Three-cage olfactometer and mosquito olfaction selection preference experiment method

Abstract

The invention relates to a three-cage olfactometer and a mosquito olfaction selection preference experiment method, the three-cage olfactometer comprises a first side cage, a middle cage and a second side cage which are arranged in sequence, one side of the middle cage is communicated with the first side cage through a pipeline, and the other side of the middle cage is communicated with the second side cage through a pipeline; the first side cage is communicated with a first test mouse container through a pipeline; the first side cage is communicated with a first test mouse container through a pipeline, the second side cage is communicated with a second test mouse container through a pipeline, and the first test mouse container and the second test mouse container are respectively communicated with an air supply device through a pipeline; an exhaust port is formed in the top of the middle cage. The experiment device is high in flexibility and can be disassembled, assembled, transformed and the like according to experiment requirements. In the experiment process, the experiment result can be visually seen, the reliability of the mosquito counting result is high, the experiment is easy to operate and repeatable, and the result accuracy is high.

Description

Technical Field

[0001] The invention belongs to the technical field of experimental equipment, and in particular relates to a three-cage olfactometer and a mosquito olfactory selection preference experimental method. Background Art

[0002] Aedes mosquitoes are an important disease vector in nature, capable of transmitting a variety of diseases such as dengue fever, Zika virus disease and chikungunya fever, bringing a huge burden to global public health. In recent years, the dengue fever epidemic in my country has mainly been caused by local transmission of cases imported from abroad, with the main source countries being Southeast Asian and South Asian countries such as Myanmar, Cambodia, and Indonesia; most of them are distributed in coastal or border areas such as Guangdong and Yunnan. Affected by globalization, urbanization and climate change, the prevalence of mosquito-borne viral infectious diseases continues to expand around the world, and the number of infected people is also increasing year by year, and the prevention and control situation remains severe.

[0003] Mosquitoes find hosts through the smell of human skin and exhaled CO2, and accurately locate the location of blood sucking based on the composition of human sweat. Research by Zhao et al. shows that the brain of Aedes aegypti has more than 60 nerve centers called globules, but only two globules respond to human odor mixtures (mainly mixtures characterized by decanal), and the signals emitted by these two globules sensitive to human odor can also significantly enhance the behavior of mosquitoes looking for humans. This characteristic of Aedes aegypti also makes it an important vector of insect-borne viruses. Mosquitoes mainly receive chemical molecular signals through receptors encoded by three multigene families, namely odorant receptors (Odorant receptor, OR), ionotropic receptors (Ionotropic receptor, IR) and gustatory receptors (Gustatory receptor, GR). OR is an odor-gated ion channel, formed by a heteropolymer complex of a conserved odor co-receptor (Orco) and a ligand-selective OR, and mainly responds to esters, alcohols, ketones and aldehydes. IR is an ionotropic glutamate receptor composed of one or more of three conserved co-receptors Ir25a, Ir8a and Ir76b, which mainly responds to carboxylic acids and amines. Although GR is a taste receptor of mosquitoes, studies have shown that Gr3 of Aedes aegypti is the main receptor for recognizing CO2, and Aedes aegypti with mutations in the Gr3 gene show insensitivity to CO2.

[0004] Under the action of these receptors, mosquitoes can not only accurately locate the location of humans, but also find the type of people they "prefer" in the vast sea of ​​people. De Obaldia et al. found that the different attractiveness of humans to mosquitoes is closely related to the level of carboxylic acid produced by human skin. People who are highly attractive to mosquitoes have higher expression of carboxylic acid on their skin. And the odor molecules produced by humans that are highly attractive to mosquitoes are far more than just carboxylic acid. Researchers Cheng Gong's team has shown that after mice are infected with dengue virus or Zika virus, the skin symbiotic bacteria Bacillus genus expands and produces more acetophenone, which makes them more attractive to mosquitoes. In addition, Ghaninia et al.'s research shows that the difference in human attractiveness to mosquitoes may be related to CO2 and acetone and a mixture of the two, and acetone alone can still have a strong attraction to Aedes aegypti. In addition, the content of lactic acid in human skin residues is 10-100 times that of other animals. Studies have shown that the use of enzymes to remove lactic acid from the skin surface greatly reduces the attractiveness of humans to female Aedes aegypti mosquitoes, while adding lactic acid to animal odor extracts will enhance their attractiveness to female mosquitoes. A variety of volatile gases produced during human metabolic activities are attractive to mosquitoes, and different people have different attractiveness to mosquitoes due to different metabolism. However, which odor molecules have an attractive effect and the mechanism of action of these odor molecules on mosquitoes still need further exploration. Summary of the invention

