Dimethyl phthalate black fly repellent, preparation method and application thereof
Patent Information
- Application Number
- CN202311373285.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-20
- Publication Date
- 2026-10-30
- Estimated Expiration
- 2043-10-20
AI Technical Summary
[0003]目前蝇蛆病的防治主要依靠农药，药物驱虫治疗，除了杀死寄生部位的蝇蛆，对环境中的黑须污蝇起不到作用，等药效过后，继续产蛆、反复感染
[0014]本发明公开了一种邻苯二甲酸二甲酯黑须污蝇驱避剂、制备方法及其应用，在本发明中通过气相色谱-触角电位联用技术(GC-EAD)、触角电位技术(EAG)、行为实验测定黑须污蝇信息素—邻苯二甲酸二甲酯的触角电位和行为反应，了解邻苯二甲酸二甲酯对黑须污蝇行为的影响。通过实验证实，使用浓度分别为10-4μg/μL、10-3μg/μL、10-2μg/μL或10-1μg/μL的一种邻苯二甲酸二甲酯黑须污蝇驱避剂可以驱避黑须污蝇，从而减少黑须污蝇与双峰驼的接触机会，可有效防治黑须污蝇引起的阴道蝇蛆病。
Smart Images
Figure CN117617249B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biological control technology, specifically relating to a dimethyl phthalate repellent for black-bearded flies, its preparation method, and its application. Background Technology
[0002] The black-whiskered fly (Wohlfahrtia magnifica), also known as the flesh fly, belongs to the genus *Wohlfahrtia* of the subfamily Paramacronychiinae in the family Sarcophagidae of the order Diptera. It is widely distributed in the Mongolian Plateau, the Mediterranean Basin, Eastern Europe, Central Europe, North Africa, Central Asia, and North Asia. The black-whiskered fly prefers warmth and light, and is mainly active in summer. It is a major pathogen causing bleeding wounds and myiasis in natural orifices of livestock (horses, camels, dogs, goats, sheep, cattle, etc.), and is also the sole pathogen causing vaginal myiasis in Bactrian camels. In addition, the larvae of the black-whiskered fly can also cause various diseases of the ears, eyes, skin, and vulva in humans.
[0003] Currently, the control of fly myiasis mainly relies on pesticides. While pesticide treatment kills the maggots in the parasitic parts, it has no effect on the black-haired flies in the environment. Once the pesticide's effect wears off, the flies continue to produce maggots, leading to repeated infections. To change this passive control method based primarily on pesticides, we need to explore proactive biological control measures. Summary of the Invention
[0004] The purpose of this invention is to solve the existing technical problems and provide a dimethyl phthalate repellent for black-bearded flies, its preparation method and application. This repellent can repel adult black-bearded flies, thereby reducing their contact with the host and affecting their mating and maggot-producing behavior, providing a new approach for the treatment of vaginal myiasis in Bactrian camels.
[0005] This invention is achieved through the following technical solution:
[0006] A dimethyl phthalate repellent for black-bearded flies is prepared as follows: dimethyl phthalate is mixed with a solvent to form a solution.
[0007] In the above technical solution, the solvent is n-hexane.
[0008] In the above technical solution, the concentration of dimethyl phthalate is 10. -4 μg / μL to 10 -1 μg / μL.
[0009] In the above technical solution, the concentration of dimethyl phthalate is preferably 10. -4μg / μL, 10 -3 μg / μL, 10 -2 μg / μL or 10 -1 μg / μL.
[0010] Application of a dimethyl phthalate repellent for black-bearded flies in the prevention and control of livestock diseases: The above repellent is used to repel adult black-bearded flies, thereby reducing their contact with the host and affecting their mating and maggot-producing behavior.
[0011] In the above technical solution, the livestock are horses, camels, dogs, goats, sheep, cattle, etc.
[0012] In the above technical solution, the patient suffers from bleeding trauma and / or natural pore myiasis.
[0013] The beneficial effects of this invention are:
[0014] This invention discloses a dimethyl phthalate (DMT) repellent for black-bearded flies, its preparation method, and its applications. In this invention, the antennal potential and behavioral response of the black-bearded fly pheromone—dimethyl phthalate—were measured using gas chromatography-electrophoresis-electrode arthrometry (GC-EAD), antennal potential arthrometry (EAG), and behavioral experiments to understand the effect of DMT on the behavior of black-bearded flies. Experiments confirmed that using a concentration of 10... -4 μg / μL, 10 -3 μg / μL, 10 -2 μg / μL or 10 -1 A dimethyl phthalate repellent for black-bearded flies (μg / μL) can repel black-bearded flies, thereby reducing the chance of contact between black-bearded flies and Bactrian camels, and can effectively prevent vaginal myiasis caused by black-bearded flies. Attached Figure Description
[0015] Figure 1 This image shows the GC-EAD reaction of *Hemiberlesia lataniae* to dimethyl phthalate. Note: The left image shows the GC-EAD reaction of female *Hemiberlesia lataniae* to dimethyl phthalate; the right image shows the GC-EAD reaction of male *Hemiberlesia lataniae* to dimethyl phthalate. In the images, FID represents the mass spectrum of the compound, EAD represents the antennal reaction of *Hemiberlesia lataniae* to the compound, and the three EADs represent three replicate experiments of antennal potential.
