Device and method for measuring bee smell preference behavior

Abstract

The invention discloses a device and a method for measuring bee smell preference behaviors, and relates to the technical field of bee smell localization, the device comprises a bottom plate, a cover plate, a smell source bottle and a camera device, the cover plate is arranged above the bottom plate, a retention cavity is arranged between the cover plate and the bottom plate, and the camera device is arranged in the retention cavity. A retention cavity is formed in the bottom plate, a plurality of channels distributed in a radial shape with the retention cavity as the center are further arranged between the cover plate and the bottom plate, the ends, facing the retention cavity, of the channels communicate with the retention cavity, the ends, away from the retention cavity, of the channels are provided with the flavor source bottles, and the cover plate is provided with an inlet and outlet communicating with the retention cavity; the upper end of the channel is transparent, the camera device is arranged above the cover plate, and a camera of the camera device faces the cover plate. According to the method, the preference of bees to different compounds can be measured, and behavior selection of bees to the compound flavor sources can be objectively and accurately evaluated.

Description

Technical Field

[0001] The present invention relates to the technical field of honeybee olfactory localization, and in particular to a device and method for measuring honeybee odor preference behavior. Background Art

[0002] As pollinators of numerous plants, bees play a vital role in maintaining ecological balance. They are the world's primary agricultural pollinator, pollinating over 85% of crops. Bees have evolved over 150 million years, their emergence closely tied to the flourishing of flowering plants in the Late Cretaceous. Over millions of years of evolution, bees and plants have developed a mutually beneficial symbiotic relationship. Flowering plants and bees enjoy a uniquely mutually beneficial relationship: bees obtain food, which in turn facilitates pollination and improves fruit yield and quality.

[0003] As the most important pollinating and economic insects, bees play an important role in crop production and the global ecological environment. The volatile odors of flowers are usually species-specific. Bees distinguish plants by identifying one or several key compounds in a complex mixture of odors. These active substances can directly affect the bees' flower-visiting behavior. At present, it is still unclear which plants release specific odors that are associated with food during the long-term co-evolution of bees and flowering plants. In addition, how do bees determine the optimal state and food content of plant flowers for collection before collecting? And during pollination, what specific food signals do plants emit to attract bees for pollination? These scientific problems need to be solved urgently. Therefore, it is of great significance and prospects to screen out the key chemical components for bees to communicate information with the flowers of pollinating plants and to deeply explore the interaction between bees and pollinating plants. Summary of the Invention

[0004] The purpose of the present invention is to provide a device and method for measuring the odor preference behavior of bees to solve the problems existing in the above-mentioned prior art. It can measure the preference of bees for different compounds and objectively and accurately evaluate the behavioral selection of bees for compound odor sources.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides a device for measuring the odor preference behavior of bees, comprising a base plate, a cover plate, a flavor source bottle, and a camera device. The cover plate is arranged above the base plate, a retention cavity is provided between the cover plate and the base plate, and a plurality of channels are further provided between the cover plate and the base plate, the channels being radially distributed with the retention cavity as the center. The ends of the channels facing the retention cavity are connected to the retention cavity, the flavor source bottle is provided at the ends of the channels away from the retention cavity, an entrance and exit communicating with the retention cavity are provided on the cover plate, the upper ends of the channels are transparent, and the camera device is provided above the cover plate with its camera facing the cover plate.

[0007] In one embodiment, the distance between the bottom plate and the cover plate is 20-30 mm.

[0008] In one embodiment, six channels are provided, and the six channels are distributed at equal angles in a circumferential direction with the retention chamber as the center.

[0009] In one embodiment, the inlet and outlet are circular holes with a diameter of 30-50 mm.

[0010] In one embodiment, the channel is formed by two parallel rectangular long plates, the bottom plate and the cover plate, and the two rectangular long plates are fixed on the bottom plate.

[0011] In one embodiment, the cover plate support is placed on the upper end of each of the rectangular long plates.

[0012] In one embodiment, the bottom plate, the cover plate and the rectangular strip plate are made of transparent acrylic plates.

[0013] In one embodiment, the inlet and outlet are provided with a sealing plug.

[0014] In one embodiment, the bottom plate and the cover plate are circular plates with equal diameters.