[0005] The invention aims to provide a three-cage olfactometer and a mosquito olfactory selection preference experimental method.

[0006] To achieve the above-mentioned purpose and other related purposes, the technical solution provided by the present invention is: a three-cage olfactometer, comprising a first side cage, a middle cage and a second side cage arranged in sequence, one side of the middle cage is connected to the first side cage through a pipeline, and the other side of the middle cage is connected to the second side cage through a pipeline; the first side cage is connected to a first test mouse container through a pipeline; the second side cage is connected to a second test mouse container through a pipeline, and the first test mouse container and the second test mouse container are respectively connected to an air supply device through a pipeline; an exhaust port is provided on the top of the middle cage.

[0007] The preferred technical solution is: an air flow meter is provided on the pipeline connecting the first side cage and the first test mouse container; an air flow meter is provided on the pipeline connecting the second side cage and the second test mouse container; and the air supply device is an air pump.

[0008] The preferred technical solution is: an air filter is provided on the pipeline connecting the first side cage and the first test mouse container; an air filter is provided on the pipeline connecting the second side cage and the second test mouse container.

[0009] The preferred technical solution is: the air filter is filled with activated carbon.

[0010] The preferred technical solution is: the first test mouse container and the second test mouse container are a conical flask, and the conical flask is provided with an air inlet and an exhaust port.

[0011] To achieve the above purpose and other related purposes, the technical solution provided by the present invention is: an experimental method for conducting olfactory selection preference of mosquitoes, the experimental device adopts the above three-cage olfactometer, and the experimental method comprises the following steps: Step 1: Connect all parts and ventilate to check whether the equipment is completely airtight; Step 2: gas is supplied into the pipeline by the gas supply device, and the gas enters the first test mouse container and the second test mouse container respectively; the gas passing through the first test mouse container and the second test mouse container respectively enters the first side cage and the second side cage through the gas flow meter; the gas is discharged through the exhaust port at the top of the middle cage; Step 3: Before each experiment begins, ventilate all parts after they are connected to eliminate odors.

[0012] Step 4: Two different kinds of test mice are placed into the first test mouse container and the second test mouse container through the upper ports of the first test mouse container and the second test mouse container, respectively, and then the test mosquitoes are placed into the middle cage of the three-cage olfactometer; start timing; Step 5: Record the number of mosquitoes flying against the wind to the first side cage and the second side cage at different times after the start of the experiment; Step 6: After the experiment, ventilate to remove odor.

[0013] Due to the application of the above technical solution, the present invention has the following advantages compared with the prior art: 1. The present invention is a detachable three-cage olfactometer, which is mainly used for mosquito olfactory selection preference experiments. The cubic sub-chocolate transparent box, cylindrical barrel wall, airflow meter, conical bottle, Y-tube, activated carbon and air pump that constitute the whole set of equipment can be disassembled independently of each other. The three-cage olfactometer device is easy to disassemble and assemble, occupies a small space, and is very convenient for experimental use; in addition, the device has good airtightness and can eliminate the influence of the external environment on the experiment. The experimental device is highly flexible and can be disassembled, assembled and modified according to experimental requirements. During the experiment, the experimental results can be seen intuitively, the mosquito counting results are highly reliable, the experiment is easy to operate and repeatable, and the results are highly accurate.

