Application of pulegone in pest control

Abstract

The present invention provides an application of pulegone in pest control, and relates to the technical field of pest control. The pulegone can effectively kill adult golden apple snails at a high concentration, and can inhibit the laying of eggs of golden apple snails and reduce the hatching rate of eggs laid by golden apple snails at a low concentration. At the same time, spraying golden apple snail egg masses at a low concentration can effectively inhibit the hatching of eggs and reduce the hatching rate. That is, the use of pulegone, a plant-derived component, can kill golden apple snails, and can also effectively destroy the reproductive reproduction of golden apple snails, further realizing the prevention and control of golden apple snails, and on this basis, pulegone can also kill harmful snails such as striped snails.

Description

Technical Field

[0001] The present invention relates to the technical field of pest control, and in particular to the application of pulegone in pest control. Background Art

[0002] The prevention and control of golden apple snails should adopt the strategy of "prevention first, comprehensive prevention and control". Among them, chemical prevention and control is still the main measure for the prevention and control of golden apple snails due to its advantages of fast effect and good effect. Currently, the widely used snailicides include 6% tetraacetaldehyde granules, 70% snailicide ethanolamine salt wettable powder, 6% polyaldehyde·carbaryl granules, 70% snailicide amine wettable powder and other chemical products. Although chemical snailicides have a significant control effect on golden apple snails in a short period of time, there are fewer varieties of chemical snailicides, which can easily make golden apple snails resistant to drugs and poison fish and other non-target organisms. Long-term use will not only destroy the biodiversity of aquatic areas, but also cause environmental pollution and agricultural product safety problems. At the same time, after some chemical snailicides are applied in water bodies, golden apple snails will escape from the water in time, thereby reducing their snail-killing effect.

[0003] As a compound with a specific chemical structure, pulegone has certain insecticidal activity against insects. In existing studies, it is generally used to kill insects such as the rusty red flour beetle, and the overall killing effect is poor. It is usually used as an auxiliary additive to work together with other insecticidal ingredients. There is no research or report on the killing and prevention of mollusks such as snails and snails by pulegone in current research and public documents. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention provides an application of pulegone in pest control, that is, pulegone, a plant-derived ingredient, is used to kill the golden apple snails, and at the same time, it can also effectively destroy the reproductive reproduction of the golden apple snails, further achieving the control of the golden apple snails, and on this basis, pulegone can also kill harmful snails such as the striped Chinese snail.

[0005] To achieve the above objectives, the technical solution of the present invention is implemented through the following technical solutions:

[0006] Pulegone is used to kill harmful organisms or destroy their reproduction.

[0007] Preferably, the harmful organisms are molluscs.

[0008] Preferably, the harmful organism is Pomacea canaliculata or Pomacea fasciatus.

[0009] Preferably, the LC values ​​of pulegone on Pomacea canaliculata at 24 h, 48 h, 72 h, 96 h, 120 h and 144 h are 50They are 542.00 mg / L, 182.23 mg / L, 55.88 mg / L, 49.45 mg / L, 45.83 mg / L and 43.38 mg / L respectively.

[0010] Preferably, the LC values ​​of pulegone on snails at 24 h, 48 h, 72 h and 96 h are 50 They are 8.74 mg / mL, 7.23 mg / mL, 6.59 mg / mL and 6.45 mg / mL respectively.

[0011] Preferably, the specific manner in which the pulegone is used to destroy the reproduction and propagation of harmful organisms is to inhibit the spawning of Pomacea canaliculata snails or inhibit the hatching of Pomacea canaliculata snail eggs.

[0012] Preferably, the concentration of pulegone for inhibiting the spawning of Pomacea canaliculata is 20-40 mg / L, and the treatment time is more than 2 h.

[0013] Preferably, the concentration of pulegone for inhibiting the hatching of Pomacea canaliculata eggs is 30-60 mg / L, and the treatment method is direct spraying.

[0014] A product for inhibiting the hatching of Pomacea canaliculata eggs, wherein the concentration of pulegone in the product is above 30 mg / L.

