BAM15-containing drug for killing or preventing and treating ampullaria gigas and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly discloses a medicament for preventing or killing ampullaria gigas and application of BAM15 in prevention or killing of ampullaria gigas. The application of the BAM15 in killing or preventing and treating the ampullaria gigas is evaluated, and by observing the killing effects of BAM15 with different doses on the ampullaria gigas, the BAM15 is proved to have an obvious effect of killing the ampullaria gigas, so that the BAM15 can be widely applied to the field of prevention and treatment of the ampullaria gigas.

Description

TECHNICAL FIELD

[0001] The application belongs to a new use of BAM15, and particularly relates to an application, a medicament and a method for preventing or killing Pomacea canaliculata by using BAM15 as a molluscicide. BACKGROUND

[0002] Pomacea canaliculata, also known as apple snail, belongs to Gastropoda, Mesogastropoda and Ampullariidae, and is a freshwater gastropod originally from South America. It is widely distributed in the world, especially in tropical and subtropical regions. Due to its strong reproductive capacity and adaptability, it has become a global invasive species. In China, there are two species of Pomacea canaliculata and Pomacea maculata. Due to its large size, varied diet, fast growth, strong reproductive capacity and wide adaptability, Pomacea canaliculata not only causes great losses to crop planting such as rice in China, but also destroys the ecological environment and seriously threatens human health, and has become an important object of prevention and control in agricultural production.

[0003] As an important aquatic pest, Pomacea canaliculata is commonly found in rice fields, lakes and rivers. In the ecological system, the presence of Pomacea canaliculata leads to the reduction of aquatic plants, thereby affecting the ecological balance of the water body. In agricultural production, Pomacea canaliculata causes direct harm to crops such as rice, affecting the economic income of farmers. In addition, Pomacea canaliculata is also an intermediate host of many parasites, especially Echinostoma revolutum and Angiostrongylus cantonensis. These parasites can infect humans and other animals through Pomacea canaliculata, causing serious public health problems. Therefore, Pomacea canaliculata has caused serious threats to the ecological system, agricultural production and public health safety.

[0004] At present, there are three main methods for preventing and killing Pomacea canaliculata, namely physical prevention, biological prevention and chemical prevention. Although physical prevention and biological prevention are harmless to the environment, they have low molluscicidal efficiency, limited popularization, time-consuming and labor-intensive, and are easy to cause crop reduction. At present, the prevention of Pomacea canaliculata mainly relies on chemical prevention. Although chemical prevention has significant effect and saves time and labor, it has the problems of high toxicity to non-target organisms and the possibility of increasing the resistance of Pomacea canaliculata to drugs due to long-term use. Traditional chemical molluscicides mainly include niclosamide, metaldehyde and fentin acetate. Niclosamide is the only molluscicide recommended by the World Health Organization. Studies have found that 70% niclosamide wettable powder is applied in the field (35 g / 667 m 2) control the golden apple snail, the control effect is as high as 96%, but it also has a strong toxic effect on fish. Tetramethylaldehyde can not only control harmful creatures such as land snails and slugs, but also has a good control effect on golden apple snails. Zhan Zhixiong et al. reported that 6% tetramethylaldehyde granules with soil scattering (500-700 g / 667 m2) have a control effect on golden apple snails of more than 80%. Tin phenylacetate, also known as baipaiwei. Li Renke et al. found that 45% baipaiwei super-micro wettable powder with soil scattering in the field (35 g / 667 m 2 ) has a control effect on golden apple snails of 92.7% in 3 days. In addition, some new compounds have been found to have the potential to become snail killers, such as PBQ ([1-(4.chlorophenyl)-3-(pyridin-3- yl)urea]), and the salt of quinoid-2',5-dichloro-4'-nitro-salicylanilide. Although chemical control has good control effect, there are few chemical snail killers, which can easily make snails resistant, and can also kill fish and other non-target organisms, and long-term use can destroy water biodiversity, cause environmental pollution and agricultural product safety problems. After some snail killers are applied in water, the golden apple snails also escape from the water surface in time, thereby reducing the snail killing effect.

[0005] Therefore, developing a new type of snail killer with high efficiency, environmental compatibility and sensitivity has broad application prospects.

[0006] BAM15, also known as CS-2890, is an organic compound with the chemical name N5,N6-bis(2-fluorophenyl)-[1,2,5]oxadiazolo[3,4-B]pyrazine-5,6-diamine, which is a white to beige powder, and the molecular formula is C 16 H 10 F2N6O, which is dissolved in dimethyl sulfoxide (DMSO). BAM15, as a new type of mitochondrial proton uncoupler, has the potential to regulate mitochondrial function and energy metabolism, and shows good application prospects in the treatment of diseases such as obesity, acute kidney injury and tumors. However, there is no public report on the application of BAM15 to kill golden apple snails. On the basis of extensive drug screening, the applicant accidentally found that BAM15 has drug activity of killing golden apple snails. In view of this, the present application provides the application, medicament and method of BAM15 in preventing or killing golden apple snails. SUMMARY

[0007] The technical problem to be solved by the present application is to provide the application, medicament and method of BAM15 in preventing or killing golden apple snails.

