Apparatus for observing the behavioral response of small insects to pesticide deposition patterns and method of use
Patent Information
- Application Number
- CN202610693675.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-05-20
- Publication Date
- 2026-08-21
AI Technical Summary
相较于传统生物测定方法，如Potter喷雾法因试虫活动范围过大、叶面药斑不易观察等原因难以明确杀虫剂雾滴的实际利用率，如采用浸叶法形成全域有药区，则无法获知试虫对有药与无药区的行为选择性，无法模拟田间的真实状态
[0017]采用上述的技术方案，本发明与现有技术相比，其具有的有益效果为：本方案装置结构简单，且操作灵活便利，本方案装置将杀虫剂沉积药斑与试虫取食区域相结合，显著提高了试虫接触药斑的几率；且通过装置迷你化可缩小试虫活动范围，提升药斑接触行为观察的成功率。本装置材料来源方便，容易定制，检测结果可靠，重复性好，非常适用于开展杀虫剂应用防治微小昆虫前的施药条件优化检测；基于该装置，通过观测微小刺吸式昆虫对杀虫剂沉积药斑的行为反应，可以为明确不同杀虫剂沉积药斑条件对微小刺吸式昆虫行为反应的影响以及48 h后准确的毒力效果提供积极可行的研究辅助，为杀虫剂采用无人机喷雾时选择适宜的雾滴粒径提供有价值的参考。
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Figure CN122603818A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the fields of insect behavior observation technology and insecticide resistance testing technology for piercing-sucking pests, and particularly to a device and method for observing the behavioral response of tiny insects to insecticide deposits. Background Technology
[0002] With the widespread application of agricultural drones for pesticide spraying, the setting of droplet size and the density of pesticide droplet coverage on crop leaves after spraying have become important concerns for plant protection workers. For important small agricultural pests such as aphids, whiteflies, planthoppers, and leafhoppers, simulating the behavioral responses of these tiny insects after pesticide droplet deposition in the field requires traditional bioassay methods, such as spot methods, leaf dipping methods, and Potter's tower spraying. These methods can only provide a general understanding of the pest population's toxicity response range to pesticides, but cannot truly reveal the specific behavioral differences between the pesticide deposition sites and non-toxic sites on the leaves. This invention combines the pesticide deposition area with the feeding area of the test insects, significantly increasing the probability of the test insects contacting the pesticide deposits. By reducing the activity area of the test insects and expanding the contact range between the test insects and the pesticide deposit area, it helps to improve the success rate of behavioral observation. Compared to traditional bioassay methods, such as the Potter spray method, which makes it difficult to determine the actual utilization rate of insecticide droplets due to the large range of insect activity and the difficulty in observing pesticide spots on leaves, the leaf immersion method, which creates a pesticide-covered area across the entire field, cannot reveal the insects' selective behavior towards pesticide-covered and pesticide-free areas, thus failing to simulate the real conditions in the field. Summary of the Invention
[0003] In view of this, the purpose of this invention is to provide a device and method for observing the behavioral response of micro-insects to insecticide deposits. This device can be combined with a photographic microscope to visually record the behavioral response of micro-insects after contact with insecticide deposits of different morphologies. This will help guide the selection of appropriate droplet size and effective droplet coverage density range when controlling micro-insects in the field, providing valuable reference for agricultural research.
[0004] In addition, the device in this scheme has a simple structure and is easy to operate. It can intuitively demonstrate the selective behavior of tiny piercing-sucking insects when they come into contact with pesticide deposits. It provides a simple device and detection method with reference value for setting appropriate pesticide application conditions.
