Bee hemolymph collection equipment and method

Abstract

The bee hemolymph collecting device comprises a piston core rod, a 0.3 mm capillary tube and a refrigerating machine, the piston core rod is sleeved with a sleeve, a heat preservation cavity is formed between the sleeve and the piston core rod, the bottom of the sleeve is connected with a rotatable capillary tube adapter, and a sealing ring is arranged on the lower portion in the capillary tube adapter; the 0.3 mm capillary tube penetrates through the sealing ring and then integrally penetrates through the capillary tube adapter, a hard water pipe is communicated between the refrigerating machine and the sleeve, and the refrigerating machine is used for pre-cooling refrigerating fluid; comprising the following steps: S1, first-stage hemolymph collection; s2, carrying out second-stage hemolymph collection; s3, collecting the third stage of hemolymph of the bees; s4, collecting the fourth-stage hemolymph of the bees; s5, collecting the fifth-stage hemolymph of the bees. According to the method, a platform for collecting the haemolymph in the full development cycle of the three-type bees is established, the platform is customized according to the specific physiological structure characteristics of the bees in different development stages, and the purpose of ensuring the purity of the extracted haemolymph to the maximum extent is achieved.

Description

Technical Field

[0001] The present invention relates to the field of bee technology, and in particular to bee hemolymph collection equipment and method. Background Art

[0002] Bees play an important role in nature and in the human economy. In nature, the main role of bees is to visit flowers for pollination, and the crop yield and crop quality of more than three-quarters of the world's major crop types depend to some extent on bee pollination.

[0003] The bee's hemolymph circulatory system is different from the closed blood circulatory system of mammals. It is an open circulatory system. In this system, hemolymph flows not only in the heart and major blood vessels, but also in other parts of the bee's body. When it flows outside the blood vessels, it passes through the body cavity, providing nutrition and oxygen to all organs and tissues in the bee's body. It also plays an important role in immunity, wound healing, and defense. As a holometabolous insect, the development of bees usually goes through stages such as the egg stage, larval stage, pupal stage, and adult stage, which makes the collection methods of hemolymph at different developmental stages significantly different.

[0004] During the larval stage, the internal organs of bee larvae gradually develop as they continue to eat and grow. The digestive system, nervous system, muscle system, etc. are all preparing for the lifestyle of adults. When the larvae develop to a certain extent, the adult worker bees will seal the larval cells with wax caps, which marks the end of the larval stage and the beginning of the pupal stage. After the caps are sealed, the larvae will begin to transform into bee pupae, and their bodies will gradually harden in preparation for entering the pupal stage.

[0005] The pupal stage is the transition period from larvae to adults, during which the internal organs of bees undergo significant transformation and differentiation. For example, the digestive system, nervous system, and muscular system of bees will be reorganized and matured at this stage to adapt to the lifestyle of adults.

[0006] The adult stage is the final stage of the bee's life cycle, when the bee is fully developed and capable of reproduction, collecting pollen and nectar, building hives, and protecting the colony. The queen bee is responsible for laying eggs, the drones mainly mate with the queen bee, and the worker bees perform most of the work in the colony, such as collecting food, caring for larvae, and maintaining the hive. Adult bees usually live for a few weeks to a few months, depending on the species of bee and the workload.

[0007] A bee colony is composed of a queen bee, worker bees and drones, and there are different degrees of differences in the development process, organ structure and functional behavior of the three. Therefore, the method for collecting honey bee hemolymph should be tailored according to the specific physiological structure characteristics of its different developmental stages. The purpose of the present invention is to build a collection platform for the hemolymph of honey bees throughout the development cycle of three types of bees to solve the above problems and achieve the purpose of ensuring the purity of the extracted hemolymph to the greatest extent. Summary of the invention

[0008] The purpose of the present invention is to solve the shortcomings in the prior art and to propose a bee hemolymph collection device and method.

[0009] In order to achieve the above object, the present invention adopts the following technical solutions:

[0010] Bee hemolymph collection equipment, including: a piston core rod and a 0.3 mm capillary tube, the piston core rod is sleeved with a sleeve, a heat preservation chamber is provided between the sleeve and the piston core rod, the bottom of the sleeve is connected to a rotatable capillary tube adapter, a sealing ring is provided at the bottom of the capillary tube adapter, and the 0.3 mm capillary tube passes through the sealing ring and then passes through the capillary tube adapter as a whole;

[0011] A refrigerator, wherein a hard water pipe is connected between the refrigerator and the sleeve, and the refrigerator is used for precooling the refrigerant.

