity. The time effect results showed that under the condition of formulation concentration of 6.0 g / L, the mortality rate of juvenile snails was 56% after 12 hours, 95.5% after 24 hours, 99.8% after 48 hours, and 100% after 72 hours, indicating that the formulation has good rapid effect and can achieve near-complete eradication within 24 hours after application.

[0053] To further verify the synergistic effect of the compound traditional Chinese medicine preparation of the present invention with other plant-derived molluscicides, matrine (e.g., purity ≥90%) was selected and combined with the compound traditional Chinese medicine preparation of the present invention for toxicity determination, as follows: The experimental method was the same as the indoor toxicity assay, with three treatment groups: ① single-dose compound traditional Chinese medicine preparation group; ② single-dose matrine group; ③ compound traditional Chinese medicine preparation and matrine mixed at a mass ratio of 5:1, forming a compound group. The LC50 of each group was calculated at 24 h. 50 The values ​​were calculated, and the co-toxicity coefficient (CTC) was calculated according to Sun Yunpei's method. A CTC ≥ 120 was considered synergistic, a CTC ≥ 80 and < 120 was considered additive, and a CTC < 80 was considered antagonistic.

[0054] Experimental results: LC50 of single-dose compound traditional Chinese medicine preparations over 24 hours 50 The value was 5.5 g / L; the LC50 of a single dose of matrine over 24 hours was... 50 The value was 7.2 g / L; the LC50 of the compound group after 24 hours was... 50The value was 3.8 g / L, and the co-toxicity coefficient CTC=220, which is much greater than 120. This indicates that the compound traditional Chinese medicine preparation and matrine produced a strong synergistic effect after being compounded at a mass ratio of 5:1, and the molluscicidal activity was significantly improved.

[0055] To verify the practical application effect of the compound traditional Chinese medicine preparation of this invention, a field efficacy test was conducted in a rice field with a severe infestation of golden apple snails, 7 days after rice transplanting (e.g., early tillering stage). The test site was located in a typical rice-growing area in South China. The soil in the test field was clay loam with a pH of 6.2–6.8, and the average population of golden apple snails was 15–20 per m². 2 .

[0056] The experiment consisted of two treatment groups: ① Experimental group: 25 kg of the compound traditional Chinese medicine preparation prepared in Example 1 of this invention was applied per mu (approximately 0.067 hectares), mixed with 5 kg of 0.3% matrine aqueous solution, diluted at a ratio of 1:150 and sprayed evenly; ② Control group: 10 kg of 0.3% matrine aqueous solution per mu (approximately 0.067 hectares), diluted at a ratio of 1:150 and sprayed. Each group had three replicates, with a plot area of ​​100 m². 2 Randomized block design. Application conditions: Spray at 10:00 AM on a sunny day, with a water depth of 3-5 cm. Do not drain the water for 3 days after application.

[0057] The control efficacy was investigated 7 days after application. Control efficacy (%) = (number of live snails in the control group - number of live snails in the treatment group) / number of live snails in the control group × 100%.

[0058] The experimental results showed that after 7 days of application, the control efficacy of the experimental group for golden apple snails reached 96.4%, with very few live snails remaining in the field and no gaps or breaks in the rice seedling rows. In contrast, the control group, using matrine alone, had a control efficacy of only 65.7%, showing a significant difference. This indicates that the compound traditional Chinese medicine preparation of this invention, whether used alone or in combination with other plant-derived molluscicides, has excellent field control effects. In particular, the synergistic effect is significant when combined with matrine, enabling efficient and green control of golden apple snails.

[0059] The above are all preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Therefore, all equivalent modifications, equivalent improvements or equivalent substitutions made based on the formula, process, structure and principle of the present invention should be included within the scope of protection of the present invention.

Claims

1. A compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice, characterized in that, It includes the following parts by weight: 15-25 parts of Veratrum nigrum, 10-20 parts of Aconitum carmichaelii, 15-20 parts of Melia toosendan, 18-28 parts of chili pepper, 8-15 parts of Bitter Orange Vine, 5-10 parts of Peppermint, and 3-8 parts of Acorus calamus.

2. The compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice according to claim 1, characterized in that, It includes the following parts by weight: 16-24 parts of Veratrum nigrum, 11-19 parts of Aconitum carmichaelii, 16-19 parts of Melia toosendan, 19-27 parts of chili pepper, 9-14 parts of Citrus reticulata, 6-9 parts of Mentha haplocalyx, and 4-7 parts of Acorus calamus.