[0015] The present invention also provides a method for measuring the odor preference behavior of honey bees, based on the above-mentioned device for measuring the odor preference behavior of honey bees, comprising the following steps:

[0016] (1) starving the bees;

[0017] (2) placing a bee into the retention chamber through the entrance and exit, closing the entrance and exit, and starting recording with the camera device;

[0018] (3) Record the choices made by the bees on the compounds in the odor source bottles in each channel, and calculate the number and frequency of choices for each compound.

[0019] Compared with the prior art, the present invention has achieved the following technical effects:

[0020] The present invention provides an apparatus and method for measuring the odor preference behavior of honey bees. A retention chamber is provided between a cover plate and a bottom plate, and multiple channels are radially distributed with the retention chamber as the center. The end of each channel facing the retention chamber is connected to the retention chamber. An odor source bottle is provided at the end of each channel away from the retention chamber. Different compounds are placed in the odor source bottle. Honey bees are placed into the retention chamber through an entrance and exit on the cover plate. The bees' choices of compounds in each channel are recorded by a camera device, and the number and frequency of choices for each compound are calculated, thereby screening out odor compounds with specific preferences and objectively and accurately evaluating the honey bees' behavioral choices for compound odor sources. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0022] Figure 1 Schematic diagram of the connection structure between the bottom plate and the cover plate in an embodiment of the present invention;

[0023] Figure 2 for Figure 1 A top view of

[0024] Figure 3 Schematic diagram of the structure of the bottom plate in an embodiment of the present invention;

[0025] Figure 4 for Figure 1 sectional view of

[0026] Figure 5 This is a statistical chart of the number of visits of bees to different odor compounds in the experiment of the embodiment of the present invention.

[0027] In the figure: 1-bottom plate, 2-cover plate, 3-retention chamber, 4-channel, 5-inlet and outlet, 6-rectangular long plate. DETAILED DESCRIPTION

[0028] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0029] The purpose of the present invention is to provide a device and method for measuring the odor preference behavior of honeybees to solve the problems existing in the prior art. It can measure the preference of honeybees for different compounds and objectively and accurately evaluate the behavioral selection of honeybees for compound odor sources.

[0030] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0031] like Figures 1-4 As shown, the present invention provides a device for measuring the odor preference behavior of bees, including a base plate 1, a cover plate 2, a flavor source bottle and a camera device, the cover plate 2 is arranged above the base plate 1, a retention chamber 3 is provided between the cover plate 2 and the base plate 1, and a plurality of channels 4 are further provided between the cover plate 2 and the base plate 1, which are radially distributed with the retention chamber 3 as the center, and each channel 4 is connected to the retention chamber 3 at one end facing the retention chamber 3, and a flavor source bottle is provided at one end of each channel 4 away from the retention chamber 3, an entrance and exit 5 connected to the retention chamber 3 is provided on the cover plate 2, the upper end of the channel 4 is transparent, and the camera device is arranged above the cover plate 2 with its camera facing the cover plate 2, and the focal length of the camera device covers the entire cover plate 2.

[0032] During use, bees are placed into the retention chamber 3 through the entrance 5 on the cover 2. A camera records the bees' choices of compounds in each channel and calculates the number and frequency of selection for each compound. This allows for the selection of odor compounds with specific preferences and an objective and accurate assessment of the bees' behavioral preferences for compound odor sources. This device is inexpensive, portable, and can be used to rapidly conduct a large number of experiments.

[0033] The distance between the bottom plate 1 and the cover plate 2 is 20-30 mm, preferably 20 mm, so that the bees can move smoothly in the channel 4.

[0034] There are six channels 4, which are distributed at equal angles in the circumferential direction with the retention chamber 3 as the center. In actual use, other different numbers of channels 4 can also be set according to use requirements.

[0035] The entrance and exit 5 is a circular hole with a diameter of 30-50 mm, preferably 30 mm, which is convenient for placing bees into the retention chamber 3.

[0036] Channel 4 is formed by two parallel rectangular strips 6, which are enclosed by the base plate 1 and the cover plate 2. The two rectangular strips 6 are fixed to the base plate 1. The rectangular strips 6 are glued to the base plate 1. The rectangular strips 6 are 200 mm long, 20 mm wide, and 3 mm thick. The flavor source bottle can also be glued to the outer end of channel 4.

