Device for efficiently collecting eDNA molecules

Abstract

The invention discloses a device for efficiently collecting eDNA molecules, and relates to the technical field of eDNA, the device comprises a netting, the netting is a barrel-shaped body, an upper net opening of the netting is connected with a traction rope for dragging, a lower net opening of the netting is connected with a water inlet part, the water inlet part is a barrel-shaped body and is used for guiding water flow into the device, and the water inlet part is internally provided with a sundry intercepting net allowing water to pass through; one end, far away from the netting, of the water inlet part is detachably connected with a filtering part, the filtering part comprises a filter body, a filter plate is fixedly connected in the filter body, a plurality of water passing holes are formed in the filter plate, a plurality of filter box grooves are further formed in the filter plate, filter boxes are placed in the filter box grooves, and filter membranes for collecting eDNA molecules are fixed in the filter boxes. By means of the unique design and structure, efficient collection of eDNA is achieved, meanwhile, pollution and loss in the sample treatment process are reduced, and the accuracy and efficiency of biodiversity monitoring are improved.

Description

Technical Field

[0001] The present invention relates to the technical field of eDNA, and in particular to a device for efficiently collecting eDNA molecules. Background Art

[0002] Environmental DNA (eDNA) is the general term for DNA molecules released into the environment by organisms during their life activities. Its sources are mainly secretions, gametes, skin debris, mucus or excrement of organisms, and it is widely present in environmental media such as water, soil, and sediment. eDNA technology refers to a method of directly extracting DNA fragments from environmental samples and then using sequencing technology for qualitative or quantitative analysis. The development of eDNA technology provides a new, convenient and environmentally friendly means for ecological and biodiversity monitoring.

[0003] Currently, fishery resources are declining due to many factors such as habitat destruction, overfishing, climate change, water pollution and invasion of alien species. Fish communities play an important role in aquatic ecosystems. Their structural composition and distribution changes can effectively reflect the stability and health of aquatic ecosystems. Monitoring the diversity and status of rare fish is a key step in determining and implementing conservation and management strategies. Therefore, rapid and accurate assessment of biodiversity and biological resource quantity is of great significance to the protection and restoration of fishery resources. Summary of the invention

[0004] The purpose of the present invention is to provide a device for efficiently collecting eDNA molecules to solve the above-mentioned technical problems existing in the prior art.

[0005] To achieve the above-mentioned purpose, the present invention provides the following scheme: a device for efficiently collecting eDNA molecules, comprising a net, wherein the net is a cylindrical body, the upper net port of which is connected to a traction rope for towing, and the lower net port of which is connected to a water inlet, the water inlet is a cylindrical body, used to guide water flow into the device, the water inlet is provided with a debris interception net that allows water to pass through, the debris interception net is used to intercept debris that may damage the filter membrane; the end of the water inlet away from the net is detachably connected to a filter part, the filter part comprises a filter body, a filter plate is fixedly connected to the filter body, a plurality of water holes are opened on the filter plate, the water holes can prevent the filter membrane from being damaged by excessive water pressure, a plurality of filter box slots are also provided on the filter plate, filter boxes are placed in the filter box slots, and a filter membrane for collecting eDNA molecules is fixed in the filter box.

[0006] As an optional method, the filter box is detachably connected to a filter box cover, and the filter membrane is detachably fixed in the filter box through the filter box cover. The design of using the filter box to load and unload the filter membrane facilitates the rapid replacement of the filter membrane. By dragging it in the water, the water can quickly pass through the filter membrane and directly collect the filter membrane, eliminating the time of active filtration, greatly improving the collection efficiency, and at the same time greatly reducing the possibility of eDNA molecule degradation.

[0007] As an optional manner, the filter membrane is one or more of a glass fiber filter membrane, a polycarbonate filter membrane, a nitrocellulose filter membrane, a cellulose acetate filter membrane, a mixed cellulose ester, and a nylon filter membrane.

[0008] As an optional method, filter membranes with different pore sizes and materials can be selected according to different collection objects.

[0009] As an optional method, a flow meter is connected to the towing rope, which can measure the speed of the device when towing, ensuring the consistency and accuracy of the adoption.

[0010] As an optional method, a limiting net ring is connected to the lower net opening of the net, and the limiting net ring is a reduced diameter structure, with its large open end connected to the net and the diameter of the small open end being smaller than the outer diameter of the water inlet part, and the water inlet part is limited and fixed in the limiting net ring.