[0015] The present invention provides the use of pulegone in pest control, which has the following advantages compared with the prior art:

[0016] The present invention adopts pulegone to kill the golden apple snails, and the golden apple snails can be killed at a higher concentration, and the reproduction of the golden apple snails can be destroyed at a lower concentration, so that the egg-laying rate of the golden apple snails can be significantly reduced, and the hatching rate of the eggs after the golden apple snails lay eggs can also be reduced. At the same time, directly treating the golden apple snail eggs with a low concentration can also further reduce the hatching rate of the eggs, thereby providing an effective means for the biological control of the golden apple snails. In addition, pulegone can also kill the striped snails, thereby achieving the purpose of preventing and controlling multiple harmful organisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a schematic diagram of an adult Pomacea canaliculata in Example 1 of the present invention;

[0018] Figure 2 This is a schematic diagram of laboratory culture of Pomacea canaliculata in Example 1 of the present invention;

[0019] Figure 3 This is a schematic diagram of the Pomacea canaliculata snails being placed in a plastic box and killed by immersion in Example 1 of the present invention;

[0020] Figure 4 This is a schematic diagram of washing the Pomacea canaliculata snails killed by immersion in Example 1 of the present invention with dechlorinated water;

[0021] Figure 5 This is a schematic diagram of a surviving golden apple snail in Example 1 of the present invention;

[0022] Figure 6 This is a schematic diagram of a dead golden apple snail in Example 1 of the present invention;

[0023] Figure 7 This is a schematic diagram of the spawning of the golden apple snail in Example 1 of the present invention. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention is clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Embodiment 1:

[0025] 1. Verify the killing effect of pulegone on Pomacea canaliculata

[0026] 1. Raw material acquisition:

[0027] Pulegone (purchased from Shanghai Yuanye Biotechnology Co., Ltd.) was used as the experimental compound, metaldehyde (purchased from Henan Jubang Biopharmaceutical Co., Ltd.) was used as the control compound; adult Pomacea canaliculata (purchased from Rong County, Zigong City, Sichuan Province) was used as the experimental material (such as Figure 1 shown);

[0028] 2. Experimental design:

[0029] (1) Treatment of test organisms:

[0030] The snails were cultured in plastic basins (60 cm in diameter and 30 cm in height) filled with tap water (see Figure 2 As shown), covered with nylon nets, and placed in the laboratory for breeding. Maintain natural light, change water regularly every day, and clean up dead snails. Raise them under laboratory conditions to adapt them to the laboratory environment. Before conducting the experiment, use the water culture method to test the adaptability of the golden apple snails to the laboratory environment (put the golden apple snails in dechlorinated water, the golden apple snails that do not float can be used for subsequent experiments, and the golden apple snails that float are dead), and select golden apple snails of the same size for the experiment;

[0031] (2) Preparation of test solution:

[0032] According to the characteristics of the compound, weigh or measure the corresponding amount of the compound and place it in a beaker, add 750 ml of 0.5% Tween 80 aqueous solution to prepare pulegone and metaldehyde (positive control) solutions.

[0033] (3) Determination of snail-killing activity:

[0034] The molluscicidal activity of pulegone against Pomacea canaliculata was determined by the immersion method. Five concentration gradients of pulegone solutions of 20 mg / L, 40 mg / L, 80 mg / L, 160 mg / L and 320 mg / L and five concentration gradients of metaldehyde solutions of 10 mg / L, 20 mg / L, 40 mg / L, 80 mg / L and 160 mg / L were prepared, each concentration was repeated 3 times, and the molluscicidal activity was determined.

[0035] Number the plastic boxes (21 cm long × 15 cm wide × 7 cm high) and mark the concentrations, then pour the corresponding test solutions into them. Add 750 mL of the test solution into each plastic box, put 10 selected apple snails in, and cover them with lids to prevent the snails from crawling out (e.g. Figure 3 After immersion for 24 h, 48 h, 72 h, 96 h, 120 h, and 144 h, pour out the solution and rinse with dechlorinated water three times (as shown in Figure 4 The snails were placed in a new plastic box for 72 hours and observed every 24 hours. Figure 5 As shown in the figure, the dead golden apple snails are Figure 6 As shown in the table above), record the number of dead snails and the total number of remaining snails in each treatment each day. Death criteria for Pomacea canaliculata: Place Pomacea canaliculata snails in dechlorinated water. If they float, they are dead. If they have their opercula tightly closed and the snail flesh collapses when lightly touched with tweezers, they are dead.

[0036] (4) Data processing:

[0037] The adjusted mortality rate was calculated using the Abbott correction formula, with the mortality rate in the control group being <20%.