[0016] Figure 2 The EAG reaction of black-bearded flies to different concentrations of dimethyl phthalate.
[0017] Figure 3 Comparison of the EAG reaction of black-bearded flies to dimethyl phthalate.
[0018] Figure 4is the behavioral response of female Wohlfahrtia magnifica to different concentrations of dimethyl phthalate (female).
[0019] Figure 5 is the behavioral response of Wohlfahrtia magnifica to different concentrations of dimethyl phthalate (male).
[0020] Figure 6 is the comparison of behavioral responses of Wohlfahrtia magnifica to dimethyl phthalate.
[0021] Note: ns: no significant difference (P>0.05) *: significant difference (0.01<P<0.05) **: extremely significant difference (P<0.01). For a person of ordinary skill in the art, on the premise of no creative work, other related drawings can be obtained from the above accompanying drawings. DETAILED DESCRIPTION
[0022] In order to make the technical means, creative features, objectives and effects of the present invention easy to understand, the technical solution of the present invention will be further described below in conjunction with specific examples.
[0023] Example 1: GC-EAD response of Wohlfahrtia magnifica to dimethyl phthalate
[0024] (1) Sample preparation: The purchased dimethyl phthalate (99%) is prepared to 10 with HPLC-grade n-hexane -2 μg / μL of test sample, and stored in a -20°C refrigerator for later use.
[0025] (2) Preparation of capillary tube: Use an alcohol lamp to soften the capillary tube, manually pull the capillary tube until the tip is just large enough to fit the insect antenna, and keep the process as balanced and stable as possible. After the capillary tube is prepared, it is filled with a conductive liquid, and 0.9% normal saline is filled in this experiment.
[0026] (3) Antenna connection:
[0027] In the experiment, female and male Wohlfahrtia magnifica of about 7 days old were selected to test dimethyl phthalate at a concentration of 10 -2 μg / μL. Each sample was tested 3 times for each of female and male Wohlfahrtia magnifica, and each antenna was used only once. Under a microscope, the antenna of Wohlfahrtia magnifica was cut off from the base, and both ends of the antenna were connected to the two glass electrodes of the electroantennograph using glass capillaries containing normal saline. The base of the antenna was connected to the reference electrode, and the top was connected to the recording electrode. After the EAD baseline of the electroantennograph was stable, sample injection was started and the recording electrode was recorded. The recorded signal is amplified and converted by the Syntech IDAC-2 signal collector, and then data acquisition and analysis are performed through GC-EAD software.
[0028] (4) Gas chromatography (GC) conditions:
[0029] The initial temperature was 50℃ and held for 1 min. The temperature was then increased to 120℃ at a rate of 5℃ / min and held for 1 min. The temperature was then increased to 150℃ at a rate of 10℃ / min and held for 1 min. Finally, the temperature was increased to 280℃ at a rate of 20℃ / min and held for 10 min. The injection volume was 1 μL. The injection was splitless.
[0030] Example 2: EAG reaction of black-bearded flies with dimethyl phthalate
[0031] (1) Sample preparation: Dimethyl phthalate was prepared into a solution of 10 μL using chromatographic grade n-hexane. -4 μg / μL, 10 -3 μg / μL, 10 -2 μg / μL and 10 -1 The test sample of μg / μL should be stored at -20℃ for later use.
[0032] (2) Check if the level of distilled water in the gas washing bottle is half full, and add more if necessary.
[0033] (3) Turn on the power supply of the CS-55 stimulation airflow generator and the relevant software on the computer.
[0034] (4) Prepare the antennae of female and male black-bearded flies that are about 7 days old and attach the antennae.
[0035] (5) Drop 10 μL of the sample to be tested evenly onto the filter paper strip, place the filter paper strip into the blue gun tip, and stimulate it after the solvent evaporates for 5 minutes. Use n-hexane as a blank control. After testing each antenna, perform the EAG measurement of the blank control to correct the EAG reaction value of the sample.
[0036] (6) Stimulation conditions: stimulation time is 0.2s, stimulation interval is 60s, stimulation airflow rate is 300mL / min, and continuous airflow is set to 1000mL / min.
[0037] (7) Each sample was measured three times to distinguish between male and female.
[0038] (8) The EAG measurement results are recorded by the EAG signal recording and analysis program. The calculation method for the relative response value of the sample antennal potential (EAG relative response value) is as follows:
[0039] EAG relative response value = (2 × antennal potential response value of the test sample) ÷ (antennal potential response value of the solvent control before the test sample + antennal potential response value of the solvent control after the test sample)
[0040] (9) Experimental results: One-way ANOVA and t-test were performed on the data using GraghPad Prism 8.0.1 and plotted.
