A device for detecting the toxicity of bee pathogens and a method and application of detecting the toxicity of pathogens using the device

Abstract

The present invention relates to a device for detecting the toxicity of bee pathogens and a method and application of detecting the toxicity of pathogens using the device, belonging to the field of bee breeding technology. The present invention provides a device for detecting the toxicity of bee pathogens, comprising a top plate 1, a side plate 2, a partition plate 3, a back plate 4, a front plate 5, a bottom plate 6, a feeding hole 7 and a cavity 8. The device of the present invention can evaluate the impact of the contact mode on the toxicity and transmission speed of bee viruses, providing a basis for the detection of bee viruses in the later stage.

Description

Technical Field

[0001] The present invention relates to the technical field of bee breeding, and in particular to a device for detecting the toxicity of bee pathogens and a method and application of detecting the toxicity of pathogens by using the device. Background Art

[0002] The virulence of a pathogen refers to the invasiveness of the pathogen, that is, the ability of the pathogen to grow, reproduce, spread and diffuse in the body. Some work through bacterial enzymes such as Staphylococcus aureus coagulase, Streptococcus hyaluronidase, and Clostridium perfringens collagenase; some prevent phagocytosis of phagocytes through capsules; and some adhere to host tissues through pili. The virulence of the pathogen is a key indicator for evaluating the transmission speed of the pathogen. Once the bees are infected with the pathogen, the infected bees will be immediately treated according to the virulence and transmission speed of the pathogen, reducing the losses of the bee breeding industry. However, the existing technology is rare in the study of the virulence of bee pathogens. Therefore, there is a lack of a device and method for safely and effectively detecting the virulence of bee pathogens to meet the deficiencies of the existing technology. Summary of the invention

[0003] The purpose of the present invention is to provide a device for detecting the toxicity of bee pathogens and a method and application of detecting the toxicity of pathogens using the device.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0005] The present invention provides a device for detecting the toxicity of bee pathogens, comprising a top plate 1, a side plate 2, a partition plate 3, a back plate 4, a front plate 5, a bottom plate 6, a feeding hole 7 and a cavity 8.

[0006] Preferably, the top plate 1, the side plate 2 and the back plate 4 are plastic plates or wooden plates;

[0007] The partition 3 is a wire mesh;

[0008] The partition 3 divides the cavity 8 into an odd number of chambers;

[0009] The front plate 5 and the bottom plate 6 are plastic plates.

[0010] Preferably, the feeding hole 7 is located on the top plate of the middle chamber;

[0011] Bees carrying pathogens are placed in the middle chamber;

[0012] The feeding hole is circular;

[0013] The diameter of the feeding hole is 1.5 to 2.5 cm.

[0014] Preferably, the pore size of the partition 3 is 0.5-0.8 cm.

[0015] The present invention also provides a method for detecting the toxicity of bee pathogens using the device, comprising the following steps:

[0016] Place the bees carrying the pathogen in the middle chamber, and place healthy bees in other chambers in turn. The infection of the bees in each chamber with the pathogen is tested every 3 to 5 days, and the virulence of the pathogen is determined based on the length of time the bees have been infected with the pathogen.

[0017] Preferably, the density of bees placed in each chamber is 25 to 35 per 300 cm 3 .

[0018] Preferably, the detection method is PCR detection.

[0019] The present invention also provides the use of the device in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens;

[0020] The bee pathogen is wing-removing virus or bee microsporidia.

[0021] The present invention also provides the application of the method described in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens;

[0022] The bee pathogen is wing-removing virus or bee microsporidia.

[0023] The invention provides a device for detecting the toxicity of bee pathogens, and a method and application of detecting the toxicity of pathogens by using the device.

[0024] The device of the present invention is a device for evaluating the toxicity of pathogens in the contact infection mode of bee pathogens. The device of the present invention only sets feeding holes on the top plate of the chamber carrying the pathogen, and the food is only fed to the bees carrying the pathogen. After that, the bees carrying the pathogen pass the food to the bees in other chambers by contact. The PCR detection of the pathogen in the bees is the bees infected with the pathogen. The time from the first bee with the pathogen detected in other chambers to the time when all bees are infected with the pathogen is recorded, which is the speed of the pathogen spreading the pathogen by contact infection. The toxicity of the pathogen is evaluated by the speed of the transmission and the number of dead bees. This method is safe and effective, and provides a scientific basis for the development of the bee industry in the later stage.

