COMPOUND PLANT-DERIVED MOLLUSCICIDE FOR CONTROLLING POMACEA CANALICULATA AND PREPARATION METHOD AND APPLICATION THEREOF

Abstract

A compound plant-derived molluscicide for controlling Pomacea canaliculata and a preparation method and application thereof are disclosed. The compound plant-derived molluscicide is prepared from following raw materials in proportion by weight: 40-45% of water extract of Solidago canadensis L. leaves, 45-50% of water extract of Lantana camara L. roots, 0.1-0.2% of active ingredient stabilizer, and 0.1-0.2% of preservative; and a balance is ultrapure water. The disclosure has rich raw material resources, low cost, good molluscicidal effect, sustained and stable efficacy, non-toxic effect on freshwater fish, and meets the requirements of environmental protection.

Claims

1. A compound plant-derived molluscicide prepared from following raw materials in proportion by weight: 40-45% of water extract of Solidago canadensis L. leaves; 45-50% of water extract of Lantana camara L. roots; 0.1-0.2% of active ingredient stabilizer; 0.1-0.2% of preservative; and the balance being ultrapure water.

2. The compound plant-derived molluscicide of claim 1, wherein the active ingredient stabilizer is Tween 80, and the preservative is sorbic acid or sodium benzoate.

3. The compound plant-derived molluscicide of claim 1, wherein a preparation method of the compound plant-derived molluscicide comprises: (1) collecting fresh Solidago canadensis L. leaves and fresh Lantana camara L. roots respectively, washing and draining; (2) cutting the above drained Solidago canadensis L. leaves and Lantana camara L. roots into 0.5-1 cm segments, respectively; (3) putting the Solidago canadensis L. leaf segments and the Lantana camara L. root segments prepared in step (2) into a 200-300 mesh crusher for crushing, respectively; (4) weighing the crushed Solidago canadensis L. leaf material and the crushed Lantana camara L. root material prepared in step (3) and adding with sterilized water in a volume ratio of 1:4-5 to each; putting the Solidago canadensis L. leaf material added with sterilized water and the Lantana camara L. root material added with sterilized water into a refrigerator at 4° C. overnight, grinding the Solidago canadensis L. leaf material and the Lantana camara L. root material after standing, and filtering the grinding liquid respectively with sterile gauze to obtain water extract of Solidago canadensis L. leaves and water extract of Lantana camara L. roots; (5) taking the water extract of Solidago canadensis L. leaves and the water extract of Lantana camara L. roots in a prescribed amount and mixing, adding sterilized water into the mixed water extract in a prescribed amount for dilution, and then adding with the active ingredient stabilizer and the preservative in a formula amount; after mixing evenly, standing overnight, and obtaining the compound plant-derived molluscicide by filtration.

4. A preparation method of the compound plant-derived molluscicide, comprising: (1) collecting fresh Solidago canadensis L. leaves and fresh Lantana camara L. roots respectively, washing and draining; (2) cutting the above drained Solidago canadensis L. leaves and Lantana camara L. roots into 0.5-1 cm segments, respectively; (3) putting the Solidago canadensis L. leaf segments and the Lantana camara L. root segments prepared in step (2) into a 200-300 mesh crusher for crushing, respectively; (4) weighing the crushed Solidago canadensis L. leaf material and the crushed Lantana camara L. root material prepared in step (3) and adding with sterilized water in a volume ratio of 1:4-5 to each; putting the Solidago canadensis L. leaf material added with sterilized water and the Lantana camara L. root material added with sterilized water into a refrigerator at 4° C. overnight, grinding the Solidago canadensis L. leaf material and the Lantana camara L. root material after standing, and filtering the grinding liquid respectively with sterile gauze to obtain water extract of Solidago canadensis L. leaves and water extract of Lantana camara L. roots; (5) taking the water extract of Solidago canadensis L. leaves and the water extract of Lantana camara L. roots in a formula amount and mixing, adding sterilized water into the mixed water extract in a formula amount for dilution, and then adding with the active ingredient stabilizer and the preservative in a formula amount; after mixing evenly, standing overnight, and obtaining the compound plant-derived molluscicide by filtration.

5. The preparation method of the compound plant-derived molluscicide of claim 1, wherein in the step (5), in terms of mass percentage, the formula amount of each component is: 40-45% of the water extract of Solidago canadensis L. leaves, 45-50% of the water extract of Lantana camara L. roots, 0.1-0.2% of the active ingredient stabilizer, and 0.1-0.2% of the preservative; and a balance is ultrapure water.