[0041] Example 3: Behavioral response of black-bearded flies to dimethyl phthalate
[0042] (1) Connect the following devices sequentially using a water purifier tube: atmospheric sampler (outlet) → drying tower containing activated carbon → humidification device → odor bottle → Y-shaped test tube. Maintain the airflow in both arms at 1 L / min. Before the experiment, all instruments should be cleaned three times each with distilled water and alcohol to reduce errors. Select black-bearded flies around 5 days old for behavioral testing. Place only one black-bearded fly at the straight arm of the Y-shaped test tube each time, with a reaction time of 5 min. Record the result when the fly enters more than 1 / 2 of the side arm and stays for at least 30 seconds. If no selection is made within 5 min, stop the behavioral test for that fly and do not record it. Use 20 female and 20 male black-bearded flies in each group, repeat the test 3 times, and test each fly only once at the same concentration. Use distilled water as a control. The experiment is conducted at room temperature.
[0043] (2) The experimental data were analyzed and plotted using the GraghPad Prism test. If the response of black-bearded flies to the compound was significantly higher than that to the control group, then the compound had an attractive effect on black-bearded flies; conversely, if the response of black-bearded flies to the control group was significantly higher than that to the compound, then the compound had a repellent effect on black-bearded flies; if the responses of black-bearded flies to both were not significant, then it could not be determined whether the compound had an attractive or repellent effect on black-bearded flies.
[0044] Table 1. Relative EAG response values of black-bearded flies to different concentrations of dimethyl phthalate.
[0045]
[0046]
[0047] Results Analysis
[0048] Depend on Figure 1 It can be seen that adult black-bearded flies of different ages have different effects on 10 -2 Dimethyl phthalate at μg / μL all showed GC-EAD reaction.
[0049] Table 1 shows the results of different concentrations of dimethyl phthalate (EAG) measured by female black-bearded flies. Figure 2(Left figure) As shown, the experimental results showed that female black-bearded flies exhibited antennal responses to different concentrations of dimethyl phthalate. As the concentration of dimethyl phthalate increased, the relative EAG response value decreased. There was no significant difference in the relative EAG response value of female black-bearded flies to different concentrations of dimethyl phthalate (P>0.05).
[0050] Table 1 shows the results of EAG assays on different concentrations of dimethyl phthalate (DMT) in male black-whiskered flies. Figure 2 (See right figure). The experimental results show that different concentrations of dimethyl phthalate can induce antennal responses in male black-bearded flies. As the concentration of dimethyl phthalate increases, the relative EAG response value shows a trend of first increasing, then decreasing, and then increasing again. There is no significant difference in the relative EAG response value of male black-bearded flies to different concentrations of dimethyl phthalate (P>0.05).
[0051] Comparison of EAG (dimethyl phthalate) assay results between different sexes of black-bearded flies: Figure 3 As shown in the figure. The results showed that there was no significant difference in the EAG assay results of dimethyl phthalate between female and male black-bearded flies (P > 0.05).
[0052] The behavioral responses of female black-bearded flies to different concentrations of dimethyl phthalate are as follows: Figure 4 As shown. When the concentration of dimethyl phthalate is 10... -1 At a concentration of μg / μL, it showed significant repellency against female black-bearded flies (0.01 < P < 0.05); other concentrations did not show significant behavioral response against female black-bearded flies (P > 0.05).
[0053] The behavioral responses of male black-whiskered flies to different concentrations of dimethyl phthalate are as follows: Figure 5 As shown in the figure. The results indicate a concentration of 10. -4 μg / μL, 10 -3 μg / μL and 10 -2 Dimethyl phthalate at μg / μL had no significant behavioral response in male black-whiskered flies (P>0.05); while 10 μg / μL... -1 Dimethyl phthalate at a concentration of μg / μL exhibits a highly significant repellency against male black-bearded flies (P < 0.01).
[0054] Behavioral responses of different sexes of black-bearded flies to dimethyl phthalate (DMT) are as follows: Figure 6 As shown in the figure. The results showed that different concentrations of dimethyl phthalate had no significant attraction or repellency to either female or male black-bearded flies (P > 0.05).
[0055] The present invention has been described above by way of example. It should be noted that any simple modifications, alterations or other equivalent substitutions that can be made by those skilled in the art without creative effort without departing from the core of the present invention fall within the protection scope of the present invention.
Claims
1. The use of a dimethyl phthalate repellent for black-bearded flies in the preparation of drugs for bleeding wounds or natural canal myiasis, characterized in that, The repellent is prepared by the following method: dimethyl phthalate is mixed with a solvent to form a solution, wherein the concentration of dimethyl phthalate is 10. -4 μg / μL to 10 -1 μg / μL.
2. The application according to claim 1, characterized in that: The solvent is n-hexane.
3. The application according to claim 1, characterized in that: The concentration of dimethyl phthalate is 10. -4 μg / μL, 10 -3 μg / μL, 10 -2 μg / μL or 10 -1 μg / μL.