[0014] 2. In order to explore the selectivity of mosquitoes for different volatile gases of human metabolism, the present invention designed a closed device three-cage olfactometer and conducted a mosquito selection preference experiment. The hair on the back of the mouse was removed, and the same volume of different volatile gas solutions were applied. After the equipment was assembled, the mice applied with different volatile gases were placed in the conical bottles on both sides of the three-cage olfactometer for ventilation. The airflow meter was used to ensure that the airflow into the cages on both sides of the three-cage olfactometer was consistent. The mosquitoes were placed in the middle cage of the three-cage olfactometer and flew against the wind to the left and right cages to make a choice. The number of mosquitoes that made the choice was counted, and the preference of the mosquitoes was recorded. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 Mosquito preference test for diabetic mice.

[0016] Figure 2 Test the mosquito's preference for diabetic mice.

[0017] Figure 3 Mosquitoes' preference for type 2 diabetic mice and normal mice after removing volatile gases. DETAILED DESCRIPTION

[0018] The following is a description of the implementation of the present invention by means of specific embodiments. Those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification.

[0019] See also Figure 1-3 . It should be noted that the structures, proportions, sizes, etc. illustrated in the drawings in this specification are only used to match the contents disclosed in the specification for people familiar with this technology to understand and read, and are not used to limit the limiting conditions for the implementation of the present invention, so they have no substantial technical significance. Any modification of the structure, change in the proportional relationship, or adjustment of the size should still fall within the scope of the technical content disclosed by the present invention without affecting the effects and purposes that can be achieved by the present invention. At the same time, the terms such as "upper", "lower", "left", "right", "middle" and "one" quoted in this specification are only for the convenience of description, and are not used to limit the scope of the implementation of the present invention. Changes or adjustments in their relative relationships should also be regarded as the scope of the implementation of the present invention without substantially changing the technical content.

[0020] Example 1: Three-cage olfactometer and mosquito olfactory preference experimental method (1) The Zhanjiang strain of Aedes aegypti was obtained from the Guangdong Provincial Center for Disease Control and Prevention. The experimental conditions were 28 ± 1 °C, relative humidity 80 ± 5%, 14 h of light, and 10 h of darkness. Larvae were reared at a density of 150–200 / L tap water (standing overnight) and fed with turtle food. Mosquito larvae were fed with turtle food (Yingjin Fish Food Co., Ltd., Shenzhen, China). Adult mosquitoes were raised in 20 cm × 20 cm × 35 cm nylon mesh cages and fed with 10% glucose solution. Blood from Kunming mice provided by the Animal Experiment Center of Anhui Medical University was used to feed adult female mosquitoes to induce egg laying. The use of animals was approved by the Experimental Animal Ethics Committee of Anhui Medical University (approval code: LLSC20210773).

[0021] (2) 6-week-old type 2 diabetes mouse model (BSK-Lepr em2Cd479 / Gpt mice) and control mice (C57 mice) were purchased from Jicui Yaokang Biotechnology Co., Ltd. for experiments. The fasting blood glucose of type 2 diabetic mice was ≥11.1 mmol / L, and the random blood glucose was ≥16.7 mmol / L. The normal blood glucose of the control mice was 4.44 mmol / L-11.1 mmol / L. Before each experiment, the blood glucose of type 2 diabetic mice and normal mice was measured.

[0022] In order to detect whether mosquitoes have a selective preference for type 2 diabetic mice, and to explore whether type 2 diabetic mice are more attracted to mosquito bites and are therefore more susceptible to mosquito-borne infectious diseases.

[0023] The technical route provided by the present invention is: (1) The experiment was carried out using the designed three-cage olfactometer. Type 2 diabetic mice and normal mice were placed in the left and right cages of the three-cage olfactometer respectively. Female Aedes aegypti mosquitoes were placed in the middle cage, gas was introduced, and the number of female mosquitoes flying against the wind to the left and right cages was counted.

[0024] (2) Place type 2 diabetic mice and normal mice in conical flasks on both sides respectively, put Aedes aegypti mosquitoes in the middle cage, introduce gas, and count the number of female mosquitoes flying against the wind to the cages on the left and right sides.

[0025] (3) Add activated carbon between the two conical bottles and the cages on the left and right sides to filter the smell of mice in the conical bottles. Count the number of mosquitoes flying against the wind to the cages on the left and right sides.