[0005] To achieve the above-mentioned technical objectives, the technical solution adopted by this invention is as follows: A device for observing the behavioral responses of microscopic insects to insecticide deposits includes an observation box and a feeding tray. Both the observation box and the feeding tray are made of transparent material. The observation box is a box-shaped shell structure with an open lower end and ventilation holes on its upper side wall. The feeding tray is a plate-like structure with a feeding cavity in the middle of its upper end. The upper end of the feeding cavity is open, and attractant feed is placed inside the feeding cavity to form a feeding area. A parafilm is provided on the upper surface of the feeding tray corresponding to the feeding cavity. Insecticide deposits are deposited on the surface of the parafilm outside the feeding cavity. The observation box is placed on top of the feeding tray, covering the feeding cavity, and the upper surface of the feeding tray and the interior of the observation box together form an activity chamber for insects to move around in.
[0006] As a possible implementation, the solution further includes a tray that covers the lower end of the feed pan from bottom to top and encloses the feed cavity therein. The area of the tray opposite the feed cavity is used to place a visual attractant.
[0007] As a preferred implementation method, the observation box of this solution is provided with tongues on both sides of its lower end face, and the feed tray area on both sides of the feed cavity is provided with slots for the tongues to pass through. The observation box is fixedly connected to the feed tray by the tongues cooperating with the slots.
[0008] As a preferred implementation method, the tray in this solution is preferably a rectangular tray, the slot is through the feed tray, and the tongue at the lower end of the observation box passes through the slot and fits tightly against the inner wall of the tray, so that the observation box, feed tray and tray are relatively fixedly connected.
[0009] As a preferred embodiment, the lower end of the feed cavity in this solution is preferably an open structure, and a sealing film, which is a Parafilm sealing film, is provided on the lower surface of the feed tray corresponding to the feed cavity.
[0010] As a preferred implementation method, the visual attractant in this solution is preferably a plant leaf, and the tray is also used to hold water to form a moisturizing zone.
[0011] In addition, the sealing film on the lower surface of the feed tray can be replaced with filter paper or micropores can be provided so that the water in the tray can be used to enhance the moisture retention of the attractant feed. In addition, the tray can prevent the sealing film on the lower side of the feed tray from directly contacting the work surface during operation, thereby avoiding dust adhesion.
[0012] Based on the above, this solution also proposes the application of the aforementioned device in observing the behavioral responses of microscopic insects to pesticide deposits, including: Prepare the attractant feed for the test insects, fill it into the feed cavity of the feed tray, and then seal both ends of the feed cavity with Parafilm sealing film. A droplet carrying an insecticide is applied to the sealing film at the top of the feed chamber and allowed to air dry to form an insecticide deposit area. This insecticide deposit area serves as a contact behavior observation area, which overlaps with the feeding area of the test insect. The test insect's mouthparts can penetrate the sealing film to feed on the artificial feed in the feed chamber. The test insects are starved, then anesthetized with CO2, and then placed on the upper surface of the feed tray so that they are near the feed cavity. The observation box is then covered, so that the test insects are in an activity chamber formed by the upper surface of the feed tray and the inside of the observation box, which allows the insects to move around. After adding a preset amount of water to the tray, place plant leaves as a visual lure. Then, cover the feed tray from bottom to top, so that it covers the feed cavity, and the visual lure is directly opposite the feed cavity. After the test insects awoke, the number of times the test insects touched the pesticide spot and the time they stayed in the pesticide deposit area within the preset time period were observed and counted, so as to statistically analyze the significant differences between different pesticide deposit treatments.
[0013] As a preferred embodiment, the attractant feed of this scheme comprises, by percentage, 10% host plant leaf homogenate, 0.10% ascorbic acid, 5% sucrose and 4% agar, with the remainder being water.
[0014] In preparing the attractant feed for the test insects, the leaves of the host plant are washed, homogenized, ground, and filtered to obtain plant juice. The plant juice and ascorbic acid are then added to a mixture of melted but not solidified sucrose and agar. After thorough stirring, the material is filled into the feed cavity.
[0015] As a preferred embodiment, the method of applying droplets carrying insecticide to the sealing film at the upper end of the feed cavity includes one of the following: (1) Use a capillary tube to drip onto the sealing film to create droplets carrying insecticide; (2) Cover the area of the feed tray with filter paper containing the feed cavity, then cut out clearance holes in the filter paper corresponding to the feed cavity, and then spray with a spray bottle to create droplets carrying insecticide.
