Ant repellent mixture and use thereof

Abstract

The present application relates to a kind of mixed agent for repelling ant pests and its application, the mixed agent of glutaric acid diethyl ester, 3-methylindole and 2-(formylamino) benzoic acid screened and developed from locust reverse flow liquid has good repelling effect to ant, especially red imported fire ant.Compared with the chemical repellent used in prior art, the present application uses insect natural defense material, has higher environmental protection and safety, has significant market value and broad popularization prospect.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of ant pest control, and particularly relates to a mixed agent for repelling ants and application thereof. BACKGROUND

[0002] Ants are an important class of pests in human living environment, especially the red imported fire ant (Solenopsis invicta) which is extremely aggressive and poses a significant threat to children, the elderly and people with allergic constitution. As an invasive pest, the red imported fire ant is an important harmful organism for biodiversity protection and agricultural production. They invade houses, schools, lawns and other places, and have more opportunities to contact with people, and the phenomenon of biting occurs from time to time. In addition, these ants gnaw on 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 coloration and toxins, to escape from predation. In addition, many insects escape from predation by releasing chemicals accumulated from plants or secreted from the body, and 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 biological repellents and other aspects. However, there is no report on the use of insect regurgitant to design biological agents to repel other pests.

[0004] The phytophagous insect locust (Locusta migratoria) can produce defensive regurgitant containing special chemicals when attacked by natural enemies (ants, mantids, birds) or external stimuli, which has obvious effect on repelling predators. Through the analysis of these insect-synthesized chemical defense substances, it is found that the key ingredients have important value in the development of green and safe pest repellents and biological pesticides. SUMMARY

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

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

[0007] A mixed agent for repelling ant pests, the mixed agent 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.

[0008] In another aspect of the present application, the application of the above-mentioned mixed agent in the preparation of an ant pest repellent is provided.

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

[0010] In another aspect, the present invention provides the application of locust reversal liquid in repelling ant-like pests.

[0011] This invention involves applying a mixture of diethyl glutarate, 3-methylindole, and 2-(formamide)benzoic acid to areas where ants are to be repelled, thereby repelling ants and red imported fire ants. The application method can be spraying, pouring, etc.

[0012] This invention discloses a mixture of diethyl glutarate, 3-methylindole, and 2-(formamide)benzoic acid, screened and developed from locust reflux sap, which exhibits good repellent effects against ants and red imported fire ants. Compared with existing chemical repellents, this invention utilizes natural insect defense substances, resulting in higher environmental friendliness and safety, significant market value, and broad prospects for promotion. Attached Figure Description

[0013] Figure 1 The predator-prey relationship between locusts and red imported fire ants is shown; (a) locusts secrete reflux fluid when attacked by red imported fire ants; and (b) the survival curve of locusts under the attack of red imported fire ants.

[0014] Figure 2 This is the LC-MS material classification of the locust reflux fluid.

[0015] Figure 3This study investigated the repellent effect of potential defensive compounds from the backflow of locusts on red imported fire ants. (a) shows the experimental design; (b) shows the repellency results. The letters in the figures are abbreviations for different compounds: DL-indole-3-lactic acid (DILA), betaine (BE), 2-(formylamino)benzoic acid (FBA), N-methyl-a-aminoisobutyric acid (MA), 2-hydroxycinnamate (HY), choline hydroxide (CH), diethyl glutarate (DG), phloretate (PH), pipecolic acid (PA), choline (CHO), and glycerophospholipid choline (CHO). 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 The following are listed: acid (TCA), tropic acid (TA), glycerol-3-phosphate (GP), 2-pyrrolidinone (PY), and maleic acid (MAA).

[0016] Figure 4 The image shows the behavioral patterns of red imported fire ants in the experimental area treated with a potential defense compound in the reflux liquid of locusts. It can be seen that the red imported fire ants are always avoiding the reflux liquid.

[0017] Figure 5 This is the actual concentration of potential defensive compounds in the locust reflux fluid.

[0018] Figure 6 It is the repellent effect of potential defensive compounds and their mixtures in the locust reflux fluid on red imported fire ants. Detailed Implementation

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

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

[0021] In another aspect, the present invention provides the application of locust reversal liquid in repelling ant-like pests.

[0022] like Figure 1 As shown, 'a' represents the reflux fluid secreted by the locust when attacked by red imported fire ants. Figure 1 b shows that the reflux secreted by locusts can repel red imported fire ants. The following experiment investigates the reflux secreted by locusts when attacked by red imported fire ants.