[0038]

[0039]

[0040] SPSS 23.0 statistical analysis software was used for univariate Anova test analysis to obtain the mean ± standard error of pulegone and the positive control metaldehyde; regression probability test analysis was performed to obtain the median lethal concentration LC50, 95% confidence interval of LC50 value, slope ± standard error, chi-square value ( X 2 ) and P value.

[0041] The corrected mortality of pulegone and the positive control metaldehyde on Pomacea canaliculata is shown in Table 1:

[0042] Table 1

[0043]

[0044] As can be seen from Table 1, pulegone and the positive control metaldehyde have different degrees of snail killing activity against Pomacea canaliculata. The corrected mortality of pulegone at the concentration of 160 and 320 mg / L on the second day was greater than or equal to 50%; the corrected mortality of pulegone at the concentration of 40, 80, 160, and 320 mg / L on the third, fourth, fifth, and sixth days was greater than or equal to 50%. The corrected mortality of the positive control metaldehyde at the concentration of 160 mg / L on the second day was greater than 50%; the corrected mortality of 40, 80, and 160 mg / L on the third, fourth, fifth, and sixth days was greater than 50%; among them, the corrected mortality of 40, 80, and 160 mg / L on the sixth day reached 100%.

[0045] The snail-killing activity of pulegone and the positive control metaldehyde against Pomacea canaliculata is shown in Table 2. Based on the data in Table 1, regression probability test analysis was performed using SPSS software to calculate the median lethal concentration (LC) of pulegone and the positive control metaldehyde against Pomacea canaliculata at 1, 2, 3, 4, 5 and 6 days, respectively. 50 , LC 50 95% confidence interval, slope ± standard error, chi-square value ( X 2 ) and P value.

[0046] Table 2

[0047]

[0048] It can be seen from Table 2 that on the 1st and 2nd day, the snail-killing activity of pulegone against Pomacea canaliculata was not as good as that of the positive control metaldehyde. The LC 50 The values ​​were 238.81 and 110.93 mg / L respectively; the LC 50 The values ​​were 542.00 and 183.23 mg / L, respectively. On the third day, the snail-killing activity of pulegone against Pomacea canaliculata was also not as good as that of the positive control metaldehyde. The LC 50 The value was 42.10 mg / L; however, the activity of pulegone sarcosyltransferase was increased, and the LC 50 The value was 55.88 mg / L; at 4, 5, and 6 days, the LC of pulegone 50 The values ​​were 49.45, 45.83, and 43.38 mg / L respectively.

[0049] 2. Verify the damage of pulegone to the reproduction and reproduction of golden apple snails:

[0050] 1. Verify the effect of pulegone on the spawning rate of Pomacea canaliculata:

[0051] (1) Preparation of test solution:

[0052] Pulegone solutions with concentrations of 0 mg / L, 15 mg / L, 20 mg / L, 30 mg / L, 40 mg / L, and 45 mg / L were prepared.

[0053] (2) Treatment of test organisms:

[0054] The snails were cultured in a plastic basin (60 cm in diameter and 30 cm in height) filled with a certain volume of tap water, covered with a nylon net, and placed in the laboratory for breeding. Natural light was maintained, and the water was changed regularly every day, and dead snails were cleaned up. The snails were raised under laboratory conditions to adapt to the laboratory environment, and then snails of the same size were selected for the experiment;

[0055] (3) Determination of egg-laying rate of Pomacea canaliculata:

[0056] Single factor experiment (experiment start date is May 10, 2024):

[0057] S1. Determine the effect of different concentrations of pulegone on the spawning of Pomacea canaliculata snails:

[0058] Pour pulegone solutions of different concentrations into plastic boxes (length 21 cm × width 15 cm × height 7 cm), 750 mL per box, 10 boxes for each concentration, 5 pairs of adult male and female Pomacea canaliculata snails were placed in each box, soaked for 4 hours, then taken out and placed in plastic buckets (diameter 25 cm × height 60 cm), observe and collect mated female snails, select 20 from each group, place them in plastic buckets and continue to feed them with clean water for 15 days, set a net cover on the top of the plastic bucket to allow the Pomacea canaliculata snails to lay eggs on the inner wall of the plastic bucket (egg laying pictures as shown in the figure). Figure 7 The egg-laying rate of female snails in each barrel at 3d, 7d, and 15d was observed and recorded (egg-laying rate = number of egg-laying female snails / total number of female snails × 100%). The specific results are shown in Table 3 below:

[0059] Table 3

[0060]

[0061] As can be seen from the above table, the spawning rate of Pomacea canaliculata began to decline after treatment with 15 mg / L pulegone solution, but the spawning rate was as high as 90% after 15 days, and the overall decline was not large. When the concentration of pulegone solution was 20 mg / L and above, the spawning rate of Pomacea canaliculata dropped significantly, and the 15-day spawning rate was less than 60%. When the concentration of pulegone solution was 45 mg / L, its spawning rate was basically unchanged compared with 40 mg / L. Based on the principle of economy, the concentration of 20 mg / L-40 mg / L is the optimal treatment concentration.