6. An application of the compound plant-derived molluscicide of claim 1 as a molluscicide for Pomacea canaliculate.

7. The application of claim 6, wherein the compound plant-derived molluscicide is formulated into a mixed agent with a concentration of 5-40 g/L and applied under 26-30° C.

Description

DETAILED DESCRIPTION OF EMBODIMENTS

[0023] In the following, the present disclosure will be further described with reference to specific embodiments, but the scope of the disclosure is not limited thereto.

Embodiment 1

[0024] Preparation of Water Extracts the Plants [0025] (1) Fresh Solidago canadensis L. leaves and fresh Lantana camara L. roots were collected respectively. After collection, they were washed and the surface water was drained. [0026] (2) The above drained Solidago canadensis L. leaves and Lantana camara L. roots were cut into 0.5-1 cm segments, respectively. [0027] (3) The Solidago canadensis L. leaf segments and the Lantana camara L. root segments prepared in step (2) were put into a 200-300 mesh crusher for crushing, respectively. [0028] (4) 50 g of each crushed plant material prepared in step (3) were weighed, and sterilized water in a volume ratio of 1:5 was added into each; each crushed plant material added with sterilized water was put into a refrigerator at 4° C. overnight; each plant material was ground after standing, and the grinding liquid was filtered with sterile gauze to obtain water extract of each plant.

Embodiment 2

[0029] Preparation of 1000 g Compound Plant-Derived Molluscicide

[0030] The formula was: 400 g of the water extract of Solidago canadensis L. leaves, 450 g of the water extract of Lantana camara L. roots, 1 g of the active ingredient stabilizer, 1 g of the preservative, and 148 g of ultrapure water.

[0031] The water extracts of Solidago canadensis L. leaves and Lantana camara L. roots were taken in the above formula amount, the formula amount of sterile water was added for dilution, then Tween 80 as the active ingredient stabilizer and the preservative (in this embodiment, it was sorbic acid) were added in the formula amount. The mixture was mixed evenly and stood overnight. Then the compound plant-derived molluscicide was obtained by filtration, and was put in a refrigerator for standby after labeling.

Embodiment 3

[0032] Preparation of 1000 g Compound Plant-Derived Molluscicide

[0033] The formula was: 450 g of the water extract of Solidago canadensis L. leaves, 500 g of the water extract of Lantana camara L. roots, 2 g of the active ingredient stabilizer, 2 g of the preservative, and 46 g of ultrapure water.

[0034] The water extract of Solidago canadensis L. leaves and Lantana camara L. roots were taken in the above formula amount, the formula amount of sterile water was added for dilution, then Tween 80 as the active ingredient stabilizer and the preservative (in this embodiment, it was sodium benzoate) were added in the formula amount. The mixture was mixed evenly and stood overnight. Then the compound plant-derived molluscicide was obtained by filtration, and was put in a refrigerator for standby after labeling.

Embodiment 4 Molluscicidal Test

[0035] 4.1 Molluscicidal Effect of the Two Water Extracts

[0036] 4.1.1 Pomacea canaliculata for Test

[0037] All the Pomacea canaliculata snails were collected from paddy fields and nearby ditches. The Pomacea canaliculata snails were collected, cleaned and domesticated in a 40 cm×25 cm×28 cm feeding vat in greenhouse. In the test, healthy Pomacea canaliculata snails with similar height and weight were selected as the test objects, and they were not fed during the test.

[0038] 4.1.2 Test solution: water extract of each plant prepared in embodiment 1. The water extracts of each plant were diluted into 40 g/L, 30 g/L, 20 g/L, 10 g/L and 5 g/L with ultrapure water. Meanwhile, ultrapure water was used as a blank control and niclosamide wettable powder was as a positive control.

[0039] 4.1.3 Test method: 600 ml of each component solution with concentration of 40 g/L, 30 g/L, 20 g/L, 10 g/L and 5 g/L was taken into a 1000 ml beaker, respectively. At the same time, ultrapure water was used as a blank control, and niclosamide wettable powder was used as a positive control. 20 Pomacea canaliculata snails were put into each beaker without feeding. The mouth of the beaker was covered with nylon net. The death number of Pomacea canaliculata snails in each beaker was checked every 24 hours, and the mortality rate of Pomacea canaliculata snails in different treatments at the corresponding time was calculated. The total observation time was 144 hours. The temperature was 26° C.±1° C. during the test. Three replicates were set for each treatment.

[0040] 4.1.4 Identification standard of Pomacea canaliculata: after putting Pomacea canaliculata snails into different concentrations of sample solution, the suspected dead snails in each treatment were taken out every 24 hours and put into dechlorinated water for observation. The snails floating on the water or suspending in the water within 24 hours without movement and response to external stimulation were regarded as dead snails.