[0037] The cover plate 2 is supported and placed on the upper end of each rectangular long plate 6, and the cover plate 2 is supported by the rectangular long plate 6. The cover plate 2 can be removed to facilitate cleaning the internal structure and also to facilitate placing the odor source sample into the device.

[0038] The bottom plate 1, the cover plate 2 and the rectangular strip plate 6 are made of transparent acrylic plates.

[0039] The entrance and exit 5 is provided with a sealing plug to prevent the bees from escaping from the retention chamber 3 .

[0040] The bottom plate 1 and the cover plate 2 are circular plates with the same diameter, 490 mm in diameter and 3 mm in thickness.

[0041] Based on the above-mentioned device for measuring the odor preference behavior of honey bees, the present invention also provides a method for measuring the odor preference behavior of honey bees, comprising the following steps:

[0042] (1) The honey bees required for the experiment were healthy and fully developed worker bees. The worker bees were captured from the vicinity of the beehive and starved for 24 hours before the experiment.

[0043] (2) Place 10 healthy bees into the retention chamber 3 through the entrance 5 of the cover plate 2, close the entrance 5, and start recording with a camera;

[0044] (3) The experiment lasted 20 minutes. The bees' choices of the compounds in the odor source bottles in each channel 4 were recorded, and the number and frequency of choices for each compound were calculated.

[0045] The present invention will be further explained below with reference to specific embodiments.

[0046] Example 1:

[0047] The volatile compounds in Tilia amurensis flowers were identified using dynamic headspace adsorption and gas chromatography-mass spectrometry. The adsorption method involved connecting a gas sampling tube filled with 150 mg of Tenax-TA adsorbent to an air sampler (20-01-BD; BMILP, Beijing). The released VOCs were adsorbed into the sampling tube via an airflow. The air sampler's flow meter was adjusted to a flow rate of 500 mL / min, and adsorption was continued for 2 hours. A blank sampling bottle was also collected as a blank control. The gas chromatography-mass spectrometry procedure used was as follows: HP-5MS column (0.25 mm × 30 mm × 0.25 μm); inlet temperature: 240°C; helium carrier gas: purity ≥99.99%; flow rate: 1.0 mL / min. The column oven program started at 45°C, held for 5 minutes, then increased from 45°C to 130°C at 6°C / min, then from 130°C to 240°C at 10°C / min, and finally held at 240°C for 8 minutes. The injection method was splitless. Mass spectrometry parameters were: ion source temperature: 230°C; interface temperature: 250°C. Ionization mode was electron ionization (EI). A full scan was performed over the mass range of m / z 45 to 500. Volatile compounds were searched and identified using the NIST14 mass spectral library, and the relative content of each component was analyzed by area normalization.

[0048] The olfactory active substances were determined by electroantennogram technique, and the method was as follows:

[0049] Sample preparation for the volatile components identified by GC-MS (gas chromatography-mass spectrometry) involved diluting purchased standard compounds in liquid paraffin to produce six different dilutions (0 μg / μL, 10 μg / μL, 100 μg / μL, 200 μg / μL, 300 μg / μL, and 400 μg / μL). Bees were fasted for 24 hours before electroantenniograph (EAG) measurements. The bees were placed under a microscope, and the antennae were removed from the base using a planer. A reference glass electrode filled with Ringer's solution was placed at the base and contacted with an inserted Ag / AgCl wire. Next, a small incision was made at the top of the antennae using iris scissors to expose the internal tissue. A recording glass electrode, also filled with Ringer's solution, was inserted into the incision.

[0050] EAG responses were detected using a combination probe. DC potentials were recorded (universal AC / DC probes) and processed and analyzed using EAG 2000 software (Syntech, Netherlands). The test odor samples were provided by an air stimulus controller (CS55; Syntech, Netherlands) with a constant airflow of 300 mL / min. The test antenna was adjusted to the center of the open end of the metal tube so that the air carried by the compound was uniform. The concentration of each compound was tested at intervals of 1 min for a duration of 0.5 s. The time interval between each sample was 30 s. All compounds were tested from low to high concentrations to avoid olfactory adaptation. Standard liquid paraffin stimulation was performed at the beginning and end of each recording as a blank control for antennal responses. Twelve replicates were performed for each sample, and the antennae were replaced after each sample was measured. They were cut immediately before use to ensure that the antennae were fresh.

