Mixing agent for repelling ant pests and application thereof

Abstract

The invention relates to a mixing agent for repelling ant pests and application thereof. The mixing agent of diethyl glutarate, 3-methylindole and 2-(formamido) benzoic acid, which is screened and developed from migratory locust reflux liquid, has a relatively good repelling effect on ants, especially solenopsis invicta. Compared with a chemical repellent agent used in the prior art, natural insect defense substances are adopted, higher environmental protection property and safety are achieved, and remarkable market value and wide popularization prospects are achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of ant pest control, and particularly relates to a mixture for repelling ant pests and its application. Background Art

[0002] Ants are an important type of pest in the human living environment. In particular, the red imported fire ant (Solenopsis invicta) is highly aggressive and poses a significant threat to children, the elderly, and individuals with allergic constitutions. As an invasive pest, the red imported fire ant is an important harmful organism to biodiversity protection and agricultural production. They invade housing, schools, lawns, etc., and have a high chance of coming into contact with humans, and stinging incidents occur frequently. In addition, these ants bite electric wires, often causing short circuits and even small fires.

[0003] Many animals and insects have developed various defense mechanisms, such as repellent substances, warning signals, cryptic colors, and toxins, to avoid predation. In addition, many insects avoid predation by releasing chemical substances accumulated from plants or secreted from their bodies. This chemical defense strategy usually involves the mixing and release of specific compounds. Therefore, the development and application of insect chemical defense substances have very important value in aspects such as biological repellents. However, there is no report on using insect regurgitant fluid to design biological agents for repelling other pests.

[0004] The herbivorous insect Locusta migratoria can produce defensive regurgitant fluid when attacked by natural enemies (ants, mantises, birds) or stimulated by the outside world. These regurgitant fluids contain special chemical substances, which have an obvious effect on repelling predators. By analyzing the chemical defense substances synthesized by these insects themselves, key components are found, which have important value in developing green and safe pest repellents and biological pesticides, etc. Summary of the Invention

[0005] The purpose of the present invention is to provide a mixture for repelling pests and its application, which can have a good repelling effect on ants, especially the red imported fire ant;

[0006] To achieve the above purpose, the technical solution adopted by the present invention is:

[0007] A mixture for repelling ant pests, the mixture includes diethyl glutarate, 3-methylindole, and 2-(formylamino)benzoic acid, wherein the concentration of diethyl glutarate is greater than 18.55 ng / μL, the concentration of 3-methylindole is greater than 0.873 ng / μL, and the concentration of 2-(formylamino)benzoic acid is greater than 0.417 ng / μL.

[0008] Another aspect of the present invention provides the application of the above-mentioned mixture in the preparation of an ant pest repellent.

[0009] Preferably, according to the above application, the solvent of the ant pest repellent is anhydrous ethanol.

[0010] On the other hand, the present invention provides an application of locust regurgitant fluid in repelling ant pests.

[0011] The present invention applies a mixture of diethyl glutarate, 3-methylindole and 2-(formylamino)benzoic acid to the place where ants are to be repelled to repel ants and red imported fire ants, and the application method can be spraying, perfusion, etc.

[0012] The mixture of diethyl glutarate, 3-methylindole and 2-(formylamino)benzoic acid screened and developed from locust regurgitant fluid by the present invention has a good repellent effect on ants and red imported fire ants. Compared with the chemical repellent agents used in the prior art, the present invention uses natural insect defense substances, has higher environmental protection and safety, and has significant market value and broad promotion prospects. Description of the Drawings

[0013] Figure 1 It is the predation relationship between locusts and red imported fire ants; among them, (a) the locust secretes regurgitant fluid when attacked by red imported fire ants; (b) the survival curve of locusts under the attack of red imported fire ants.

[0014] Figure 2 It is the LC-MS substance classification of locust regurgitant fluid.

[0015] Figure 3It is the repellent effect of potential defensive compounds in the regurgitant fluid of locusts on Solenopsis invicta; among them, (a) Schematic diagram of the experimental design; (b) Repellent results. The letters in the figure are the abbreviations of different compounds, namely: DL-Indole-3-lactic acid (DILA), Betaine (BE), 2-(Formylamino)Benzoic Acid (FBA), N-Methyl-α-aminoisobutyric acid (MA), 2-Hydroxycinnamate (HY), Choline Hydroxide (CH), Diethyl glutarate (DG), Phloretate (PH), Pipecolic acid (PA), Choline (CHO), Choline Glycerophosphate (CG), 3,4-Dimethoxyphenethylamine (DMPEA), 6-Aminohexanoate (AM), Nonadecanoic acid (NA), Benzamidine (BA), Phosphocholine (PC), Uric Acid (UA), Kynurenic acid (KA), D-Galactonic acid (GA), Trigonelline (TR), Isobutyrylglycine (IG), Phthalate (PHT), L-Malate (MAL), Stearic acid (SA), L-Carnitine (CA), 3-Methylindole (3MI), trans-Cinnamic acid (TCA), Dl-Tropic acid (TA), Glycerol-3-phosphate (GP), 2-Pyrrolidinone (PY), Maleic Acid (MAA).