[0062] Detection of the effect of the treatment on the hatching rate of Pomacea canaliculata eggs:

[0063] The eggs laid by the Pomacea canaliculata treated with the above different concentrations were collected and incubated at 27±2°C. The hatching rate after 15 days was recorded (hatching rate = number of Pomacea canaliculata eggs hatched / total number of Pomacea canaliculata eggs × 100%). The specific results are shown in Table 4 below:

[0064] Table 4

[0065]

[0066] As shown in Table 4 above, the hatching rate of eggs laid by Pomacea canaliculata treated with 20, 30, 40, and 45 mg / L pulegone solutions was reduced to below 65%.

[0067] S2. Determine the effect of different treatment times at the same concentration on the spawning of Pomacea canaliculata:

[0068] Pulegone solution with a concentration of 30 mg / L was poured into plastic boxes (length 21 cm × width 15 cm × height 7 cm), 750 mL per box, and a total of 60 boxes were set up. Five pairs of adult male and female Pomacea canaliculata snails were placed in each box and divided into 6 groups on average. They were soaked for 1 h, 1.5 h, 2 h, 3 h, 4 h and 5 h in sequence, and then taken out and placed in plastic buckets (diameter 25 cm × height 60 cm). The mated female snails were observed and collected. 20 snails were selected from each group and placed in plastic buckets with clean water for 15 days. The egg-laying rates of female snails in each bucket at 3d, 7d and 15d were observed and recorded. The specific results are shown in Table 5 below:

[0069] Table 5

[0070]

[0071] It can be seen from Table 5 above that when the immersion time is 1h and 1.5h, the overall spawning rate decreases slightly and is still above 80%. When the immersion time is 2h, 3h, 4h, and 5h, the spawning rate finally drops to below 60%, and the spawning rate also decreases as the immersion time increases.

[0072] Detection of the effect of the treatment on the hatching rate of Pomacea canaliculata eggs:

[0073] The eggs laid by the Pomacea canaliculata treated with different immersion times were collected and incubated at 27±2°C. The hatching rate after 15 days was recorded. The specific results are shown in Table 6 below:

[0074] Table 6

[0075]

[0076] It can be seen from Table 6 above that with the increase of immersion time, the hatching rate after spawning gradually decreases, and the hatching rate is lower than 65% at a time of 2h or more.

[0077] 2. Verify the effect of pulegone on the hatching rate of Pomacea canaliculata eggs

[0078] (1) Preparation of test solution:

[0079] Pulegone solutions with concentrations of 0 mg / L, 25 mg / L, 30 mg / L, 40 mg / L, 50 mg / L, 60 mg / L, and 65 mg / L were prepared.

[0080] (2) Treatment of test organisms:

[0081] The snails were cultured in a bucket (30 cm in diameter and 60 cm in height) filled with tap water, covered with a nylon net, and placed in a laboratory for breeding. Natural light was maintained, and the water was changed regularly every day, and dead snails were cleaned up. The snails were raised under laboratory conditions, the spawning situation was observed, and the normal snail egg masses were promptly removed for use;

[0082] (3) Egg mass hatching rate detection:

[0083] 21 egg masses of Pomacea canaliculata were taken and divided into 7 groups, with 3 egg masses in each group. Different concentrations of pulegone solution were sprayed, and the spraying amount was 1 / 10 of the total mass of the egg mass. The eggs were then incubated at 27±2°C, and the hatching rate was recorded after 15 days. The specific results are shown in Table 7 below:

[0084] Table 7

[0085]

[0086] It can be seen from Table 7 above that treating the egg masses of Pomacea canaliculata with pulegone at concentrations of 30, 40, 50, 60, and 65 mg / L can effectively reduce the hatching rate to less than 50%, and the hatching rate at 65 mg / L is less changed than that at 60 mg / L. Based on cost control considerations, it is more economical to treat Pomacea canaliculata eggs with pulegone at a concentration of 30-60 mg / L, and the hatching rate of the eggs is lower. Embodiment 2:

