CN114982754A - Methylheptenone drosophila melanogaster attractant, preparation method and application thereof - Google Patents
Methylheptenone drosophila melanogaster attractant, preparation method and application thereof Download PDFInfo
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- JPTOCTSNXXKSSN-UHFFFAOYSA-N methylheptenone Chemical compound CCCC=CC(=O)CC JPTOCTSNXXKSSN-UHFFFAOYSA-N 0.000 title claims abstract description 61
- 239000005667 attractant Substances 0.000 title claims abstract description 15
- 230000031902 chemoattractant activity Effects 0.000 title claims abstract description 15
- 241000255601 Drosophila melanogaster Species 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 241001599018 Melanogaster Species 0.000 claims abstract description 39
- 241000282828 Camelus bactrianus Species 0.000 claims abstract description 9
- 208000006123 Myiasis Diseases 0.000 claims abstract description 5
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical group CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 16
- 239000002904 solvent Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 6
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01M—CATCHING, TRAPPING OR SCARING OF ANIMALS; APPARATUS FOR THE DESTRUCTION OF NOXIOUS ANIMALS OR NOXIOUS PLANTS
- A01M1/00—Stationary means for catching or killing insects
- A01M1/02—Stationary means for catching or killing insects with devices or substances, e.g. food, pheronones attracting the insects
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01M—CATCHING, TRAPPING OR SCARING OF ANIMALS; APPARATUS FOR THE DESTRUCTION OF NOXIOUS ANIMALS OR NOXIOUS PLANTS
- A01M2200/00—Kind of animal
- A01M2200/01—Insects
- A01M2200/012—Flying insects
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses a methyl heptenone drosophila melanogaster attractant, a preparation method and application thereof. According to the invention, the influence of the methylheptenone on the luring behavior of the dirofilaria melanogaster is known by measuring the tactile potential and the behavior reaction of the dirofilaria melanogaster to a bactrian camel vagina volatile substance, namely methyl heptenone, through a gas chromatography-tactile potential combined technology (GC-EAD), an tactile potential technology (EAG) and a behavior experiment. Experiments prove that the methyl heptenone used in the invention can attract the black flies, thereby reducing the contact chance of the black flies and bactrian camels, and effectively preventing and treating vaginal myiasis caused by the black flies.
Description
Technical Field
The invention belongs to the technical field of biological prevention and control, and particularly relates to a methyl heptenone Musca melanogaster attractant, a preparation method and application thereof.
Background
The dirofilaria fuscogilva belongs to Diptera (Diptera), Sarcophagidae (Sarcophagidae), Subfamily Musca (Subfamily Paranarycina), and the Genus Drosophila (Genus Wohlfahrita Brauer), which are important pathogens causing traumatic myiasis in humans and animals. The black beard maggot is the only pathogen causing bactrian camel vaginal maggot disease and is widely distributed in southern Europe, the middle east, North Africa and Xinjiang, inner Mongolia and other areas of China. The infestation of the Musca melanogaster varies from area to area, and the extent of the infestation varies from area to area, depending on the composition of the parasite fauna, the host, the animal husbandry and control practices, climate and geography related factors. In europe there are reports of a related infection rate of sheep infested with black flies of up to 90%. However, in other areas, bactrian camels are the key infesting objects of the black-spotted fly larvae. In one report by iran, it was found that the traumatic disease caused by larvae of the dirofilaria melanogaster was about 60%. Dogs were subjected to a traumatic myositis test at both farms in the northern part of morocco and all larvae and adults found and grown from them were identified as dirofuscus nigricans. Other animals have also been reported to be infected by larvae of the Musca melanogaster. In addition, the larvae of the Musca nigra can cause various diseases in human beings, such as diseases of ears, eyes, vulva, skin, and the like.
The black beard dirty flies mainly cause vaginal myiasis of female camels in China, seriously affect the growth and development performance of the sick camels and the development of camel industry, and become the primary harmful diseases of the female camels. At present, the prevention and the treatment of the fly maggot disease mainly depend on pesticides and drug-repellent treatment, except killing the fly maggots at parasitic parts, the fly maggots do not play a role in environmental black beard dirty flies, and after the pesticide effect is over, the fly maggots continue to be produced and are repeatedly infected. In order to change the current situation of pesticide-based control, people need to explore biological control measures mainly for the black bearded flies in the environment of artificial interference.