[0025] The partition in the device of the present invention is a wire mesh with a certain aperture, and this design provides favorable conditions for bees to contact and infect. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1It is a stereoscopic diagram of the device of the present invention (wherein 1 is a top plate, 2 is a side plate, 3 is a partition plate, 4 is a back plate, 5 is a front plate, 6 is a bottom plate, 7 is a feeding hole, and 9 is a chamber).

[0027] Figure 2 It is a three-dimensional diagram of the device of the present invention (where 8 is the cavity). DETAILED DESCRIPTION

[0028] The present invention provides a device for detecting the toxicity of bee pathogens, comprising a top plate 1, a side plate 2, a partition plate 3, a back plate 4, a front plate 5, a bottom plate 6, a feeding hole 7 and a cavity 8.

[0029] In the present invention, the top plate 1, the side plate 2 and the back plate 4 are plastic plates or wooden plates; the partition 3 is a wire mesh; the partition 3 divides the cavity 8 into an odd number of chambers; the partition 3 is fixed by slots in the back plate, the front plate and the bottom plate; the front plate 5 and the bottom plate 6 are plastic plates; the purpose of setting the plastic plate on the front plate 5 is to facilitate the observation of the state of the bees.

[0030] In the present invention, the feeding hole 7 is located on the top plate of the middle chamber; bees carrying pathogens are placed in the middle chamber; the feeding hole is circular; the diameter of the feeding hole is 1.5 to 2.5 cm, preferably 2 cm.

[0031] In the present invention, the pore size of the separator 3 is 0.5-0.8 cm, preferably 0.65 cm.

[0032] The present invention also provides a method for detecting the toxicity of bee pathogens using the device, comprising the following steps:

[0033] Place the bees carrying the pathogen in the middle chamber, and place healthy bees in other chambers in turn. The infection of the bees in each chamber with the pathogen is tested every 1 day, and the virulence of the pathogen is determined based on the length of time the bees have been infected with the pathogen.

[0034] In the present invention, the density of bees placed in each chamber is 25 to 35 per 300 cm 3 , preferably 30 pieces / 300cm 3 In the present invention, the detection method is PCR detection.

[0035] The present invention also provides the use of the device in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens;

[0036] The bee pathogen is wing-removing virus or bee microsporidia.

[0037] The present invention also provides the application of the method described in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens;

[0038] The bee pathogen is wing-removing virus or bee microsporidia.

[0039] The solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be understood as limiting the protection scope of the present invention.

[0040] The bee species used in the embodiments of the present invention is Apis cerana cerana.

[0041] The preparation method of the honey bee Nosema used in the embodiment of the present invention is: extracting honey bee intestinal tissue and purifying the obtained honey bee Nosema.

[0042] The RT-PCR kit for Nosema orientalis described in the present invention was purchased from Shanghai Kanglang Biotechnology Co., Ltd.

[0043] The detection of the bee wing virus in the present invention is carried out using a kit produced by Beijing Noble Technology Co., Ltd.

[0044] The stereogram of the device of embodiments 1 to 2 of the present invention is as follows Figures 1-2 shown.

[0045] Example 1

[0046] The device is set up as 5 chambers. Each chamber is 10 cm long, 15 cm wide and 20 cm high. The top plate 1, side plate 2, back plate 4, front plate 5 and bottom plate 6 are all made of plastic plates; the partition 3 is a wire mesh with a hole diameter of 0.8 cm; a feeding hole 7 with a diameter of 2.5 cm is opened on the top plate of the middle chamber. 300 newly-released bees are placed in each chamber. The bees in the middle chamber are placed at 10 5 The amount of honey bee microsporidia was fed to obtain bees carrying pathogens. Sugar water was fed to the bees carrying pathogens from the feeding hole of the middle chamber each time. The sugar water was a 50wt% sucrose solution, and the bees obtained the sugar water in a free diet. After feeding the bees with pathogens, the bees in each chamber were tested every 1d for the infection of bee microsporidia. The detection method used the Oriental honey bee microsporidia RT-PCR kit for detection, and the time when the Oriental honey bee microsporidia was detected from other chambers was the 6th day after the bees were fed with pathogens. After testing, the average content of microsporidia in the bees carrying pathogens was 5.6, the average content of microsporidia in the bees in the chamber closest to the middle chamber was 4.3, and the average content of microsporidia in the bees in the outermost chamber was 3.4. This result shows that microsporidia can be transmitted through contact between bees. This result reflects the toxicity of bee microsporidia to a certain extent, and provides a basis for the impact of bee microsporidia on bees in the later stage.