[0011] As an optional method, after the water inlet is loaded into the net, the net is fixed tightly to the outer periphery of the water inlet by a clamp. Using the clamp to hold the net tightly to the water inlet can ensure that all water flows into the filter through the water inlet, and when the water flows through the debris interception, the debris that may damage the filter membrane is intercepted; at the same time, the clamp is used for detachable connection, the overall structure is simple, and it is convenient to clean the device later.

[0012] As an optional method, a counterweight is connected to the upper opening of the net through a traction rope. The counterweight can ensure that the device is underwater and ensure the collection efficiency of eDNA.

[0013] As an optional method, a plurality of connection holes are distributed along the circumferential direction on one end of the cylinder body where the water inlet is connected to the filter part, and the filter part is installed on the connection hole through a screw rod. The use of a screw rod for connection can realize the connection between the filter part and the water inlet on the one hand, and on the other hand, the screw rod presses the filter box and the filter box cover from above into the filter box groove to realize the limiting fixation of the filter box and the filter box cover.

[0014] The present invention provides an efficient eDNA molecule collection device with significant technical effects and beneficial effects. First, the device simplifies the operation process and does not need to filter water samples, which effectively reduces the contamination and loss of eDNA during the collection process and improves the sequencing accuracy. Secondly, the device is simple in design and easy to operate, and can collect eDNA molecules quickly and efficiently, reducing the risk of sample degradation and cross-contamination. In addition, the device is highly adaptable and suitable for different water environments. Whether it is a water area with or without water flow, it can be fixed or moved by a counterweight or a towing device to adapt to environmental changes. The detachable design of the filter membrane is easy to replace and protects the eDNA sample from being washed away by water flow. The overall structural design is reasonable, and the net, water inlet and filter work together to effectively capture eDNA molecules while protecting the filter membrane from damage. In summary, the present invention shows significant advantages in improving the accuracy of biodiversity monitoring, ease of operation and environmental adaptability. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. 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 creative work.

[0016] Figure 1 A schematic diagram of the structure of the device for efficiently collecting eDNA molecules of the present invention;

[0017] Figure 2 A side view of the net in the device for efficiently collecting eDNA molecules of the present invention;

[0018] Figure 3 This is a schematic diagram of the structure of the filter part of the device for efficiently collecting eDNA molecules of the present invention;

[0019] Figure 4 A top view of the filter portion of the device for efficiently collecting eDNA molecules of the present invention;

[0020] In the figure: 1. net; 11. flow meter; 12. counterweight; 13. clamp; 14. limit net ring; 2. water inlet; 21. debris interception net; 22. connecting hole; 3. filter part; 31. screw rod; 32. nut; 33. water hole; 34. filter box slot; 35. filter box cover; 36. filter box. DETAILED DESCRIPTION

[0021] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. 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 creative work are within the scope of protection of the present invention.

[0022] eDNA technology plays an increasingly important role in ecological and biodiversity monitoring due to its high efficiency, sensitivity and environmental friendliness. The core of eDNA technology is to use the genetic material in these environmental samples to detect and identify the presence of specific species without directly observing or capturing these organisms.

[0023] At present, the most commonly used method to obtain eDNA from water is active filtration, that is, to allow water to pass through various types of filter membranes by means of vacuum or pressurization. Collecting, transporting, storing, and filtering large amounts of water samples is time-consuming and labor-intensive. eDNA is easily degraded during this process and increases the chance of cross-contamination, resulting in loss and errors in biodiversity information. In order to reduce the pollution and loss during the eDNA collection process and to address the problem of eDNA molecule loss caused by the cumbersome sampling and processing process, the present invention provides a device that can efficiently collect eDNA molecules. Since the invention is easy to operate and does not require filtering water samples, it can efficiently and quickly collect eDNA molecules and reduce the possibility of sample contamination, thereby improving the accuracy during sequencing and providing more possibilities for accurately evaluating the biodiversity of aquatic environments.

[0024] 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.

[0025] Reference Figures 1 to 4 As shown, the present invention provides a device for efficiently collecting eDNA molecules, including a net 1, a water inlet part 2 and a filter part 3.

[0026] Specifically, the net 1 is a net tube structure with openings at both ends, and its upper net opening is connected to a counterweight 12 or a towing device through a traction rope to ensure that the device is underwater, thereby ensuring the collection efficiency of eDNA; a limiting net ring 14 is fixed at the lower net opening of the net 1 through a clamp 13, and the clamp 13 is used to fix the limiting net ring 14 at the lower net opening of the net 1. The limiting net ring 14 is a reduced-diameter structure, with its large open end connected to the net 1 and the caliber of the small open end being smaller than the outer diameter of the water inlet 2; when in use, the water inlet 2 is placed through the upper net opening of the net 1 and is confined in the limiting net ring 14.