[0016] As a preferred implementation method, the test insect in this scheme is the adult tea green leafhopper, and the insecticide is indoxacarb; the host plant leaves and the plant leaves used as visual attractants are both tea leaves.
[0017] Compared with existing technologies, the present invention, employing the above-described technical solution, has the following advantages: The device has a simple structure and is flexible and convenient to operate. This device combines the insecticide deposition spot with the feeding area of the test insect, significantly increasing the probability of the test insect contacting the spot. Furthermore, the miniaturization of the device reduces the activity range of the test insect, improving the success rate of observing the contact behavior of the spot. The device is readily available and easily customized, providing reliable and reproducible test results, making it highly suitable for optimizing application conditions before applying insecticides to control small insects. Based on this device, by observing the behavioral responses of small piercing-sucking insects to insecticide deposition spots, it can provide positive and feasible research assistance for clarifying the influence of different insecticide deposition spot conditions on the behavioral responses of small piercing-sucking insects and the accurate toxicity effect after 48 hours. It also provides valuable reference for selecting appropriate droplet sizes when using drones for insecticide spraying. Attached Figure Description
[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0019] Figure 1 This is a simplified three-dimensional schematic diagram of the implementation structure of the device in this scheme. The transparent parts of the device are hidden, and the dashed lines are schematic diagrams of the positions of the sealing film, feed chamber and attractant feed. Figure 2 This is a simplified three-dimensional exploded view of the implementation structure of the device in this scheme, in which the dashed lines show the transparent material's outline that can be seen through; Figure 3 This is a three-dimensional perspective schematic diagram of the structural components of the device in this scheme, in which the observation box is hidden, as are the transparent parts of the device that can be seen through. Figure 4 This is a simplified three-dimensional exploded view of the implementation structure of the device in this scheme, in which the transparent parts of the device are hidden. Figure 5 This is a diagram illustrating insecticide deposits on the sealing film of this solution; Figure 6 This is a two-dimensional cross-sectional view of the simplified implementation structure of the device in this scheme; Figure 7 This is a schematic diagram of the device in actual use, in which... Figure 7 A is a physical diagram of the device in this scheme. Figure 7 B is one of the photos of the tea green leafhopper moving along the edge of the sealing film. Figure 7C is the second photo of the tea green leafhopper moving along the edge of the sealing film. Detailed Implementation
[0020] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be particularly noted that the following embodiments are for illustrative purposes only and do not limit the scope of the invention. Similarly, the following embodiments are only some, not all, embodiments of the present invention, and all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0021] like Figures 1 to 6 As shown in one embodiment, this embodiment proposes a device for observing the behavioral response of microscopic insects to insecticide deposits. It includes an observation box 1 and a feeding tray 2, both made of transparent material. The observation box 1 is a box-shaped shell structure with an open lower end and ventilation holes 11 on its upper sidewall. The feeding tray 2 is a plate-like structure with a feeding cavity 21 in the middle of its upper end. The upper end of the feeding cavity 21 is open, and attractant feed 3 is placed inside to form a feeding area. A sealing film 31 is provided on the upper surface of the feeding tray 2 corresponding to the feeding cavity 21. The sealing film is Parafilm (which has good tensile strength). Insecticide deposits are deposited on the surface of the sealing film 31 outside the feeding cavity 21 (see reference). Figure 5 or Figure 7 B Figure 7 (As indicated by the arrow in C), the observation box 1 is placed on the feed tray 2, covering the feed cavity 21 therein, and the upper surface of the feed tray 2 and the interior of the observation box 1 together form an activity chamber 13 for insects to move around.
[0022] In addition to the above, the device of this solution also includes a tray 4, which covers the lower end of the feed tray 2 from bottom to top and covers the feed cavity 21 therein. The area of the tray 4 opposite to the feed cavity 2 is used to place the visual attractant 41.