[0047] Example 2

[0048] The device is set up as 5 chambers. Each chamber is 10 cm long, 15 cm wide and 20 cm high. The top plate 1, side plate 2 and back plate 4 are wooden boards, and the front plate 5 and bottom plate 6 are made of plastic boards; the partition 3 is a wire mesh with a hole diameter of 0.5 cm; a feeding hole 7 with a diameter of 1.5 cm is opened on the top plate of the middle chamber. 300 bees that have just left the house are placed in each chamber. The bees in the middle chamber are inoculated with 1×10 bee wing virus positive standard 6 Virus particles were obtained to obtain bees carrying pathogens. Sugar water was fed to the bees carrying pathogens from the feeding hole of the middle chamber each time. The sugar water was a 50wt% sucrose solution, and the bees obtained the sugar water freely. After feeding the bees with pathogens, the bees in each chamber were tested for the infection of the residual wing virus every 1d. The detection method used the bee residual wing virus RT-PCR kit for detection. The time when the bee residual wing virus was detected from other chambers was the 5th day after the bees were fed with pathogens. The result showed that the average content of residual wing virus in the bees in the middle chamber was 6.3×10 5 Virus particles. The average content of residual wing virus in bees close to the middle chamber is 3.2×10 4 Virus particles, the average residual wing virus content in the bees outside the chamber was 2.8×10 4 Virus particles. This result reflects the toxicity of bee stump wings to a certain extent and provides a basis for the impact of bee microsporidia on bees in the later stage.

[0049] The preparation method of the bee wing virus positive standard described in this embodiment is: using bee wing virus DWV RNA as a template, and obtaining it through plasmid construction, extraction, and purification steps.

[0050] It can be seen from the above embodiments that the present invention provides a device for detecting the toxicity of bee pathogens and a method and application of detecting the toxicity of pathogens using the device. The device of the present invention is a device for evaluating the toxicity of pathogens in the contact infection mode of bee pathogens. The device of the present invention only sets feeding holes on the top plate of the chamber carrying the pathogen, and the food is only fed to the bees carrying the pathogen. After that, the bees carrying the pathogen pass the food to the bees in other chambers by contact. The PCR detection of the pathogen in the bees is the bees infected with the pathogen. The time from the detection of the first case of the pathogen in other chambers to the infection of all bees with the pathogen is recorded, that is, the toxicity of the pathogen spread by contact infection, and the toxicity of the pathogen is evaluated by the speed of the transmission. This method is safe and effective, and provides a scientific basis for the development of the bee industry in the future.

[0051] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A device for detecting the toxicity of bee pathogens, characterized in that: It comprises a top plate (1), a side plate (2), a partition plate (3), a back plate (4), a front plate (5), a bottom plate (6), a feeding hole (7) and a cavity (8); The top plate (1), the side plates (2) and the back plate (4) are plastic plates or wooden plates; The partition (3) is a wire mesh; The partition (3) divides the cavity (8) into an odd number of chambers; The front plate (5) and the bottom plate (6) are plastic plates; The feeding hole (7) is located on the top plate of the middle chamber; The middle chamber contains bees carrying pathogens, and the other chambers contain healthy bees. The feeding hole is circular; The diameter of the feeding hole is 1.5-2.5 cm; The pore diameter of the partition (3) is 0.5-0.8 cm.

2. A method for detecting the toxicity of bee pathogens using the device according to claim 1, characterized in that: The steps include: The bees carrying the pathogen are placed in the middle chamber, and healthy bees are placed in other chambers in turn. The infection of the bees in each chamber with the pathogen is tested every 1 day. The virulence and transmission speed of the pathogen are determined based on the length of time the bees are infected with the pathogen, the mortality rate and the number of pathogens.

3. The method according to claim 2, characterized in that The density of bees placed in each chamber is 25-35 / 300cm 3 .

4. The method according to claim 3, characterized in that The detection method is PCR detection.

5. Use of the device according to claim 1 in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens; The bee pathogen is wing-removing virus or bee microsporidia.

6. Use of the method according to any one of claims 2 to 4 in detecting the transmission speed of bee pathogens and / or the toxicity of bee pathogens; The bee pathogen is wing-removing virus or bee microsporidia.

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