[0023] I. LC-MS Separation and Identification of Compounds in Locust Reflux Fluid

[0024] 1. Extraction of reflux liquid using artificial stimulation method

[0025] Hold the locust between your index finger and thumb and gently press its head or thorax to stimulate the secretion of reflux fluid. Then, collect the reflux fluid from its mouthparts using a pipette. Collect the reflux fluid samples into 1.8 mL cryovials, with reflux fluid from six locusts placed in each cryovial. Keep the cryovials on ice throughout the collection process. After collection, store all reflux fluid in an ultra-low temperature freezer at -80°C for later use.

[0026] 2. Identification of compounds

[0027] Using a liquid chromatography-mass spectrometry system (model: Exion LC-SCIEX) (6500+) and used Multiple Reaction Monitoring (MRM) mode to perform precise qualitative analysis on the samples extracted from S1. The specific analytical conditions are as follows:

[0028] (1) Chromatographic conditions: 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℃; Flow rate: 0.4mL / 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: 35psi; Collision Gas: Medium; IonSpray Voltage: 5500V; Temperature: 550℃; Ion Source Gas 1: 60; Ion Source Gas 2: 60; 2) Negative ion mode: Curtain Gas: 35psi; Collision Gas: Medium; IonSpray Voltage: -4500V; Temperature: 550℃; Ion Source Gas 1: 60; Ion Source Gas 2: 60.

[0031] The analysis results showed that 907 organic compounds were detected in the reflux extract, and the compounds were classified as follows: Figure 2 As shown in the figure. After sorting by peak area, 31 potential defensive compounds were screened from the top 100 compounds.

[0032] II. Screening of potential defense compounds for locust reflux fluid

[0033] The repellent behavior of locust backflow sap potential defense compounds on red imported fire ant workers was recorded using the EthoVision XT 15.0 video tracking system. Behavioral measurements were conducted in a rectangular arena (184 mm × 92 mm × 30 mm) with Teflon-coated sidewalls, allowing the red imported fire ant workers to move at the bottom. The red imported fire ant workers were starved for 24–48 hours prior to the experiment, during which time only water was provided. A 2 mm segment of mealworm was completely immersed in the test compound solution (high concentration 10000 ng / μL) for at least 30 seconds, and then quickly fixed to the bottom of the arena. Figure 3a) Subsequently, the activity trajectory of red imported fire ants in the arena was recorded for 10 minutes using a camcorder. The control group was anhydrous ethanol solvent, with at least 28 replicates for each compound. Based on the recorded video, the cumulative foraging time of red imported fire ants was manually recorded using a video tracking system (EthoVision XT 15.0), and the differences in cumulative foraging time between different treatments were compared using GraphPad Prism 10.1 (P = 0.05).

[0034] The results are as follows Figure 3 and Figure 4 As shown. The results indicated the presence of six potential defensive compounds in the reflux solution. Red imported fire ants spent significantly more time foraging for mealworms in the control group than in the compound-treated group (P<0.05), while other substances showed no repellent effect on red imported fire ants (P>0.05). Figure 3 b).

[0035] III. Determination of the actual content of six potential defense compounds in the reflux fluid

[0036] Using an HPLC-MS / MS system (HPLC:ExionLC) TM AC; MS: Scienx Triple Quadruple 4500) to analyze samples.

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

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

[0039] The results are as follows Figure 5As shown in Table 1, the absolute contents of the six compounds in the reflux liquid were: 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 locust reflux fluid.

[0041]

[0042] IV. Repellency Tests of Defensive Compounds and Their Mixtures at Actual Concentrations in the Reflux Liquid against Red Imported Fire Ants

[0043] The repellent behavior of single compounds and mixtures against red imported fire ants was determined based on the actual concentrations of six potential defensive compounds in the reflux fluid.

[0044] The results are as follows Figure 6 As shown. The results indicate that the mixture of diethyl glutarate, 3-methylindole, and 2-(formamide)benzoic acid has a stronger repellent effect on red imported fire ants. Figure 6 The number of red imported fire ants foraging and the foraging time in the control group were significantly higher than those in the group treated with the foraging compound (**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-like pests, characterized in that, The mixture comprises diethyl glutarate, 3-methylindole, and 2-(formamide)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-(formamide)benzoic acid is greater than 0.417 ng / μL.

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

3. The application according to claim 2, characterized in that, The solvent for the ant pest repellent is anhydrous ethanol.

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