[0026] The present invention is the first to use a closed three-cage olfactometer to carry out a mosquito selection preference experiment.

[0027] 1. Before the experiment begins, connect the three-cage olfactometer and ventilate it for 10 minutes to eliminate odor interference. Use mousetraps to fix type 2 diabetic mice and normal mice respectively. Put the two fixed mice into the left and right cages of the three-cage olfactometer through the insect entrances of the left and right cages. Ensure that the device is airtight, and use an airflow meter to keep the airflow on the left and right sides consistent, with an airflow rate of 1L / min. Use a mosquito trap to transfer 100 female mosquitoes from the mosquito cage to the middle cage of the three-cage olfactometer. The experiment begins and the time is counted. The mosquitoes fly against the wind to the left and right cages, and enter the cylindrical barrel walls on both sides to make a choice. Count the number of mosquitoes on both sides 5min, 10min, 20min, and 30min after the start of the experiment.

[0028] The results showed that the number of mosquitoes on the side of the cage containing type 2 diabetic mice was always greater than that on the other side. At 30 minutes, the percentage of mosquitoes on the side containing type 2 diabetic mice and normal mice was 70.8% VS 29.2% ( Figure 1 B).

[0029] (A) Three-cage olfactometer. 100 female Aedes aegypti mosquitoes were placed in the middle cage, and experimental mice were placed in the left and right cages. Female mosquitoes made a choice when they flew against the wind to the left and right cages. (B) Three-cage olfactometer. The left and right cages contained a type 2 diabetic mouse and a normal mouse respectively. The percentage of mosquitoes selected by female mosquitoes against the wind. ** indicates P <0.001.

[0030] 2. Place a type 2 diabetic mouse and a normal mouse in a conical flask respectively. Ensure that the device is sealed, and let air filtered by activated carbon pass through the trachea on both sides. Use an airflow meter to keep the airflow on both sides consistent, with an airflow rate of 1L / min. Use a mosquito trap to transfer 100 female Aedes aegypti mosquitoes to the middle cage of the three-cage olfactometer, and ensure that the device is sealed. Start timing, and the mosquitoes will fly against the wind to the left and right cages, and enter the left and right walls of the olfactometer to make a choice. Count the number of mosquitoes on both sides 5min, 10min, 20min, and 30min after the start of the experiment.

[0031] The results showed that the number of mosquitoes in the cage on one side of the conical flask containing type 2 diabetic mice was always greater than that on the other side. At 30 minutes, the percentage of mosquitoes on the side of type 2 diabetic mice and the side of normal mice was: 96.2% VS 3.8% ( Figure 2 When we placed two mice in the conical flasks on both sides, we obtained similar results. At 30 minutes, the percentage of mosquito numbers on the side of type 2 diabetic mice and the side of normal mice was 89.4% VS 10.6% ( Figure 2 The results showed that mice with type 2 diabetes were more attractive to mosquitoes than normal mice ( P <0.05).

[0032] (A) Three-cage olfactometer. 100 female Aedes aegypti mosquitoes were placed in the middle cage, and experimental mice were placed in the conical bottles on the left and right sides. The female mosquitoes made a choice when they flew against the wind to the cages on the left and right sides. (B) A ventilated conical bottle contained a type 2 diabetic mouse and a normal mouse, and the percentage of mosquitoes selected by the female mosquitoes against the wind. (C) Two type 2 diabetic mice and two normal mice were placed in ventilated glass bottles, and the percentage of mosquitoes selected by the female mosquitoes against the wind. ** indicates P <0.001.

[0033] 3. To further verify whether the volatile gases produced by diabetic mice were responsible, we added filters containing activated carbon and gas chromatography adsorbent filters to the ventilation bottles on both sides of the three-cage olfactometer and the middle of the two cages to adsorb and remove the volatile gases from type 2 diabetic mice and normal mice. Then we transferred the female Aedes aegypti mosquitoes to the middle cage of the three-cage olfactometer and compared their selection results. The results showed that after removing the volatile gases on both sides, there was no statistical difference in the proportion of female mosquitoes choosing type 2 diabetic mice and normal mice ( P >0.05). Combined with the results of the first study, we can conclude that the difference in volatile gases produced by type 2 diabetic mice makes them more attractive to mosquitoes.