[0087] Verify the killing effect of pulegone on snails:

[0088] 1. Test organisms: Cathaica fasciola, purchased online in March 2024, produced in Xichuan County, Henan Province, fed in the laboratory before the experiment, regularly fed with lettuce, and regularly replaced the leaves and boxes;

[0089] 2. Preparation of test solution:

[0090] According to the characteristics of the compound, weigh or measure the corresponding amount of the compound and place it in a beaker, add 750 ml of 0.5% Tween 80 aqueous solution to prepare pulegone and metaldehyde (positive control) solution;

[0091] 3. Toxicity test on snails:

[0092] The stomach poisoning experiment on snails was conducted by leaf immersion method. Pulegone and positive control metaldehyde were set to 1.25, 2.5, 5, 10 and 20 mg / mL gradient concentrations. Ten snails were inoculated with each concentration. Each treatment was repeated 3 times, and a blank control was set. Different concentrations of the drug solution were poured into a container with a capacity of 10 mL, and the lettuce was soaked for 10 s before being fed to the snails. The death of the snails was observed at 24, 48, 72 and 96 h.

[0093] 4. Data processing:

[0094] SPSS 23.0 software was used to perform one-way analysis of variance (Anova), and the adjusted mortality rate, LC50 value and 95% confidence interval were calculated according to formulas (2-1) and (2-2). The chi-square test (χ2) was used to evaluate the goodness of fit of the virulence regression model.

[0095]

[0096] The corrected mortality rates of the snails treated with different concentrations of pulegone and the positive control metaldehyde at different times on the 1st, 2nd, 3rd and 4th day are shown in Table 8:

[0097] Table 8

[0098]

[0099] As can be seen from Table 8, pulegone and metaldehyde have different degrees of snail killing activity on the striped snail. With the increase of the concentration of the agent, the mortality rate of the striped snail gradually increased, and the corrected mortality rate of the striped snail increased with the increase of the concentration of the test solution. The longer the time, the higher the corrected mortality rate. On the first day, the corrected mortality rates of pulegone and metaldehyde on the striped snail were 100% and 33%, respectively. On the second day, the corrected mortality rates of pulegone and metaldehyde on the striped snail were 100% and 70%, respectively. On the third day, the corrected mortality rates of pulegone and metaldehyde on the striped snail were 100% and 90%, respectively. On the fourth day, the corrected mortality rate of metaldehyde was 96%. The corrected mortality rate of pulegone was over 90%.

[0100] The stomach toxicity of different concentrations of pulegone and the positive control metaldehyde to the snails at different times of 1, 2, 3 and 4 days is shown in Table 9:

[0101] Table 9

[0102]

[0103] From Table 9, we can conclude that at the first day, the LC values ​​of pulegone and metaldehyde were 50 The LC values ​​of pulegone, tea saponin and metaldehyde were 8.74 and 142.51 mg / mL, respectively. 50 On the third day, the LC50 of pulegone was 6.59 mg / mL, and the LC50 of metaldehyde was 50 was 0.83 mg / mL. On the 4th day, the LC values ​​of metaldehyde and pulegone 50 The values ​​were 0.27 and 6.45 mg / mL.

[0104] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of pulegone in pest control, characterized in that: Pulegone is used to kill harmful organisms or destroy the reproduction and propagation of harmful organisms, and the harmful organisms are golden apple snails.

2. The use according to claim 1, characterized in that: The LC values ​​of pulegone on Pomacea canaliculata at 24 h, 48 h, 72 h, 96 h, 120 h and 144 h 50 They are 542.00 mg / L, 182.23 mg / L, 55.88 mg / L, 49.45 mg / L, 45.83 mg / L and 43.38 mg / L respectively.

3. The use according to claim 1, characterized in that: The specific way in which the pulegone is used to destroy the reproduction and propagation of harmful organisms is to inhibit the spawning of golden apple snails or inhibit the hatching of golden apple snail eggs.

4. The use according to claim 3, characterized in that: The concentration of pulegone that inhibits the spawning of Pomacea canaliculata snails is 20-40 mg / L, and the treatment time is more than 2 h.

5. The use according to claim 4, characterized in that: The concentration of pulegone that inhibits the hatching of Pomacea canaliculata eggs is 30-60 mg / L, and the treatment method is direct spraying.

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