[0012] As a further technical solution of the present invention, the capillary adapter keeps the connection between the 0.3 mm capillary and the sleeve sealed by rotation.

[0013] Insert the piston core rod into the sleeve and push it to the bottom. Use a hard water pipe to connect the sleeve and the pre-cooled refrigerant in the refrigerator. When the piston core rod is pulled out, the refrigerant flows into the insulation chamber, and the 0.3mm capillary tube absorbs the hemolymph sample.

[0014] The bee hemolymph collection method comprises the following specific steps:

[0015] S1: First stage hemolymph collection:

[0016] S1a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone larvae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples;

[0017] S1b: Use a blood collection needle to gently puncture the middle part of the larva's back, apply slight pressure to the body to allow hemolymph to extravasate, and use the above-mentioned bee hemolymph collection equipment to aspirate hemolymph;

[0018] S1c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0019] S2: Second stage hemolymph collection:

[0020] S2a: Pretreatment: Wash the larvae with sterile saline and dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples;

[0021] S2b: Use a blood sampling needle to make a small incision in the cuticle of the head area of ​​the larvae, apply slight pressure to the body to allow hemolymph to extravasate, and use the above-mentioned bee hemolymph collection equipment to aspirate hemolymph;

[0022] S2c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0023] S3: Collection of honey bee hemolymph in the third stage:

[0024] S3a: Pretreatment: Wash the bee pupae with sterile saline and then dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples;

[0025] S3b: Turn the back of the worker bee and queen bee pupa upward, adjust the head angle so that the head and body are in a straight line, use a blood sampling needle to pierce the bee pupa head about 1mm above the base of the antenna, gently squeeze the bee pupa chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph; turn the back of the drone bee pupa upward, adjust the head angle so that the head and body are in a straight line, use a blood sampling needle to scratch the compound eyes about 2mm on both sides of the antennae of the bee pupa head, gently squeeze the bee pupa chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph;

[0026] S3c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0027] S4: Collection of honey bee hemolymph in the fourth stage:

[0028] S4a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone pupae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples;

[0029] S4b: inserting a blood sampling needle into the intersegmental membrane of the 3rd to 4th abdominal segments of the honey bee pupa, and using the above-mentioned honey bee hemolymph collection equipment to absorb hemolymph;

[0030] S4c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0031] S5: Collection of hemolymph from the fifth stage of honey bees:

[0032] S5a: Pretreatment: Place the adult bees in a refrigerator at -20℃ for 3-5 minutes for cryoanaesthesia to prevent the adult bees from stinging the experimental operators. After the bees lose their ability to move, use sterile saline to clean the surface of the adult bees and use dust-free paper to absorb the moisture on the surface to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples.

[0033] S5b: Use forceps to remove one antenna of an adult bee from its root, gently squeeze the bee's thorax and abdomen to promote the outflow of hemolymph from the antennae's root, and use the above-mentioned bee hemolymph collection equipment to absorb the hemolymph;

[0034] S5c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for frozen storage.

[0035] As a further technical solution of the present invention, in S1, the queen bees, worker bees and drone larvae processed include 5- and 6-day-old queen bees, worker bees and drones.

[0036] As a further technical solution of the present invention, in S2, the processed larvae include 7-11 day old queen bees, 7-12 day old worker bees, and 7-14 day old drones.

[0037] As a further technical solution of the present invention, in S3, the processed bee pupae include 12-13 day old queen bees, 13-15 day old worker bees, and 15-17 day old drones.

[0038] As a further technical solution of the present invention, in S4, the queen bee, worker bees, and drone pupae processed include 14-16 day old queen bees, 16-21 day old worker bees, and 18-24 day old drones.

[0039] As a further technical solution of the present invention, in said S5, the bees processed include three types of bees in the adult stage.