[0034] (A) Three-cage olfactometer. Filters containing activated carbon and gas chromatography adsorbent filter cartridges were added between the ventilation bottles on both sides and the cages on both sides to adsorb and remove volatile gases from type 2 diabetic mice and normal mice. 100 mated female mosquitoes were placed in the middle cage for upwind selection. (B) A type 2 diabetic mouse and a normal mouse were placed in the ventilation glass bottles respectively, and the percentage of mosquitoes selected by female mosquitoes upwind. ns means P >0.05.

[0035] Example 2: Three-cage olfactometer and mosquito olfactory preference experimental method The mosquito olfactory preference device is a closed three-cage olfactometer. Mice coated with different volatile gases are placed in conical bottles on both sides, and airflow is introduced. The airflow meter is used to ensure that the airflow entering the left and right cages is the same. Mosquitoes are placed in the middle cage of the three-cage olfactometer. When the mosquitoes fly against the wind to the left and right cages, they make a choice.

[0036] The three-cage olfactometer includes three cube acrylic transparent boxes, two cylindrical barrel walls, two conical bottles, a trachea, a split-flow Y-tube, an airflow meter and an air pump. The cube acrylic transparent boxes and the cylindrical barrel walls are detachable. The equipment can be connected with a vent pipe. The cube acrylic transparent boxes all have insect inlets. The conical bottles can be opened.

[0037] The size of the cube acrylic transparent box is 30×30×30cm, and there is a circular opening with a diameter of 10cm on the front of the box as the insect entrance.

[0038] There is a circular insect inlet on the front, and the insect inlet cover is fixed by screws. There is a rubber ring at the opening to ensure the airtightness of the device.

[0039] There is a circular exhaust port with a diameter of 10 cm on the top of the middle cube acrylic transparent box. The gas enters from the left and right sides of the box and is discharged through the exhaust port on the top.

[0040] Three cube acrylic transparent boxes are connected by two cylindrical barrel walls. There are rubber rings at the joints to ensure the tightness of the device. The joints are fixed by screws and can be tightened manually.

[0041] Cylindrical barrel wall dimensions: bottom diameter 10cm, height 20cm.

[0042] The main body of the device is formed by connecting three cube acrylic transparent boxes with two cylindrical barrel walls. Mosquitoes enter from the insect inlet of the middle cube acrylic transparent box and make a choice when they enter the circular connecting pipes on both sides.

[0043] The airflow meter is connected to the cages on the left and right sides through air pipes of the same length, material and caliber. The airflow meter is a glass rotor flowmeter provided by the sample cotton flow meter factory. The airflow meter has an air inlet below and an air outlet above. After ventilation, the size of the airflow inflow is controlled by controlling the height of the glass rotor.

[0044] The airflow meter is connected to the two conical flasks through an air tube with the same length, material and caliber.

[0045] The upper end of the conical flask can be opened, and the cover contains a rubber ring to ensure the airtightness of the device. The conical flask has an air inlet near the bottom and an air outlet near the top.

[0046] The conical flask has the following dimensions: lower bottom diameter 20 cm, upper bottom diameter 7 cm, and height 26 cm.

[0047] The gas entering the conical flask is filtered through activated carbon. After being filtered through the activated carbon, the gas is split through a Y-shaped tube and enters two conical flasks respectively. The gas flows in the trachea.

[0048] The length, material and diameter of the gas pipes from the Y-type tube to the conical bottles on both sides are the same.

[0049] The activated carbon is connected to the air pump through an air pipe. When the air pump is powered on, the air inlet is opened, the air outlet is connected to the air pipe, and the gas passes into the activated carbon for filtration.

[0050] The method for the mosquito olfactory preference experiment includes the following steps: (1) Connect the equipment and ventilate it to check whether it is completely airtight.