[0023] In this embodiment, the lower end of the feed cavity 21 is an open structure, and the lower surface of the feed tray 2 is provided with a sealing film 32 corresponding to the feed cavity 21. The sealing film 32 is a Parafilm sealing film (or Parafilm, which has good tensile properties). In this way, the visual attractant 41 can be easily discovered by insects. Preferably, the visual attractant 41 in this embodiment is a plant leaf, and the tray 2 is also used to hold water to form a moisturizing area 42 for the plant leaf.
[0024] In addition, the sealing film 32 on the lower surface of the feed tray 2 can be replaced with filter paper or have micropores to allow the water in the tray to enhance the moisture retention of the attractant feed 3. Furthermore, the tray 2 can prevent the sealing film 32 on the lower side of the feed tray 2 from directly contacting the work surface during operation, thereby preventing dust from adhering.
[0025] To facilitate the assembly and connection of the device, preferably, the observation box 1 of this solution is provided with tongues 12 on both sides of the lower end face, and the feed tray 2 area on both sides of the feed chamber 21 is provided with slots 22 for the tongues 12 to pass through. The observation box 1 is fixedly connected to the feed tray 2 by the tongues 12 cooperating with the slots 22.
[0026] The tray 2 described in this solution is a rectangular tray, and the slot 22 passes through the feed tray 2. After the tongue 12 at the lower end of the observation box 1 passes through the slot 22, it is tightly attached to the inner wall of the tray 2, so that the observation box 1, the feed tray 2 and the tray 3 are relatively fixedly connected.
[0027] As an example of one implementation specification of the device in this scheme, the observation box 1 can be 24×24×15mm in length, width and height, and the tongue at its lower end is 8mm higher; the feed tray 2 can be 40×40×5mm in length, width and height, and the feed cavity 21 therein has a diameter of 10mm; the pallet can be 24×24×5mm in length, width and height.
[0028] Based on the above, this solution also proposes the application of the aforementioned device in observing the behavioral responses of microscopic insects to pesticide deposits, which includes the following steps: S01. Prepare the attractant feed for the test insects, fill it into the feed cavity of the feed tray, and let it stand to solidify the attractant feed in the feed cavity (or directly form it on the outside and then fill it in). Then seal both ends of the feed cavity with Parafilm sealing film. The attractant feed consists of the following components by percentage: 10% host plant leaf homogenate, 0.10% ascorbic acid, 5% sucrose and 4% agar, with the remainder being water.
[0029] When preparing the attractant feed for test insects, the leaves of the host plant are washed, homogenized, ground, and filtered to obtain plant juice. The plant juice and ascorbic acid are then added to a mixture of melted but not solidified sucrose and agar. After thorough stirring, the material is filled into the feed cavity.
[0030] S02. Apply droplets of insecticide to the sealing film at the upper end of the feed cavity and allow it to air dry to form an insecticide deposit area. This insecticide deposit area serves as a contact behavior observation area, which overlaps with the feeding area of the test insect. The test insect's mouthparts can penetrate the sealing film to feed on the artificial feed in the feed cavity. The application of insecticide-laden droplets to the sealing film at the upper end of the feed cavity includes one of the following: (1) Use a capillary tube to drop onto the sealing film to create droplets carrying the insecticide; for example, use a capillary tube (0.3 mm × 100 mm) to drop coarse droplets, resulting in a particle size of 570.5 ± 26.5 µm and a coverage density of 7~9 drops / cm. 2 Or use a capillary tube ( Fine droplets were prepared by dotting with 0.1 mm × 100 mm droplets, resulting in a particle size of 250.0 ± 18.5 µm and a coverage density of 55~65 drops / cm². 2 ; (2) Cover the area of the feed tray with filter paper containing the feed cavity, then cut out clearance holes in the filter paper corresponding to the feed cavity, and then spray with a spray bottle to create droplets carrying insecticide. For example: cover the feed well tray with filter paper and cut a circular hole in the feed well area. After 10 mm, use a spray bottle to spray ultrafine droplets onto the feed well area, resulting in a particle size of 80.0 ± 22.5 µm and a coverage density of 75~105 drops / cm². 2 .