[0016] Figure 4 It is the behavioral trajectory of Solenopsis invicta in the experimental area treated with potential defensive compounds in the regurgitant fluid of locusts. It can be seen that Solenopsis invicta is always avoiding the regurgitant fluid.

[0017] Figure 5 is the actual concentration of potential defensive compounds in the regurgitant fluid of Locusta migratoria.

[0018] Figure 6 is the repellent effect of potential defensive compounds and their mixtures in the regurgitant fluid of Locusta migratoria on Solenopsis invicta. Detailed implementation manners

[0019] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0020] A mixture for repelling ant pests, the mixture comprising diethyl glutarate, 3-methylindole and 2-(formylamino)benzoic acid, wherein the concentration of diethyl glutarate is greater than 18.55 ng / μL, the concentration of 3-methylindole is greater than 0.873 ng / μL, and the concentration of 2-(formylamino)benzoic acid is greater than 0.417 ng / μL.

[0021] On the other hand, the present invention provides an application of the regurgitant fluid of Locusta migratoria in repelling ant pests.

[0022] As Figure 1 shown, a is the regurgitant fluid secreted by Locusta migratoria when attacked by Solenopsis invicta. From Figure 1 b, it can be seen that the regurgitant fluid secreted by Locusta migratoria can repel Solenopsis invicta. The following experiments are carried out on the regurgitant fluid secreted by Locusta migratoria when attacked by Solenopsis invicta.

[0023] I. LC-MS separation and identification of compounds contained in the regurgitant fluid of Locusta migratoria

[0024] 1. Extraction of regurgitant fluid by artificial stimulation method

[0025] Use the index finger and thumb of one hand to clamp Locusta migratoria, and gently press its head or chest to stimulate it to secrete regurgitant fluid. Subsequently, use a pipette to collect the regurgitant fluid on the mouthparts. The obtained regurgitant fluid sample is collected into a 1.8 mL cryopreservation tube, and the regurgitant fluid of every 6 Locusta migratoria is placed in one cryopreservation tube. During collection, the cryopreservation tube is placed on ice throughout the process. After collection, all the regurgitant fluid is stored in a ultra-low temperature refrigerator at -80 °C for standby.

[0026] 2. Identification of compounds

[0027] Use a liquid chromatography-mass spectrometry (model: Exion LC-SCIEX 6500+) and adopt the multiple reaction monitoring mode (MRM) to accurately determine the samples extracted in S1. The specific analysis conditions are as follows:

[0028] (1) Chromatographic conditions: Chromatographic column: Xselect HSS T3, 2.5μm, 2.1×150mm; Mobile phase: A: 0.1% formic acid - water, B: 0.1% formic acid - acetonitrile; Column temperature: 50°C; Flow rate: 0.4 mL / min;

[0029] (2) Gradient program: 0 min: 98% A, 2% B; 2 min: 98% A, 2% B; 15 min: 0% A, 100% B; 17 min: 0% A, 100% B; 17.1 min: 98% A, 2% B; 20 min: 98% A, 2% B;

[0030] (3) Mass spectrometry conditions: 1) Positive ion mode: Curtain Gas: 35 psi; Collision Gas: Medium; IonSpray Voltage: 5500 V; Temperature: 550°C; Ion Source Gas 1: 60; Ion Source Gas2: 60; 2) Negative ion mode: Curtain Gas: 35 psi; Collision Gas: Medium; IonSpray Voltage: -4500 V; Temperature: 550°C; Ion Source Gas 1: 60; Ion Source Gas 2: 60.

[0031] The analysis results showed that 907 organic compounds were detected in the reflux fluid extract, and the compound classification is as Figure 2 shown. After sorting by peak area, 31 potential defensive compounds were screened out among the top 100 compounds before sorting.