[0040] The beneficial effects of the present invention are as follows: a collection platform for hemolymph of the entire developmental cycle of honey bees is built, which is tailored according to the specific physiological structure characteristics of honey bees at different developmental stages, so as to achieve the purpose of ensuring the purity of the extracted hemolymph to the greatest extent. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 This is a schematic diagram of the operation of gently piercing the middle part of the back of a larva with a blood collection needle in the method for collecting honey bee hemolymph proposed by the present invention;

[0042] Figure 2 This is a diagram of the internal organs of a bee during the second stage;

[0043] Figure 3 This is a schematic diagram of the operation of using a blood sampling needle to make a small incision on the cuticle of the head area of ​​the larva in the method for collecting honey bee hemolymph proposed by the present invention;

[0044] Figure 4 This is a diagram of the internal organs of a bee in the third stage;

[0045] Figure 5 This is a schematic diagram of the operation of the bee hemolymph collection method proposed by the present invention, using a blood sampling needle to pierce the bee pupa head antennae about 1 mm above the root;

[0046] Figure 6 This is a diagram of the internal organs of a bee in the fourth stage;

[0047] Figure 7 This is a schematic diagram of the operation of inserting a blood sampling needle into the intersegmental membrane of the 3rd and 4th abdominal segments of a honey bee pupa in the honey bee hemolymph collection method proposed by the present invention;

[0048] Figure 8 This is a schematic diagram of the structure of the bee hemolymph collection equipment proposed by the present invention.

[0049] In the figure: 1. piston core rod; 2. sleeve; 3. sealing ring; 4. capillary adapter; 5. 0.3mm capillary; 6. hard water pipe; 7. refrigerator; 8. insulation chamber. DETAILED DESCRIPTION

[0050] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained below in conjunction with specific implementation methods.

[0051] Please see attached Figure 8 , a bee hemolymph collection equipment, comprising a piston core rod 1 and a 0.3 mm capillary tube 5, the piston core rod 1 is sleeved with a sleeve 2, a heat preservation chamber 8 is arranged between the sleeve 2 and the piston core rod 1, the bottom of the sleeve 2 is connected to a rotatable capillary tube adapter 4, a sealing ring 3 is arranged at the bottom of the capillary tube adapter 4, and the 0.3 mm capillary tube 5 passes through the sealing ring 3 and then passes through the capillary tube adapter 4 as a whole;

[0052] A refrigerator 7 is provided, wherein a hard water pipe 6 is connected between the refrigerator 7 and the sleeve 2, and the refrigerator 7 is used for precooling the refrigerant.

[0053] In a preferred embodiment, the capillary adapter 4 keeps the connection between the 0.3 mm capillary 5 and the sleeve 2 sealed by rotation.

[0054] Insert the piston core rod 1 into the sleeve 2 and push it to the bottom, and use the hard water pipe 6 to connect the sleeve 2 and the pre-cooled refrigerant in the refrigerator 7; when the piston core rod 1 is pulled out, the refrigerant flows into the insulation chamber 8, and the 0.3mm capillary 5 absorbs the hemolymph sample.

[0055] The bee hemolymph collection method comprises the following specific steps:

[0056] S1: First stage hemolymph collection:

[0057] S1a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone larvae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples;

[0058] S1b: Use a blood collection needle to gently puncture the middle part of the larva's back, apply slight pressure to the body to allow hemolymph to extravasate, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph, such as Figure 1 As shown;

[0059] S1c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0060] The first stage of queen bee, worker bee and drone larvae includes 5- and 6-day-old queen bees, worker bees and drones. During this period, the larvae's digestive system is not fully developed, and the midgut and hindgut are disconnected. The residues produced after the midgut digests food cannot enter the hindgut and be excreted from the body. The bee larvae will experience rapid growth at this stage, their bodies will gradually become larger, and they will gradually become straighter from the initial C shape. At the same time, they will also shed their skin to support their growth. Moreover, not only are the external changes obvious, but the internal organs of the larvae are also gradually developing, preparing for the next pupal stage.

[0061] S2: Second stage hemolymph collection:

[0062] S2a: Pretreatment: Wash the larvae with sterile saline and dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples;

[0063] S2b: Use a blood collection needle to make a small incision in the cuticle of the larvae's head area and apply slight pressure to the body to allow hemolymph to extravasate. Use the bee hemolymph collection equipment described above to draw out the hemolymph, e.g. Figure 3 As shown;

[0064] S2c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0065] The second stage of larvae includes queen bees aged 7-11 days, worker bees aged 7-12 days, and drones aged 7-14 days. This stage includes not only the late larval stage of bees, but also the pre-pupa stage of bees. During this stage, the internal organs of bees will undergo major transformation and differentiation; for example, glands such as the hypopharyngeal gland, maxillary gland, salivary gland, and organs such as antennae and brain will mature at this stage; the wings and legs of bees begin to appear in the pre-pupa stage, and these appendages will further develop and improve in the pupal stage.