[0031] (3) Starve one test worm, then anesthetize it with CO2, and place it on the upper surface of the feed tray so that it is near the feed cavity. Then cover it with the observation box so that the test worm is in the activity room formed by the upper surface of the feed tray and the inside of the observation box. The purpose of only placing one test worm at a time is that in the single-worm isolation observation mode, there are no other test worms to fight with it or other interference behaviors. Multiple test worms in the activity room will greatly reduce the chance of the test worms reaching the contact area of the deposited drug spot, and it is also more likely to cause the isolated test worms to die due to insufficient feeding. (4) After adding a preset amount of water to the tray, place plant leaves as a visual attractant, and then cover the lower end of the feed tray from bottom to top, so that it covers the feed cavity, and the visual attractant is directly opposite the feed cavity. (5) After the test insects are awakened, observe and record them using a microscope or other observation aids (such as camera equipment). Then, count the number of times the test insects touch the pesticide spot and the time they stay in the pesticide spot area within the preset time period, so as to statistically analyze the differences in the significance of different pesticide spot treatments.
[0032] In addition to behavioral observation, this device can also be used to raise test insects for 48 hours, count the survival rate, and compare the significant differences in mortality rates of test insects under different drug-treated conditions over 48 hours.
[0033] As an example of a preferred implementation method, in this scheme, the test insect is an adult tea green leafhopper with a body length of about 3 mm, and the insecticide is indoxacarb; the host plant leaves and the plant leaves used as visual attractants are both tea leaves.
[0034] Combination Figure 7 As shown, this embodiment utilizes the device to create two types of drug spots of varying coarseness using capillary dripping. The drug is 750 mg / L indoxacarb. The number of times the tea green leafhopper adults stayed under the two drug spot conditions did not differ significantly. P = 0.36), but after 48 h, the mortality rate of the large drug spot treatment was significantly higher than that of the small drug spot treatment ( t = 3.02, df = 4, P = 0.04), behavioral observation revealed that the tea green leafhopper would repeatedly move around on the large pesticide residue area when its forelegs and midlegs came into contact with it. Because the large pesticide residue area was relatively large, the actual contact time between the insect and the toxic area was also relatively long. When the pesticide was Beauveria bassiana (concentration 1.32 × 10⁻⁴), 9 When the tea green leafhopper was treated with large and small drug spots (spores / mL), the mortality rate was similar to that of the control. Behavioral observation also showed that the tea green leafhopper had a significant aversion and alarm response to high concentrations of Beauveria bassiana residue drug spots.
[0035] As an example of expanding the method of observing and statistically analyzing test insects, this device can be combined with a camera to observe and record test insects in the observation box. Then, based on the acquired image data, an edge detection algorithm is used to track the test insects. By using the relationship between the position of the drug spot on the sealing film and the position of the test insect, the device can assist in statistically analyzing the number of times the test insect comes into contact with the drug spot and the time spent there. Alternatively, it can be used to mark time points within the image data so that experimenters can directly extract effective data segments to trace the behavior of the test insects. This is a solution currently used in other fields. This embodiment cleverly introduces it into the scenario of test insect observation, and its principle will not be elaborated here.
[0036] The above description is only a part of the embodiments of the present invention and does not limit the scope of protection of the present invention. Any equivalent device or equivalent process transformation made based on the content of the present invention specification and drawings, or direct or indirect application in other related technical fields, are similarly included within the patent protection scope of the present invention.
Claims
1. A device for observing the behavioral responses of microscopic insects to insecticide deposits, characterized in that, It includes an observation box and a feeding tray, both made of transparent material. The observation box is a box-shaped shell structure with an open lower end and ventilation holes on its upper side wall. The feeding tray is a plate-shaped structure with a feeding cavity in the middle of its upper end. The upper end of the feeding cavity is open, and attractant feed is placed inside the feeding cavity to form a feeding area. A sealing film is provided on the upper surface of the feeding tray corresponding to the feeding cavity. Insecticide spots are deposited on the surface of the sealing film outside the feeding cavity. The observation box is placed on the feeding tray, covering the feeding cavity, and the upper surface of the feeding tray and the interior of the observation box together form an activity chamber for insects to move around.