[0032] II. Screening of potential defensive compounds in the reflux fluid of Locusta migratoria

[0033] The EthoVision XT 15.0 software of the video tracking system was used to record the repellent behavior characteristics of the potential defensive compounds in the reflux fluid of Locusta migratoria against the worker ants of Solenopsis invicta. The behavior determination was carried out in a rectangular arena (184 mm × 92 mm × 30 mm) with the side walls coated with Teflon, allowing the worker ants of Solenopsis invicta to move on the bottom. The worker ants of Solenopsis invicta were starved for 24 - 48 h before the experiment, and only water was provided during this period. A piece of mealworm about 2 mm long was completely immersed in the test compound solution (high concentration 10000 ng / μL) for at least 30 s, and then quickly fixed at the bottom of the arena ( Figure 3a). Subsequently, a video camera was used to record the activity trajectories of red imported fire ants in the arena for 10 min. The control group was an absolute ethanol solvent, and there were no less than 28 replicates for each compound. According to the recorded videos, the cumulative time of red imported fire ants foraging was manually measured using a video tracking system (EthoVision XT 15.0), and the difference in the cumulative foraging time of different treatments was compared for significance using GraphPad prism 10.1 (P = 0.05).

[0034] The results are as Figure 3 and Figure 4 shown. The results showed that there were 6 potential defensive compounds in the regurgitant fluid. The time for red imported fire ants to forage on mealworms in the control group was significantly higher than that on mealworms treated with compounds (P < 0.05), while other substances showed no trend of repellent effect on red imported fire ants (P > 0.05). Figure 3 b).

[0035] III. Determination of the actual contents of 6 potential defensive compounds in the regurgitant fluid

[0036] An HPLC-MS / MS system (HPLC: ExionLC TM AC; MS: Scienx Triple Quadruple 4500) was used to analyze the samples.

[0037] HPLC conditions: Column: HYPERSIL GOLD C18 column (3 μm, 2.1 mm * 100 mm); Mobile phase A was H2O (0.1% FA); Mobile phase B was acetonitrile. The gradient program of the mobile phase was as follows: 90% A from 0 min to 0.2 min, 90% A for 3 min and held for 8 min, 10% A for 8.1 min and held for 10 min. The flow rate was 0.3 mL / min, and the column temperature was 35 °C.

[0038] MS conditions: The triple quadrupole 4500 HPLC-MS / MS system equipped with an ESI ion source was operated in positive and negative ion modes and controlled by Analyst 1.6.3 software (Sciex). The operation parameters of the ESI source were as follows: The ion spray voltage was 5500 V / 4500 V, the source temperature was 550 °C; CUR was set at 30 psi; CAD was 9 psi.

[0039] The results are as Figure 5As shown in Table 1. The results showed that the absolute contents of the six compounds in the regurgitant fluid were as follows: DG: 18.55 ± 0.405 ng / μL; 3MI: 0.873 ± 0.0245 ng / μL; FBA: 0.417 ± 0.0857 ng / μL; DMPEA: 10.17 ± 0.381 ng / μL; BA: 0.458 ± 0.0298 ng / μL; MAA: 0.057 ± 0.0015 ng / μL.

[0040] Table 1 Potential defensive compounds and actual concentrations in the regurgitant fluid of Locusta migratoria

[0041]

[0042] IV. Repellent test of the actual concentration of defensive compounds and their mixtures in the regurgitant fluid against Solenopsis invicta

[0043] According to the actual concentrations of the six potential defensive compounds in the regurgitant fluid, the repellent behavioral characteristics of single compounds and mixtures against Solenopsis invicta were detected.

[0044] The results are as Figure 6 shown. The results showed that the mixture of diethyl glutarate, 3-methylindole and 2-(formylamino)benzoic acid had a stronger repellent effect on Solenopsis invicta ( Figure 6 ), and the foraging number and foraging time of the red imported fire ants in the control group were significantly higher than those in the foraging compound treatment group (**P < 0.01), while single compounds and other mixtures showed no tendency to repel red imported fire ants (P > 0.05).

Claims

1. A mixture for repelling ant pests, characterized in that, The mixture includes diethyl glutarate, 3-methylindole, and 2-(formylamino)benzoic acid, wherein the concentration of diethyl glutarate is greater than 18.55 ng / μL, the concentration of 3-methylindole is greater than 0.873 ng / μL, and the concentration of 2-(formylamino)benzoic acid is greater than 0.417 ng / μL.

2. Use of the mixture according to claim 1 in the preparation of an ant pest repellent.

3. The application according to claim 3, wherein The solvent of the ant pest repellent is absolute ethanol.

4. Use of the regurgitant fluid of Locusta migratoria in repelling ant pests.

Images