Disclosure of Invention
The invention aims to solve the technical problems in the prior art and provides a methyl heptenone dirofuscus nigricans attractant which can attract the imago of dirofuscus nigricans to reduce the contact chance between the imago and a host, thereby influencing the mating and maggot producing behaviors and providing a new idea for treating bactrian camel vaginosis.
The invention is realized by the following technical scheme:
the methyl heptenone drosophila melanogaster attractant is prepared by the following steps:
methyl heptenone is dissolved in the solvent.
In the technical scheme, the solvent is n-hexane.
In the technical scheme, the concentration of the methylheptenone is 10 -4 μ g/. mu.L to 1 μ g/. mu.L.
A preparation method of methyl heptenone dirofusmelanogaster attractant comprises the following steps:
methyl heptenone is dissolved in the solvent.
In the technical scheme, the solvent is n-hexane.
In the technical scheme, the concentration of the methylheptenone is 10 -4 μ g/. mu.L to 1 μ g/. mu.L.
The application of the methylheptenone drosophila melanogaster attractant comprises the following steps: the muscovado melanogaster is attracted by the methyl heptenone, the contact chance of the muscovado melanogaster and the bactrian camel is reduced, and the vaginal myiasis caused by the muscovado melanogaster is prevented and controlled.
The invention has the beneficial effects that:
according to the invention, the influence of the methylheptenone on the luring behavior of the dirofilaria melanogaster is known by measuring the tactile potential and the behavior reaction of the dirofilaria melanogaster to a bactrian camel vagina volatile substance, namely methyl heptenone, through a gas chromatography-tactile potential combined technology (GC-EAD), an tactile potential technology (EAG) and a behavior experiment. Experiments prove that the methyl heptenone used in the invention can attract the black flies, thereby reducing the contact chance of the black flies and bactrian camels and effectively preventing and controlling vaginal myiasis caused by the black flies.
Drawings
FIG. 1 is a GC-EAD reaction of 1 day old Musca melanogaster on methyl heptenone.
FIG. 2 is a GC-EAD reaction of 7-day-old Musca melanogaster on methyl heptenone.
FIG. 3 is the EAG response of male Musca melanogaster to varying concentrations of methylheptenone.
FIG. 4 is the EAG response of female Musca melanogaster to varying concentrations of methylheptenone.
FIG. 5 is a comparison of the EAG reaction of Musca melanogaster to methylheptenone.
FIG. 6 is a graph showing the behavioral response of male Musca melanogaster to methylheptenone.
FIG. 7 is a graph showing the behavioral response of female Drosophila melanogaster to methylheptenone.
FIG. 8 is a comparison of the behavioral response of Musca melanogaster to methylheptenone.
For a person skilled in the art, other relevant figures can be obtained from the above figures without inventive effort.
Detailed Description
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the technical scheme of the invention is further explained by combining the specific embodiments.
Examples
The electrophysiological and behavioral reaction of methyl heptenone on the dirofilaria melanogaster comprises the following steps:
GC-EAD reaction of Musca melanogaster to methyl heptenone
(1) Sample preparation: preparing the purchased methylheptenone (more than or equal to 97 percent) into 10 percent by using HPLC grade n-hexane -2 μ g/. mu.L of the sample to be tested.
(2) Preparing a glass capillary tube: firstly, automatically preparing by using an electrode drawing instrument; ② under the alcohol lamp, the drawing is manually carried out, and the drawing process is kept balanced as much as possible. The drawn capillary tip can just as well be placed into the insect antenna. After the preparation of the glass capillary tube is finished, 0.9% of normal saline is filled.
(3) Preparing and connecting antennae: the tentacles of the black-beard flies are cut off from the base part by an ophthalmic surgical scissors under a microscope, the tentacles are absorbed into a capillary by utilizing the surface tension of physiological saline in the capillary, the capillary is lapped on two glass electrodes of an tentacle potentiometer, the base part of the tentacles is connected with a reference electrode, and the top part of the tentacles is connected with a recording electrode. And after the EAD base line is basically stable, starting sample injection and recording the electrode. And the recorded signals are amplified and converted by an IDAC-2 signal collector, and then data acquisition and analysis are carried out by GC-EAD software. Selecting 1-day-old and 7-day-old black beard flies to respectively determine 10 pairs -2 GC-EAD reaction of μ g/. mu.L methylheptenone was repeated 3 times for each sample and once for each antenna.