[0008] The technical solution of the present application to solve the above technical problems is as follows: A medicament for killing or controlling the golden apple snail, the medicament containing BAM15.

[0009] Further, the dosage form of the medicament is selected from any one of the following: suspension, emulsion, microemulsion, wettable powder.

[0010] Further, the medicament further comprises a pesticide-acceptable adjuvant.

[0011] Further, the adjuvant comprises a penetrating agent DMSO.

[0012] In addition, the present application also provides the use of BAM15 in controlling or killing the golden apple snail.

[0013] Further, the present application uses BAM15 to control or kill the golden apple snail.

[0014] The present application also provides a method for controlling or killing the golden apple snail, comprising the following steps: applying the above-mentioned medicament for controlling or killing the golden apple snail into the water body.

[0015] Further, the application amount of the medicament is such that the concentration of the effective component of the medicament in the water body is greater than 0.1142 mg / L.

[0016] Further, the immersion killing time of the medicament is 24-72 hours.

[0017] The beneficial effects of the present application are: the present application evaluates the application of BAM15 in controlling or killing the golden apple snail, and through observation of the killing effect of different doses of BAM15 on the golden apple snail, it is confirmed that BAM15 has obvious snail-killing effect, and therefore it is possible to be widely applied in the field of golden apple snail control. DETAILED DESCRIPTION

[0018] The present application takes BAM15 as the research object, and explores the effect of BAM15 in killing the golden apple snail, and discovers a new application field of BAM15.

[0019] Snail-killing experiment example 1: effect of BAM15 in killing the golden apple snail BAM15 as experimental material, DMSO solution to help BAM15 dissolved in water, according to the experimental needs of 1 mg / L, 0.5 mg / L, 0.25 mg / L, 0.125 mg / L, 0.0625 mg / L of the drug solution, set the dechlorinated water as a blank control group. Each dose set three repeats, take the immersion method, 400 mL of liquor poured into a 500 mL beaker. Select different size of healthy active apple snails, specifically 2-3 mm, 10-15 mm and 20-25 mm of three groups, each beaker put in 5 snails, covered with breathable gauze, tied with rubber band, to prevent the snails escape. The specific experimental process is as follows.

[0020] 1. Experimental reagents and materials 1.1 Experimental reagents BAM159 with a purity of 99% (commercially available from Shanghai Maikelin Biochemical Technology Co., Ltd.), DMSO.

[0021] 1.2 Materials Apple snails: To reduce experimental errors, apple snail populations were established in the laboratory in advance. In 2021, 20 apple snail eggs were collected from Wuxi Shangxianhe Wetland Park (31.525981 °N, 120.304034 °E) in Jiangsu Province. The eggs were incubated in a plastic box (40 cm long x 45 cm wide x 40 cm high) containing dechlorinated water at room temperature. The hatched snails were raised at room temperature and fed with lettuce leaves under natural light. All apple snails were fed once a day. To maintain water quality, the water in the tank was changed at least every 7 days. Healthy and active apple snails of three different sizes, 2-3 mm, 10-15 mm and 20-25 mm, were selected.

[0022] 1.3 Experimental instruments and equipment Several 500 mL beakers, breathable gauze, rubber band, pipette, electronic balance, weighing paper, thermometer, forceps, ep tube, needle.

[0023] 2. Experimental method 2.1 Killing effect of BAM15 on apple snails Liquid drug was prepared with 99% pure BAM15, DMSO and dechlorinated water. The concentrations of the experimental group were 1 mg / L, 0.5 mg / L, 0.25 mg / L, 0.125 mg / L and 0.0625 mg / L. The blank control was dechlorinated water and DMSO. Three different sizes of 2-3 mm, 10-15 mm and 20-25 mm of golden apple snails were transferred into 500 mL beakers containing 400 mL of test solution with different concentrations (1 mg / L, 0.5 mg / L, 0.25 mg / L, 0.125 mg / L, 0.0625 mg / L), 5 golden apple snails were placed in each beaker, and the beaker was sealed with gauze to prevent the golden apple snails from escaping. Each concentration was set up for 24 h, 48 h and 72 h groups, and the golden apple snails were taken out after immersion and then the mortality rate was evaluated. The golden apple snails were declared dead when the snail body was completely retracted, the antennae had no response, and the needle had no movement response. The mortality rate was used to calculate the LC 50 value. The mortality rate was recorded and calculated using the following formula:

[0024] 3. Data processing and analysis All statistical analysis was performed using Graphpad Pism 9.5 software. Nonlinear curve fitting was performed on the mortality data to calculate the LC 50 concentration.