2. The apparatus for observing the behavioral response of microscopic insects to insecticide deposits as described in claim 1, characterized in that, The sealing film is Parafilm sealing film; The device also includes a tray that covers the lower end of the feed tray from bottom to top and covers the feed cavity therein. The area of the tray opposite the feed cavity is used to place a visual attractant.
3. The apparatus for observing the behavioral response of microscopic insects to pesticide deposits as described in claim 2, characterized in that, The observation box has tongues on both sides of its lower end face, and the feed tray areas on both sides of the feed chamber have slots for the tongues to pass through. The observation box is fixedly connected to the feed tray by the tongues engaging with the slots.
4. The apparatus for observing the behavioral response of microscopic insects to pesticide deposits as described in claim 3, characterized in that, The tray is a rectangular tray, the slot is through the feed tray, and the tongue at the bottom of the observation box passes through the slot and fits tightly against the inner wall of the tray, so that the observation box, feed tray and tray are relatively fixedly connected.
5. The apparatus for observing the behavioral response of microscopic insects to pesticide deposits as described in any one of claims 2 to 4, characterized in that, The lower end of the feed chamber is an open structure, and a sealing film, which is Parafilm, is provided on the lower surface of the feed tray corresponding to the feed chamber.
6. The apparatus for observing the behavioral response of microscopic insects to pesticide deposition spots as described in claim 5, characterized in that, The visual attractant is plant leaves, and the tray is also used to hold water to form a moisturizing zone.
7. The application of the device as described in claim 6 in observing the behavioral response of microscopic insects to pesticide deposition spots, characterized in that, It includes: Prepare the attractant feed for the test insects, fill it into the feed cavity of the feed tray, and then seal both ends of the feed cavity with Parafilm sealing film. Apply droplets of insecticide to the sealing film at the top of the feed chamber and allow it to air dry to form an insecticide deposit area. The test insects were starved, then anesthetized with CO2, and then placed on the upper surface of the feed tray, so that they were near the feed cavity. The observation box was then covered, so that the test insects were in the activity chamber formed by the upper surface of the feed tray and the inside of the observation box. After adding a preset amount of water to the tray, place plant leaves as a visual lure. Then, cover the feed tray from bottom to top, so that it covers the feed cavity, and the visual lure is directly opposite the feed cavity. After the test insects awoke, the number of times the test insects touched the pesticide spot and the time they stayed in the pesticide deposit area within the preset time period were observed and counted, so as to statistically analyze the significant differences between different pesticide deposit treatments.
8. The application as described in claim 7, characterized in that, The attractant feed comprises, by percentage, 10% host plant leaf homogenate, 0.10% ascorbic acid, 5% sucrose and 4% agar, with the remainder being water; When preparing the attractant feed for test insects, take the leaves of the host plant, wash them, homogenize, grind and filter them to obtain plant juice. Then add the plant juice and ascorbic acid to the melted but not solidified mixture of sucrose and agar. After stirring thoroughly, fill the feed cavity with the material.
9. The application as described in claim 7, characterized in that, Applying droplets carrying insecticide to the sealing film at the upper end of the feed cavity includes one of the following: (1) Use a capillary tube to drip onto the sealing film to create droplets carrying insecticide; (2) Cover the area of the feed tray with filter paper containing the feed cavity, then cut out clearance holes in the filter paper corresponding to the feed cavity, and then spray with a spray bottle to create droplets carrying insecticide.
10. The application as described in claim 7, characterized in that, The test insect was an adult tea green leafhopper, and the insecticide was indoxacarb; both the host plant leaves and the plant leaves used as visual attractants were tea leaves.