[0027] The water inlet 2 is a cylindrical structure, and its function is to guide water flow into the device. Through its structural design, it allows water to flow and contact the filter membrane. The end of the water inlet 2 facing the net 1 is installed with a debris interception net 21, and the other end is used to connect with the filter 3. The debris interception net 21 is located in the water inlet 2 and is used to intercept debris that may damage the filter membrane. There are multiple connection holes 22 evenly distributed along the circumference on the cylinder at one end of the water inlet 2 connected to the filter 3. Correspondingly, the filter 3 has a screw 31 that cooperates with the connection hole 22. The connection between the filter 3 and the water inlet 2 is achieved through the cooperation of the screw 31 and the nut 32.

[0028] The filter part 3 includes a filter body, which is a circular cylindrical structure with a filter plate fixed inside. The filter plate is provided with a number of water holes 33. The filter plate is also provided with a number of filter box slots 34. Filter boxes 36 are placed in the filter box slots 34, and filter membranes are fixed in the filter boxes 36. A number of screw rods 31 are also traversed on the circumference of the filter body. The screw rods 31 penetrate the filter body in a radial direction. Both ends of the screw rods 31 pass through the filter body and pass through the connecting holes 22 on the water inlet part 2, and then are fixedly connected by nuts 32. The use of screw rods 31 for connection can, on the one hand, realize the connection between the filter part 3 and the water inlet part 2. On the other hand, the screw rods 31 press the filter box 36 and the filter box cover 35 from above into the filter box slots 34 to realize the limiting fixation of the filter box 36 and the filter box cover 35. The water holes 33 on the filter plate allow water to flow through, and by sharing the impact of the water flow entering the filter part 3, the filter membrane is protected from being damaged by the water flow.

[0029] In some optional embodiments of the present invention, in order to facilitate the replacement of the filter membrane, the filter membrane can be detachably fixed in the filter box 36. The filter membrane adopts the same shape as the outer shape of the filter box 36, is inserted through the opening at one end of the filter box 36, and is covered with the filter box cover 35 at the insertion end, and the filter membrane is squeezed and fixed to the bottom of the filter box 36 to achieve fixation. When the filter membrane needs to be replaced, the filter box cover 35 can be disassembled to remove the internal filter membrane. The filter box cover 35 and the filter box 36 can be connected by threaded matching, or can be fixed by buckling.

[0030] Before collection, use alcohol to disinfect all parts of the device, put the filter membrane into the filter box 36, cover the filter box cover 35, put it into the filter box slot 34, and then install the filter part 3 at the lower end of the water inlet part 2 and fix it with the screw rod 31 and the nut 32. After the installation is completed, put the whole device in the water and drag it to collect eDNA. After the collection is completed, collect the filter membrane in the filter box 36.

[0031] The filter membrane is fixed by the filter box cover 35, and the filter box cover 35 covers the filter box 36, which can protect the filter membrane and the eDNA sample, so that the eDNA molecules collected in the filter box 36 are not easily washed away by the water flow, and the filter membrane can be quickly replaced.

[0032] In an embodiment of the present invention, the net 1 is used to capture eDNA in water. Through the structural design of the net 1, water is allowed to pass through and eDNA molecules in the water are captured. The net 1 can be set to a conical structure or a reduced diameter structure with one end larger and the other end smaller to improve the capture efficiency.

[0033] In some optional embodiments of the present invention, a tachometer 11 is fixed to the traction rope at the open end of the net 1, and the tachometer 11 can measure the travel speed of the device when towing to ensure the consistency and accuracy of the use.

[0034] In some optional embodiments of the present invention, the filter membrane can be selected from one or more of a glass fiber filter membrane, a polycarbonate filter membrane, a nitrocellulose filter membrane, a cellulose acetate filter membrane, a mixed cellulose ester, and a nylon filter membrane. In water environment applications, after sample collection, the filter membrane method and the precipitation method can be used to concentrate the sample to extract DNA. The filter membrane method can obtain a higher concentration of DNA. Different filter membrane materials have a significant effect on the enrichment effect of eDNA. For example, in a study of the Pearl River Estuary, four different filter membranes (nitrocellulose membrane, cellulose acetate membrane, glass fiber membrane, and polycarbonate membrane) were used for eDNA enrichment, and the results showed that the cellulose acetate membrane obtained the highest eDNA concentration. When extracting eDNA, the combination of different filter membranes and DNA extraction kits will also affect the extraction efficiency and concentration of eDNA. For example, the concentration of eDNA extracted using the Marine Animal Tissue Genomic DNA Kit is higher than that of the DNeasy Blood and Tissue Kit.