[0041] 4.1.5 Data Processing and Analysis

[0042] Excel and SPSS 20.0 software were used to analyze the test data.

[0043] 4.1.6 Result Analysis

TABLE-US-00001 TABLE 1 Molluscicidal effect of water extracts of two plants Mortality rate (%) Poison time Group 24 h 48 h 72 h 96 h 120 h 144 h 40 g/L Water 36.7 ± 1.4a 52.3± 2.1a 55.7 ± 1.7a 61.3 ± 1.2b 82.3 ± 1.3b 91.0 ± 1.4b extract of Solidago canadensis L. leaves Water 41.3 ± 1.3a 50.3 ± 1.4a 61.7 ± 1.3b 76.4± 1.1b 85.0 ± 0b 85.0 ± 0 extract of Lantana camara L. roots 30 Water 28.3 ± 2.1a 31.7 ± 1.7a 53.3 ± 1.7a 61.7 ± 1.4b 80.3 ± 0b 94.3 ± 0b 30 g/L extract of Solidago canadensis L. leaves Water 38.3 ± 1.5a 45.3 ± 1.4a 56.7 ± 1.3a 70.4 ± 1.1b 84.3 ± 0b 85.0 ± 0b extract of Lantana camara L. roots 20 g/L Water 22.3 ± 2.2a 26.3 ± 1.6a 43.3 ± 1.3a 51.7 ± 1.8a 72.3 ± 1.2b 84.3 ± 1.2b extract of Solidago canadensis L. leaves Water 28.3 ± 1.7a 35.3 ± 1.3a 46.7 ± 1.4a 60.3 ± 1.2b 74.3 ± 1.5b 75.0 ± 0b extract of Lantana camara L. roots 10 g/L Water 15.3 ± 2.1c 16.3 ± 1.7c 23.3 ± 1.4a 51.3 ± 1.7a 62.3 ± 1.5b 74.3 ± 1.3b extract of Solidago canadensis L. leaves Water 19.3 ± 1.6c 30.3 ± 1.4a 45.7 ± 1.5a 50.3 ± 1.3a 64.3 ± 1.6b 65.0 ± 0b extract of Lantana camara L. roots  5 g/L Water  8.3 ± 1.1c 15.3 ± 1.5c 18.3 ± 1.4c 21.3 ± 1.7a 42.3 ± 1.6a 44.3 ± 1.4a extract of Solidago canadensis L. leaves Water 18.3 ± 1.6c 25.3 ± 1.4a 36.7 ± 1.5a 45.3 ± 1.3a 55.3 ± 1.6a 60.0 ± 0b extract of Lantana camara L. roots Bland control   0 ± 0a   0 ± 0 a   0 ± 0 a   0 ± 0 a   0 ± 0 a   0 ± 0 a Niclosamide wettable powder 64.8 ± 1.7b 80.3 ± 1.5b  100 ± 0c  100 ± 0c  100 ± 0c  100 ± 0c (1.25 mg/L)

[0044] It can be seen from Table 1 that the water extracts of two plants had certain molluscicidal effect on Pomacea canaliculata, and the mortality of Pomacea canaliculata increased with the increase of water extract concentration and treatment time. It can be seen from table 1 that the molluscicidal effect of the water extracts of two plants in a short time was not ideal. Although the mortality of Pomacea canaliculata in each group increased with the extension of treatment time, the molluscicidal effect was still weaker than that of the control drug niclosamide wettable powder (P<0.05). Within 24 hours, the water extract of Lantana camara L. roots with 40 g/L concentration had the best toxic effect on Pomacea canaliculata, and the mortality rate was 41.3%. Within 48 hours, the killing rate of 40 g/L water extracts of two plants to Pomacea canaliculata was more than 50%.

[0045] 4.2 Molluscicidal Effect of Compound Water Extract

[0046] The water extracts of two plants were mixed according to the formula amount (according to the formula amount of embodiment 2 and embodiment 3 respectively), and the molluscicidal effect of the water extracts of two plant was detected. The specific test method was the same as part 4.1 of embodiment 4. The results (Table 2) showed that the toxic effect of the mixture of water extracts of two plants on Pomacea canaliculata was higher than that of single water extract in corresponding concentration of (Table 1), and the mortality rate of Pomacea canaliculata increased with the extension of treatment time. Among all kinds of concentrations, 40 g/L corresponded to the best molluscicidal effect. The half mortality rate appeared within 24 hours, and the mortality rate of Pomacea canaliculata reached 100% within 72 hours. The molluscicidal effect was similar to that of niclosamide wettable powder.