[0025] 4. Experimental results Example 1: Effect of BAM15 on golden apple snails The mortality rate of golden apple snails in the experimental treatment increased with the increase of treatment time or BAM15 concentration. Although the LC 50 of 10-15 mm golden apple snails for 24 h could not be calculated, the mortality rate was significantly positively correlated with the concentration, indicating that the drug had a dose-dependent effect. The LC 50 increased with the increase of snail size (e.g., the LC 50 of 2-3 mm golden apple snails was significantly lower than that of 20-25 mm golden apple snails), indicating that BAM15 was more sensitive to 2-3 mm golden apple snails and had a higher efficient target killing effect (Table 1).

[0026] Table 1 Effect of different concentrations of BAM15 on golden apple snails at different times

[0027] Example 2 Preparation of emulsion Oil phase preparation: 20g of ethyl acetate and 25g of acetone were added to a 500mL three-necked flask, stirring was started (rotation speed 300-500r / min), after mixing, 20g of BAM15 was slowly added, stirring was continued (rotation speed 500-800r / min), the temperature was controlled at 25-30 DEG C, and stirring was continued for 30-40 minutes until the BAM15 was completely dissolved, to obtain a clear oil agent, for standby.

[0028] Water phase preparation: 30g of deionized water was added to a 300mL beaker, heated to 40-50 DEG C in a constant temperature water bath, 5g of benzyl phenol polyoxyethylene polyoxypropylene ether, 3g of phenethyl phenol polyoxyethylene ether formaldehyde condensate (mass ratio of the two emulsifiers 5:3), 4g of polyvinyl alcohol (polymerization degree 1750±50), 5g of propylene glycol, 0.5g of silicone defoaming agent were added in sequence, stirring was started (rotation speed 600-800r / min), and the mixture was stirred for 40-50 minutes until all the materials were completely dissolved, to form a uniform transparent auxiliary composite water agent, which was cooled to room temperature for standby.

[0029] Emulsification process: the auxiliary composite water agent was transferred to a 1000mL emulsification tank equipped with a high shear emulsifier, the emulsifier was started, and pre-stirring was carried out at 800r / min for 1-2 minutes, then under high speed shearing (rotation speed controlled at 1000r / min), the oil agent was slowly added into the water phase through a constant pressure dropping funnel (dropping speed 2-3mL / min), after the addition was completed, the shearing was continued at 1000r / min for 60 minutes, and the temperature of the system was controlled at 25-30 DEG C, finally a milky white opaque liquid was obtained, which was a drug emulsion.

[0030] Quality detection: a small amount of emulsion was taken, and the appearance was uniform milky white without stratification; the average particle size of the emulsion droplets was 1-5mu measured by a laser particle size analyzer; 50mL of the emulsion was taken in a centrifuge tube, and centrifuged at 3000r / min for 30 minutes, without stratification and demulsification, to obtain a qualified product.

[0031] As can be seen from the above, the application of BAM15 for the prevention and killing of golden apple snails is evaluated, by observing the killing effect of different doses of BAM15 on golden apple snails, it is confirmed that BAM15 has obvious snail killing effect, and therefore can be widely applied in the field of golden apple snail prevention.

Claims

1. An agent for killing or controlling the golden apple snail, characterized by The agent contains BAM15.

2. The agent for preventing or killing the golden apple snail according to claim 1, wherein The dosage form of the agent is selected from any one of the following: suspension, emulsion, microemulsion, wettable powder.

3. The agent for preventing or killing the golden apple snail according to claim 2, wherein The agent further comprises a pesticide-acceptable adjuvant.

4. The agent for preventing or killing the golden apple snail according to claim 3, wherein The adjuvant comprises a penetrating agent DMSO.

5. Use of BAM15 in preventing or killing the golden apple snail.

6. Use of BAM 15 according to claim 5 for the control or killing of P. canaliculata, characterized in that, BAM15 is used in preventing or killing the golden apple snail.

7. A method of controlling or killing a golden apple snail, characterized by, The method comprises the following step: applying an agent for preventing or killing the golden apple snail according to any one of claims 1-4 into a water body.

8. A method of controlling or killing a golden apple snail according to claim 7, characterized in that, The agent is applied in an amount such that the concentration of the active ingredient of the agent in the water body is greater than 0.1142 mg / L.

9. A method of controlling or killing the golden apple snail according to claim 7, characterized in that, The immersion killing time of the agent is 24-72 hours.

Images