[0051] (2) The gas is pumped into the air pipe by an air pump, filtered through activated carbon, and then diverted to the conical bottles on the left and right sides through a Y-shaped tube. The gas passing through the conical bottle passes through an airflow meter, which is used to control the airflow into the cages on the left and right sides to be the same. The gas is discharged through the circular exhaust port at the top of the middle cube acrylic transparent box.

[0052] (3) Before each experiment, ventilate the equipment for 30 minutes after connecting it to eliminate odor.

[0053] (4) Place the two types of mice into the left and right conical flasks through the top of the conical flask, and then place 100 mated female mosquitoes (3 days old) into the middle cage of the three-cage olfactometer. Start timing.

[0054] (5) Record the number of mosquitoes flying against the wind to the left and right cages 5, 10, 20, and 30 minutes after the start of the experiment.

[0055] (6) After the experiment, ventilate for 30 minutes to remove odor. Disassemble the device and clean it with water.

[0056] The mosquito olfactory preference experiment method is that the conical bottles on both sides can be removed, and the mice can be fixed and placed in the cube-shaped acrylic transparent boxes on the left and right sides to conduct the mosquito olfactory preference experiment.

[0057] In order to explore the selectivity of mosquitoes for different volatile gases of human metabolism, the present invention designed a closed device three-cage olfactometer and conducted a mosquito selection preference experiment. The hair on the back of the mouse was removed, and the same volume of different volatile gas solutions were applied. After the equipment was assembled, the mice applied with different volatile gases were placed in the conical bottles on both sides of the three-cage olfactometer for ventilation. The airflow meter was used to ensure that the airflow into the cages on both sides of the three-cage olfactometer was consistent. The mosquitoes were placed in the middle cage of the three-cage olfactometer and flew against the wind to the cages on the left and right sides to make a choice. The number of mosquitoes that made the choice was counted, and the preference of the mosquitoes was recorded.

[0058] Example 3: Three-cage olfactometer and mosquito olfactory preference experimental method Aedes mosquitoes are important vectors of mosquito-borne infectious diseases. At present, the olfactory signals of Aedes mosquitoes for identifying organisms are still unknown. The present invention mainly proposes a detachable device and experimental method for mosquito olfactory selection preference experiment. It is mainly composed of seven parts, including a cube acrylic transparent box, a cylindrical barrel wall, a conical bottle, a Y-shaped tube, an airflow meter, activated carbon, and an air pump, which are connected by a trachea. It is suitable for exploring the selection preference of mosquitoes for different volatile gases produced by biological metabolism. The air enters the trachea through the air pump, and enters the conical bottle after being filtered by activated carbon and diverted by the Y-shaped tube. Then, it passes through the airflow meter to ensure that the airflow into the cages on the left and right sides is consistent. Female mosquitoes enter from the insect entrance of the middle cage of the three-cage olfactometer, and fly against the wind to the cages on the left and right sides to make a choice. Counting the number of female mosquitoes' choices detects the selection preference of mosquitoes for different volatile gases. The three-cage olfactometer device is easy to disassemble and assemble, occupies a small space, and is very convenient for experimental use; in addition, the device has good airtightness, which can eliminate the influence of the external environment on the experiment. The experimental device is highly flexible and can be disassembled, assembled and modified according to experimental requirements. During the experiment, the experimental results can be seen intuitively, the mosquito counting results are highly reliable, the experiment is easy to operate, repeatable, and the results are highly accurate.

[0059] A three-cage olfactometer comprises a first side cage, a middle cage and a second side cage arranged in sequence, one side of the middle cage is connected with the first side cage through a pipeline, and the other side of the middle cage is connected with the second side cage through a pipeline; the first side cage is connected with a first test mouse container through a pipeline; the second side cage is connected with a second test mouse container through a pipeline, and the first test mouse container and the second test mouse container are respectively connected with an air supply device through a pipeline; an exhaust port is provided on the top of the middle cage.

[0060] A preferred embodiment is as follows: an air flow meter is provided on the pipeline connecting the first side cage and the first test mouse container; an air flow meter is provided on the pipeline connecting the second side cage and the second test mouse container; and the air supply device is an air pump.