TABLE-US-00002 TABLE 2 Molluscicidal effect of water extracts of two plants mortality rate (%) Poison time Group 24 h 48 h 72 h 96 h 120 h 144 h Embodiment 2 40 g/L 51.7 ± 1.4b 70.3 ± 2.1b  100 ± 0c  100 ± 0c  100 ± 0c  100 ± 0c 30 g/L 38.3 ± 2.1a 51.7 ± 1.7b 83.3 ± 1.7b 91.7 ± 1.4b  100 ± 0c  100 ± 0c 20 g/L 23.3 ± 1.1a 46.7 ± 1.4a 81.7 ± 1b 91.7 ± 1.6b  100 ± 0c  100 ± 0c 10 g/L 11.7 ± 1.7a 19.3 ± 1.2a 31.3 ± 1.3a 57.3 ± 1.8b 83.3 ± 1.7b  100 ± 0c  5 g/L  8.3 ± 1.7c 15.7 ± 1.4a 31.7 ± 1.5a   45 ± 0a 85.7 ± 1.1b 98.7 ± 1.7c Embodiment 3 40 g/L 55.3 ± 1.7b   75 ± 1.4b  100 ± 0c  100 ± 0c  100 ± 0c  100 ± 0c 30 g/L 45.7 ± 1.3b 58.7 ± 1.8b 88.3 ± 1.2b  100 ± 0c  100 ± 0c  100 ± 0c 20 g/L 37.7 ± 1.7a 48.3 ± 1.4b 83.3 ± 1.5b 96.3 ± 1.4c  100 ± 0c  100 ± 0c 10 g/L 16.7 ± 1.6a 40.0 ± 0b 38.3 ± 1.5b 63.3 ± 1.7b 86.7 ± 1.7b  100 ± 0c  5 g/L 15.3 ± 1.8a 22.3 ± 1.3a 34.3 ± 1.2a 45.0 ± 0b 81.7 ± 1.5b  100 ± 0c Blank control   0 ± 0a   0 ± 0 a   0 ± 0 a   0 ± 0 a   0 ± 0 a   0 ± 0 a (ultrapure water) Niclosamide wettable 64.7 ± 2.4b 80.3 ± 1.7b  100 ± 0c  100 ± 0c  100 ± 0c  100 ± 0c powder (1.25 mg/L)

Embodiment 5

[0047] 5.1 Test Animals

[0048] All the Pomacea canaliculata snails were collected from paddy fields and nearby ditches. Pomacea canaliculata snails was collected, cleaned and domesticated in a 40 cm×25 cm×28 cm feeding vat in greenhouse. In the test, healthy Pomacea canaliculata snails with similar height and weight were selected as the test objects, and they were not fed during the test.

[0049] 5.2 Test Solution

[0050] The compound plant-derived molluscicide with a concentration of 40g/L, which was prepared in embodiment 2 and embodiment 3 and proved to be highly toxic to Pomacea canaliculata by test, was as the test solution. Meanwhile, ultrapure water was used as blank control.

[0051] 5.3 Test method: 600 ml of the test solution with concentration of 40 g/L was taken into a 1000 ml beaker. At the same time, ultrapure water was used as a blank control. 20 Pomacea canaliculata snails were put into each beaker without feeding. The mouth of the beaker was covered with nylon net. The death number of Pomacea canaliculata snails in each beaker was checked every 24 hours, and the mortality rate of Pomacea canaliculata snails in different treatments at the corresponding time was calculated. The total observation time was 144 hours. The temperature was 26° C.±1° C. during the test. Three replicates were set for each treatment.

[0052] At the same time, 600 ml of the test solution with concentration of 40 g/L was taken into a 1000 ml beaker, and ultrapure water was used as a blank control. 20 healthy red carp (7-8cm in length) were put into each beaker and raised as usual. The death number of red carp in each beaker was checked every 24 hours. The total observation time was 72 hours. The temperature was 26° C.±1° C. during the test. Three replicates were set for each treatment.

TABLE-US-00003 TABLE 3 Toxicity test of freshwater fish mortality rate % Poison time Group 24 h 48 h 72 h Red carp 0 0 0 Embodiment 2 51.7 ± 1.4 70.3 ± 2.1 100 ± 0 (40 g/L) Embodiment 2 55.3 ± 1.7   75 ± 1.4 100 ± 0 (40 g/L) Blank control (ultrapure water)   0 ± 0a    0 ± 0a    0 ± 0a