[0066] S3: Collection of honey bee hemolymph in the third stage:

[0067] S3a: Pretreatment: Wash the bee pupae with sterile saline and then dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples;

[0068] S3b: Turn the back of the worker bee and queen bee pupa upward, adjust the head angle so that the head and body are in a straight line, use a blood sampling needle to pierce the bee pupa's head antennae about 1mm above the root, gently squeeze the bee pupa's chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph; turn the back of the drone bee pupa upward, adjust the head angle so that the head and body are in a straight line, use a blood sampling needle to scratch the compound eyes about 2mm on both sides of the bee pupa's head antennae root, gently squeeze the bee pupa's chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph, such as Figure 5 As shown;

[0069] S3c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0070] The third stage of bee pupae include 12-13 day old queen bees, 13-15 day old worker bees, and 15-17 day old drones; during this stage, the bee pupae's eye color will gradually change from white to pink, and then to red; the bee pupae begin to form a new exoskeleton in preparation for the adult stage, and the exoskeleton color will gradually darken from white to light yellow; the bee's internal organs such as the hypopharyngeal glands, maxillary glands, salivary glands and other glands as well as antennae, brain and other organs will further develop and mature.

[0071] S4: Collection of honey bee hemolymph in the fourth stage:

[0072] S4a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone pupae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples;

[0073] S4b: Pierce the blood collection needle into the intersegmental membrane of the 3rd to 4th abdominal segment of the honey bee pupa, and use the above-mentioned honey bee hemolymph collection equipment to absorb hemolymph, such as Figure 7 As shown;

[0074] S4c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation;

[0075] The fourth stage includes 14-16 day old queen bees, 16-21 day old worker bees, and 18-24 day old drones. It is a critical period of metamorphosis and development for bees. At this time, the internal structure and organs of the bees have matured and are preparing to become adults. At the same time, they begin to prepare for emergence, including secreting substances to soften the pupal shell so that they can break out of the pupal shell and become adults.

[0076] S5: Collection of hemolymph from the fifth stage of honey bees:

[0077] S5a: Pretreatment: Place the adult bees in a refrigerator at -20℃ for 3-5 minutes for cryoanaesthesia to prevent the adult bees from stinging the experimental operators. After the bees lose their ability to move, use sterile saline to clean the surface of the adult bees and use dust-free paper to absorb the moisture on the surface to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples.

[0078] S5b: Use forceps to remove one antenna of an adult bee from its root, gently squeeze the bee's thorax and abdomen to promote the outflow of hemolymph from the antennae's root, and use the above-mentioned bee hemolymph collection equipment to absorb the hemolymph;

[0079] S5c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for frozen storage.

[0080] The fifth stage includes the adult period of the three types of bees; the adult stage is the final stage of the bee life cycle, when the bees are fully developed, have the ability to reproduce, and assume various social responsibilities in the hive; the adult's digestive system, nervous system, and reproductive system are all mature and able to perform corresponding physiological functions.

[0081] Operation Notes:

[0082] 1. To ensure the purity of hemolymph and the accuracy of analysis, the experimental equipment should be sterilized before collecting hemolymph.

[0083] 2. In order to reduce oxidation and contamination of hemolymph during collection, hemolymph should be collected quickly and stored in an ice water bath (2-4°C) to prevent melanization.

[0084] 3. After the sample is collected, the hemolymph must be placed in an ultra-low temperature refrigerator at -80°C as soon as possible. This can effectively extend the storage time of the sample and reduce the chemical and biological changes that may occur during storage, thereby ensuring the stability of the hemolymph sample and the reliability of subsequent experimental research.

[0085] 4. Divide the hemolymph into small containers and store them in sealed containers to reduce the area of ​​hemolymph in contact with air and reduce the risk of oxidation. When used later, selectively thaw according to the required number of samples to avoid repeated freezing and thawing.

[0086] From the above description, it can be seen that the above-mentioned embodiments of the present invention achieve the following technical effects: a hemolymph collection platform for the entire developmental cycle of honey bees is built, which is tailored according to the specific physiological structure characteristics of honey bees at different developmental stages, so as to achieve the purpose of ensuring the purity of the extracted hemolymph to the greatest extent.