(4) GC conditions were as follows: the initial temperature was 120 deg.C, held for 1min, and the temperature was raised to 280 deg.C at a rate of 20 deg.C/min, held for 15 min. Split-flow injection at 5: 1, and manually inject 1 μ L of sample each time.
EAG reaction of Musca melanogaster to methyl heptenone
(1) Sample preparation: preparing methyl heptenone with chromatographic grade n-hexane into 10 -4 μg/μL、10 -3 μg/μL、10 -2 μg/μL、10 -1 mu.g/mu.L and 1 mu.g/mu.L of samples to be detected, and storing the samples to be detected in a refrigerator at the temperature of-20 ℃ for later use.
(2) Selecting adult Musca melanogaster of about 5-7 days old, cutting off the tentacles from the base part, and connecting the tentacles. Taking 10 mu L of sample to be tested, uniformly dripping the sample on a filter paper strip with the thickness of 30mm multiplied by 10mm, putting the filter paper strip into a clean 1mL blue gun head, and stimulating after volatilizing the solvent for about 5 min. And (3) taking n-hexane (HPLC grade) as a blank control, and performing blank EAG measurement before and after each antenna EAG measurement to correct the EAG reaction value of the sample. Samples were tested in a random fashion and the assay was performed at room temperature. The end of the dropper is connected with a stimulating gas control device, each stimulating time is 0.2s, the stimulating interval is 60s, the flow rate of stimulating gas flow is set to be 300mL/min, and the continuous gas flow is set to be 1000 mL/min. Each sample was replicated 6 times to distinguish males from females. The sequence of EAG stimulation for each sample was: blank Control (CK) → 10 -4 μg/μL→10 -3 μg/μL→10 -2 μg/μL→10 -1 μ g/μ L → 1 μ g/μ L → blank Control (CK).
The relative reaction value of EAG is calculated as follows:
the relative value of the response of the sample antenna potential is 2 × (response of the antenna potential of the test sample)/(response of the antenna potential of the solvent control before the test sample + response of the antenna potential of the solvent control after the test sample)
(3) Statistical analysis: the EAG response data were collated using Excel 16.0, analyzed and plotted using GraphPad prism8.0.1, and the EAG responses for different test samples of the same sex and the same test sample of the same sex were analyzed using One-Way ANOVA (P < 0.05).
Behavioral reaction of Musca melanogaster to methyl heptenone
(1) The water purifier pipe is sequentially connected with an atmospheric sampling instrument (air outlet) → a drying tower filled with active carbon → a wetting device → a smell bottle → a Y-shaped test tube, and the air flow of the two side arms is maintained at 1L/min. Selecting about 5-7 days old black beard fly, and performing behavior experiment determination. Only one black beard fly is placed in the straight arm opening of the Y-shaped test tube each time, the reaction time is 5min, and when the insect enters the side arm 1/2 and stays for at least 30s, the result is taken as a selection result and recorded. If no selection is made within 5min, the behavioral determination of the insect is stopped and no record is made. The female and male worms record 60 insects respectively, 3 groups are repeated, each group has 20 insects, each insect with the same concentration is tested only once, 5 insects are tested each time, the Y-shaped test tube is horizontally rotated by 180 degrees, and the positions of two arms are exchanged to reduce errors. After testing 10 head worms, the inner arm of the Y-shaped tube is cleaned by 75% alcohol and distilled water respectively, and then placed in a drying box for drying, and a clean Y-shaped test tube is replaced. The experiment was continued. The black beard fly active period was mainly performed during the day, so the test time was 9: 00-17: 00, the entire experiment was performed at room temperature, fasted for 12 hours prior to the experiment. The test was also performed from low to high concentration with n-hexane as a blank.
(2) Counting: behavioral response data were collated using Excel 16.0, analyzed on 3 compounds using GraphPad prism8.0.1 using t-test, and plotted using Origin PRO.
Analysis of results
As can be seen from FIGS. 1 and 2, pairs of male and female imagoes of Musca melanogaster at different ages in days 10 -2 Mu.g/. mu.L of methylheptenone were subjected to GC-EAD reaction.