3. The compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice according to claim 1, characterized in that, The ingredients include the following parts by weight: 20 parts of Veratrum nigrum, 10 parts of Aconitum carmichaelii, 20 parts of Melia toosendan, 20 parts of chili pepper, 15 parts of Bitter Orange Vine, 8 parts of Peppermint, and 7 parts of Acorus calamus.

4. A method for preparing a compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice, as described in any one of claims 1-3, characterized in that... Includes the following steps: S1. Place the raw materials of Veratrum nigrum, Aconitum carmichaelii, Melia toosendan, chili pepper, bitter vine, mint and Acorus tatarinowii in a ventilated and cool place to air dry naturally, then crush them and pass them through a 40-60 mesh sieve to obtain mixed Chinese medicine powder. S2. Add water to the mixed Chinese herbal powder at a mass-volume ratio of 1:8 to 1:10, add 10% of the mass of tea seed cake or cake residue of the mixed Chinese herbal powder, and then add fermentation bacteria composed of Bacillus subtilis, Lactobacillus plantarum and compound enzymes. The inoculation amount of the fermentation bacteria is 5% to 8% of the total mass of the mixed powder and tea seed cake or cake residue to obtain Chinese herbal fermentation liquid. S3. First, the fermented Chinese medicine liquid is coarsely filtered through a 60-80 mesh filter. Then, a 0.45μm cellulose acetate filter membrane is used to perform vacuum negative pressure fine filtration on the coarsely filtered fermented Chinese medicine liquid. The residue is then removed and the filtrate is collected. The filtrate is then concentrated at normal pressure and low temperature to reduce the volume of the filtrate to 1 / 3 of the original volume of the fermented Chinese medicine liquid, thus obtaining the concentrated mother liquor of fermented Chinese medicine. S4. Add natural plant adjuvants according to the conventional adjuvant addition amount to the concentrated mother liquor of fermented Chinese medicine, stir evenly and let stand to defoam, and you will get a compound Chinese medicine preparation for the control of golden apple snail in rice that can be applied directly. Store in a sealed container away from light for later use.

5. The method for preparing the compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice according to claim 4, characterized in that, In S2, the fermentation temperature of the obtained traditional Chinese medicine fermentation liquid is 30℃~35℃, and the fermentation time is 7~10 days.

6. The method for preparing the compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice according to claim 4, characterized in that, In S2, the pH value of the fermentation broth for traditional Chinese medicine is 4.5~5.0, and the viable count is ≥2×10⁻⁶. 9 CFU / mL.

7. The method for preparing the compound traditional Chinese medicine preparation for controlling *Pomacea canaliculata* in rice according to claim 4, characterized in that, In S3, the conditions for performing vacuum negative pressure fine filtration are: temperature 30℃~35℃, vacuum degree 0.06MPa~0.07MPa, and feed rate 8~10L / h.

8. The method for preparing the compound traditional Chinese medicine preparation for controlling golden apple snails in rice according to claim 7, characterized in that, In S3, the transmittance of the filtrate after vacuum negative pressure filtration is ≥98%, and the retention rate of effective components is ≥95%.

9. The application of the compound traditional Chinese medicine preparation according to any one of claims 1-3 in the control of golden apple snails in rice, characterized in that, The compound traditional Chinese medicine preparation was applied to rice paddies.

10. The application according to claim 9, characterized in that, Includes the following steps: S100. One to two days before applying the pesticide, prepare the paddy field ditches and maintain a shallow water layer in the paddy field. The water depth should be controlled at 3cm to 5cm to ensure that the water surface of the paddy field evenly covers the soil surface, with no water accumulation dead corners or dry and cracked plots. S200. Dilute the prepared compound Chinese medicine preparation with water at a ratio of 1:150 to 1:200 and stir evenly. Quantitatively prepare the medicine according to the area of ​​the paddy field, using 30 kg to 40 kg of the medicine solution per mu of paddy field. S300. On a sunny day, between 9:00 and 11:00 in the morning or between 16:00 and 18:00 in the afternoon, use a backpack sprayer or drone to evenly spray the diluted compound Chinese medicine preparation on the entire paddy field. The water depth for spraying should be 3cm to 5cm. Focus on spraying the base of rice seedlings, the edge of the field ridges, and the areas around ditches where golden apple snails gather. S400: Seven days after the first application, a second application should be made to kill any remaining adult snails and newly hatched juvenile snails. A third preventative application should be made at the early tillering stage of rice. S500: Keep the shallow water layer in the paddy field without draining or changing the water for 3 days after application to ensure that the compound Chinese medicine preparation can fully exert its efficacy. Normal field water and fertilizer management can be carried out after 3 days.

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