[0087] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention (including the claims) is limited to these examples. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0088] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. Bee hemolymph collection equipment, characterized in that: include: A piston core rod (1) and a 0.3 mm capillary tube (5), wherein the piston core rod (1) is externally sleeved with a sleeve (2), a heat preservation chamber (8) is provided between the sleeve (2) and the piston core rod (1), the bottom of the sleeve (2) is connected to a rotatable capillary tube adapter (4), a sealing ring (3) is provided at the lower part of the capillary tube adapter (4), and the 0.3 mm capillary tube (5) passes through the sealing ring (3) and then passes through the capillary tube adapter (4) as a whole; A refrigerator (7), wherein a hard water pipe (6) is connected between the refrigerator (7) and the sleeve (2), and the refrigerator (7) is used for precooling the refrigerant.

2. The bee hemolymph collection equipment according to claim 1, characterized in that: The capillary adapter (4) is rotated to keep the connection between the 0.3 mm capillary (5) and the sleeve (2) sealed.

3. A method for collecting honey bee hemolymph, characterized in that: The specific steps include: S1: First stage hemolymph collection: S1a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone larvae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples; S1b: Use a blood collection needle to gently puncture the middle part of the larva's back, apply slight pressure to the body to allow hemolymph to extravasate, and use the above-mentioned bee hemolymph collection equipment to aspirate hemolymph; S1c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation; S2: Second stage hemolymph collection: S2a: Pretreatment: Wash the larvae with sterile saline and dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples; S2b: Use a blood sampling needle to make a small incision in the cuticle of the head area of ​​the larvae, apply slight pressure to the body to allow hemolymph to extravasate, and use the above-mentioned bee hemolymph collection equipment to aspirate hemolymph; S2c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation; S3: Collection of honey bee hemolymph in the third stage: S3a: Pretreatment: Wash the bee pupae with sterile saline and then dry the surface moisture with dust-free paper to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples; S3b: Turn the back of the worker bee and queen bee pupa upward, adjust the head angle so that the head and the body are in a straight line, use a blood sampling needle to pierce the bee pupa head about 1mm above the base of the antenna, gently squeeze the bee pupa chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph; turn the back of the drone bee pupa upward, adjust the head angle so that the head and the body are in a straight line, use a blood sampling needle to scratch the compound eyes about 2mm on both sides of the base of the antenna of the bee pupa head, gently squeeze the bee pupa chest and abdomen, and use the above-mentioned bee hemolymph collection equipment to absorb hemolymph; S3c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation; S4: Collection of honey bee hemolymph in the fourth stage: S4a: Pretreatment: Use sterile saline to clean the queen bee, worker bees, and drone pupae, and then use dust-free paper to absorb the surface moisture to remove bacteria and feed residues on the surface to prevent contamination of hemolymph samples; S4b: insert the blood sampling needle into the intersegmental membrane of the 3rd and 4th abdominal segments of the honey bee pupa, and use the above-mentioned honey bee hemolymph collection equipment to absorb hemolymph; S4c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for freezing and preservation; S5: Collection of hemolymph from the fifth stage of honey bees: S5a: Pretreatment: Place the adult bees in a refrigerator at -20℃ for 3-5 minutes for cryoanaesthesia to prevent the adult bees from stinging the experimental operators. After the bees lose their ability to move, use sterile saline to clean the surface of the adult bees and use dust-free paper to absorb the moisture on the surface to remove bacteria and feed residues on the surface and prevent contamination of hemolymph samples. S5b: Use forceps to remove one antenna of an adult bee from its root, gently squeeze the bee's thorax and abdomen to promote the outflow of hemolymph from the antennae's root, and use the above-mentioned bee hemolymph collection equipment to absorb the hemolymph; S5c: After the collection is completed, the collected hemolymph is placed in an ultra-low temperature refrigerator at -80°C for frozen storage.

4. The method for collecting honey bee hemolymph according to claim 3, characterized in that: In the S1, the queen bee, worker bee and drone larvae processed include 5- and 6-day-old queen bees, worker bees and drones.

5. The method for collecting honey bee hemolymph according to claim 3, characterized in that: In the S2, the larvae processed include 7-11 day old queen bees, 7-12 day old worker bees, and 7-14 day old drone bees.

6. The method for collecting honey bee hemolymph according to claim 3, characterized in that: In S3, the processed bee pupae include 12-13 day old queen bees, 13-15 day old worker bees, and 15-17 day old drones.

7. The method for collecting honey bee hemolymph according to claim 3, characterized in that: In the S4, the queen bee, worker bees and drone pupae processed include 14-16 day old queen bees, 16-21 day old worker bees and 18-24 day old drones.

8. The method for collecting honey bee hemolymph according to claim 3, characterized in that: In said S5, the processed honey bees include three types of bees in the adult stage.

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