The relative EAG responses of male Musca melanogaster to various concentrations of methylheptenone are shown in FIG. 3. Analysis shows that the relative reaction values of EAG of male Musca melanogaster to methyl heptenone with different concentrations are different, the relative reaction value of EAG is firstly increased, then decreased and then increased along with the increase of the concentration of the methyl heptenone, and when the concentration is maximum, the relative reaction value reaches the maximum. The change of the relative reaction value of the EAG is smaller and smaller along with the increase of the concentration. 10 -4 μg/μL、10 -3 μg/μL、10 -2 μg/μL、10 -1 The relative reaction values of EAG of mu g/mu L and EAG of 1 mu g/mu L are 1.167mv, 1.313mv, 1.688mv, 1.654mv and 1.803mv respectively. Methyl heptane of different concentrationsKetene causes no significant difference in EAG relative response values of male dirofilaria melanogaster (P > 0.05).
The relative EAG response values of female dirofilaria melanogaster to different concentrations of methylheptenone are shown in fig. 4. The result shows that female dirofilaria melanogaster has certain EAG reaction to methyl heptenone with different concentrations, and the relative reaction value changes with the concentration. The relative EAG response of female Musca melanogaster increased with increasing concentration of methylheptenone. When the concentration is 1 mug/muL, the EAG relative reaction value is 2.138mv and is obviously higher than 10 mv -4 μ g/. mu.L and 10 -3 Relative reaction value of EAG at μ g/. mu.L concentration (P < 0.05), 10 -4 μg/μL、10 -3 μg/μL、10 -2 μ g/. mu.L and 10 -1 The relative reaction values of the EAG of mu g/mu L are 1.157mv, 1.263mv and 1.824mv1.841mv respectively, and the rest are not obvious (P is more than 0.05).
The EAG response pair of the different sex Drosophila melanogaster to methylheptenone is shown in FIG. 5. Analysis shows that the EAG relative reaction value of the male and female adults to the methyl heptenone is not obviously different (P is more than 0.05).
The results of the behavioral response of male Musca melanogaster to methylheptenone (FIG. 6), 10 -2 The mu g/mu L has obvious attraction to male black flies (P is less than 0.05); when the concentration is 10 -1 At the time of mu g/mu L and 1 mu g/mu L, the bait has very obvious attraction to male black flies
The male Musca melanogaster showed no significant behavioral response to low concentrations of methylheptenone (P > 0.05).
The results of the behavioral response of female Musca melanogaster to methylheptenone (FIG. 7), 10 -2 μg/μL、10 -1 Both mu g/mu L and 1 mu g/mu L of methylheptenone have very significant attractancy to female dirofuscus nigricans (P < 0.01). Female dirofilaria melanogaster had no significant behavioral response to other concentrations of methylheptenone (P > 0.05).
The behavior experiment of the different sex of Musca melanogaster on methyl heptenone (figure 8) shows that when the concentration is 10% -1 The attraction of the composition to female black-spotted flies is obviously higher than that of males (P is less than 0.0) at the concentration of mu g/mu L5) (ii) a When the concentration was 1. mu.g/. mu.L, the attractancy to the female was extremely significant compared to that of the male Musca melanogaster. The other concentrations have no significant difference (P > 0.05).
The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.
Claims (7)
1. The methyl heptenone drosophila melanogaster attractant is characterized by being prepared by the following method: methyl heptenone is dissolved in the solvent.
2. The methyl heptenone drosophila melanogaster attractant of claim 1, wherein: the solvent is n-hexane.
3. The methyl heptenone drosophila melanogaster attractant of claim 1, wherein: the concentration of the methyl heptenone is 10 -4 μ g/. mu.L to 1 μ g/. mu.L.
4. A preparation method of methyl heptenone drosophila melanogaster attractant is characterized by comprising the following steps: methyl heptenone is dissolved in a solvent.
5. The method for preparing the methyl heptenone drosophila melanogaster attractant disclosed by claim 4, wherein the method comprises the following steps: the solvent is n-hexane.
6. The method for preparing the methyl heptenone drosophila melanogaster attractant disclosed by claim 4, wherein the method comprises the following steps: the concentration of the methyl heptenone is 10 -4 μ g/. mu.L to 1 μ g/. mu.L.
7. The application of the methylheptenone drosophila melanogaster attractant is characterized in that: the melanogaster is attracted by the methyl heptenone, so that the contact chance of the melanogaster and the bactrian camel is reduced, and the biological prevention and control effect on the vaginal myiasis caused by the melanogaster is achieved.
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| CN109042663A (en) * | 2018-09-10 | 2018-12-21 | 柳州市华博有害生物防治有限公司 | Fruit fly trapping bait and its application |
| CN110720460A (en) * | 2019-11-12 | 2020-01-24 | 山西农业大学 | Plant source pollution-free active substance for attracting green gram, attractant and method |
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