[0061] A preferred embodiment is as follows: an air filter is provided on the pipeline connecting the first side cage and the first test mouse container; an air filter is provided on the pipeline connecting the second side cage and the second test mouse container.

[0062] A preferred embodiment is that the air filter is filled with activated carbon.

[0063] A preferred embodiment is that the first test mouse container and the second test mouse container are a conical flask, and the conical flask is provided with an air inlet and an exhaust port.

[0064] An experimental method for testing the olfactory preference of mosquitoes, wherein the experimental device adopts the above-mentioned three-cage olfactometer, and the experimental method comprises the following steps: Step 1: Connect all parts and ventilate to check whether the equipment is completely airtight; Step 2: gas is supplied into the pipeline by the gas supply device, and the gas enters the first test mouse container and the second test mouse container respectively; the gas passing through the first test mouse container and the second test mouse container respectively enters the first side cage and the second side cage through the gas flow meter; the gas is discharged through the exhaust port at the top of the middle cage; Step 3: Before each experiment begins, ventilate all parts after they are connected to eliminate odors.

[0065] Step 4: Two different kinds of test mice are placed into the first test mouse container and the second test mouse container through the upper ports of the first test mouse container and the second test mouse container, respectively, and then the test mosquitoes are placed into the middle cage of the three-cage olfactometer; start timing; Step 5: Record the number of mosquitoes flying against the wind to the first side cage and the second side cage at different times after the start of the experiment; Step 6: After the experiment, ventilate to remove odor.

[0066] The above description is only used to explain the preferred embodiments of the present invention, and is not intended to limit the present invention in any form. Therefore, any modifications or changes made to the present invention under the same inventive spirit should still be included in the scope of protection intended by the present invention.

Claims

1. A three-cage olfactometer, characterized in that: It comprises a first side cage, a middle cage and a second side cage arranged in sequence, one side of the middle cage is connected with the first side cage through a pipeline, and the other side of the middle cage is connected with the second side cage through a pipeline; the first side cage is connected with a first test mouse container through a pipeline; the second side cage is connected with a second test mouse container through a pipeline, and the first test mouse container and the second test mouse container are respectively connected with an air supply device through a pipeline; an exhaust port is provided on the top of the middle cage.

2. The three-cage olfactometer according to claim 1, characterized in that: An air flow meter is arranged on the pipeline connecting the first side cage and the first test mouse container; an air flow meter is arranged on the pipeline connecting the second side cage and the second test mouse container; and the air supply device is an air pump.

3. The three-cage olfactometer according to claim 1, characterized in that: An air filter is provided on the pipeline connecting the first side cage and the first test mouse container; an air filter is provided on the pipeline connecting the second side cage and the second test mouse container.

4. The three-cage olfactometer according to claim 1, characterized in that: The air filter is filled with activated carbon.

5. The three-cage olfactometer according to claim 1, characterized in that: The first test mouse container and the second test mouse container are a conical flask provided with an air inlet and an air outlet.

6. A mosquito olfactory preference experimental method, characterized in that: The experimental device adopts the three-cage olfactometer described in any one of claims 1 to 5, and the experimental method comprises the following steps: Step 1: Connect all parts and ventilate to check whether the equipment is completely airtight; Step 2: gas is supplied into the pipeline by the gas supply device, and the gas enters the first test mouse container and the second test mouse container respectively; the gas passing through the first test mouse container and the second test mouse container respectively enters the first side cage and the second side cage through the gas flow meter; the gas is discharged through the exhaust port at the top of the middle cage; Step 3: Before each experiment, ventilate all parts after they are connected to eliminate odors; Step 4: Two different kinds of test mice are placed into the first test mouse container and the second test mouse container through the upper ports of the first test mouse container and the second test mouse container, respectively, and then the test mosquitoes are placed into the middle cage of the three-cage olfactometer; start timing; Step 5: Record the number of mosquitoes flying against the wind to the first side cage and the second side cage at different times after the start of the experiment; Step 6: After the experiment, ventilate to remove odor.

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