[0051] The results of the antennal potential experiment are expressed as the absolute value of EAG, and the formula is as follows:

[0052]

[0053] Based on the EAG test results, the method for studying the volatile compound preferences of bees during their visit to Tilia amurensis flowers is as follows: the key active compounds with the strongest EAG responses were selected: nonanal, hexanal, heptanal, methylheptenone and 3-methyl-2-butenal. Before starting the experiment, the maze bottom plate (i.e., bottom plate 1) was heated to about 30°C using a heating pad to prevent bees from gathering and keeping warm during the experiment. A compound was placed in each channel 4 of the maze system (a system of radially distributed channels 4 formed by the cover plate 2 and the bottom plate 1) as an odor source. In each experiment, ten bees were captured from the beehive and placed through the circular hole on the cover plate 2 of the maze system. The camera was turned on to record the number of visits of the bees to different compounds. A valid visit was when the bee approached the odor source bottle, touched the top of the odor source bottle with its head, and left. All other cases were invalid visits, that is, they were not counted in the total cumulative number. Each experiment lasted twenty minutes and was repeated four times. The maze system was cleaned after each repetition, and the order of placing the odor source bottles was different each time. See Figure 5 , which is a statistical graph of the number of visits of bees to different odor compounds in the experiment.

[0054] The device and method for measuring honeybee odor preference provided by this invention are used to rapidly screen honeybees' preference for different volatile odorants. By leveraging the differences in the responses of honeybee antennae to different odors, the device can identify odor compounds with specific preferences. The device is inexpensive, portable, and can quickly conduct large-scale experiments.

[0055] The present invention uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only intended to help understand the method and core concept of the present invention. At the same time, those skilled in the art will find that the specific implementation methods and application scopes may vary based on the concept of the present invention. In summary, the contents of this specification should not be construed as limiting the present invention.

Claims

1. A device for measuring honeybee odor preference behavior, characterized by: The invention comprises a bottom plate, a cover plate, a flavor source bottle and a camera device, wherein the cover plate is arranged above the bottom plate, a retention cavity is provided between the cover plate and the bottom plate, and a plurality of channels are further provided between the cover plate and the bottom plate and are radially distributed with the retention cavity as the center, and each of the channels is connected to the retention cavity at one end facing the retention cavity, and the flavor source bottle is provided at one end of each channel away from the retention cavity, an entrance and exit connected to the retention cavity is provided on the cover plate, the upper end of the channel is transparent, and the camera device is provided above the cover plate with its camera facing the cover plate.

2. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: The distance between the bottom plate and the cover plate is 20-30 mm.

3. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: There are six channels, and the six channels are distributed at equal angles in the circumferential direction with the retention chamber as the center.

4. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: The inlet and outlet are circular holes with a diameter of 30-50 mm.

5. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: The channel is formed by two parallel rectangular long plates, the bottom plate and the cover plate, and the two rectangular long plates are fixed on the bottom plate.

6. The device for measuring the odor preference behavior of honey bees according to claim 5, characterized in that: The cover plate support is placed on the upper end of each of the rectangular long strips.

7. The device for measuring honeybee odor preference behavior according to claim 5, characterized in that: The bottom plate, the cover plate and the rectangular long strip plate are made of transparent acrylic plates.

8. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: The inlet and outlet are provided with sealing plugs.

9. The device for measuring honeybee odor preference behavior according to claim 1, characterized in that: The bottom plate and the cover plate are circular plates with equal diameters.

10. A method for measuring honey bee odor preference behavior, characterized in that: The device for measuring the odor preference behavior of bees according to any one of claims 1 to 9 comprises the following steps: (1) starving the bees; (2) placing a bee into the retention chamber through the entrance and exit, closing the entrance and exit, and starting recording with the camera device; (3) Record the choices made by the bees on the compounds in the odor source bottles in each channel, and calculate the number and frequency of choices for each compound.

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