[0035] In some optional embodiments of the present invention, the pore size and material of the filter membrane can be selected according to the type of the target sample to be collected; the pore size not only affects the acquisition rate of eDNA, but also affects the filtration speed.

[0036] In practical applications, in waters with currents, a collecting device with a counterweight 12 can be used and placed in the current. The net 1 automatically aligns with the current under the force of the current, and the counterweight 12 serves to fix the device in the current.

[0037] It should be understood that, in actual applications, the traction rope can also be fixed to a stationary object to achieve the same fixing purpose as the counterweight 12, such as tying the traction rope to a stone at the bottom of the water or to a tree on the shore.

[0038] In waters with slow or no water flow, in order to improve the collection efficiency, a towing device can also be used to pull the device forward on the bottom of the water to achieve the purpose of rapid collection. At this time, the counterweight block 12 can help the entire collection device to always be underwater during towing.

[0039] It should also be understood that the connection method between the filter part 3 and the water inlet part 2 is not limited to the matching method of the screw rod 31 and the nut 32 in this embodiment, and can also adopt the form of snap connection, integral threaded connection, clamp connection, etc.

[0040] Compared with the prior art, the embodiments of the present invention at least disclose the following beneficial effects:

[0041] 1. Efficient collection of eDNA molecules: The present invention provides a device that can efficiently collect eDNA molecules in water, which is of great significance for ecological and biodiversity monitoring.

[0042] 2. Reduce pollution and loss; since the device is easy to operate and does not require filtering water samples, it can reduce the contamination and loss of eDNA during the collection process and improve the accuracy of sequencing.

[0043] 3. Improve the accuracy of biodiversity assessments; by reducing sample contamination and loss, the present invention helps to more accurately assess the biodiversity of aquatic environments.

[0044] 4. Easy operation: The design of the device makes the operation easier and eliminates the need for complicated filtering steps, thereby improving the efficiency of eDNA collection.

[0045] 5. Reduce sample degradation and cross-contamination: By reducing the time during water sample collection, transportation, storage and filtration, the risk of eDNA degradation and cross-contamination is reduced.

[0046] 6. Adaptability to different water environments: The device of the present invention is suitable for water areas with or without currents, and can be fixed or moved by a counterweight 12 or a dragging device to adapt to different water environments.

[0047] 7. Replaceable filter membrane design: The filter membrane can be detachably fixed in the filter box 36, which is convenient for replacement and protects the filter membrane and eDNA samples from being washed away by water.

[0048] 8. A variety of filter membrane materials are available for selection: The present invention provides a variety of filter membrane materials for selection, and the most suitable filter membrane can be selected according to different application scenarios and requirements.

[0049] 9. Reasonable structural design: The structural design of the net 1, the water inlet 2 and the filter 3 is reasonable, which can effectively capture the eDNA molecules and protect the filter membrane from being damaged by water flow.

[0050] The details not described in detail in the present invention are all conventional technical means well known to those skilled in the art.

[0051] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside" and "outside" etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0052] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A device for efficiently collecting eDNA molecules, characterized in that: The invention comprises a net (1), wherein the net (1) is a cylindrical body, wherein the upper net opening is connected to a traction rope for towing, and the lower net opening is connected to a water inlet (2), wherein the water inlet (2) is a cylindrical body for guiding water flow into the device, wherein a debris interception net (21) for allowing water to pass through is arranged in the water inlet (2); the end of the water inlet (2) away from the net (1) is detachably connected to a filter (3), wherein the filter (3) comprises a filter body, wherein a filter plate is fixedly connected to the filter body, wherein a plurality of water holes (33) are formed on the filter plate, wherein a plurality of filter box slots (34) are further provided on the filter plate, wherein a filter box (36) is placed in the filter box slot (34), and a filter membrane for collecting eDNA molecules is fixed in the filter box (36).

2. The device for efficiently collecting eDNA molecules according to claim 1, characterized in that: The filter box (36) is detachably connected to a filter box cover (35), and the filter membrane is detachably fixed in the filter box (36) via the filter box cover (35).

3. The device for efficiently collecting eDNA molecules according to claim 1, characterized in that: The filter membrane is one or more of a glass fiber filter membrane, a polycarbonate filter membrane, a nitrocellulose filter membrane, a cellulose acetate filter membrane, a mixed cellulose ester, and a nylon filter membrane.

4. The device for efficiently collecting cDNA molecules according to claim 3, characterized in that: Depending on the collection object, select filter membranes with different pore sizes and materials.

5. The device for efficiently collecting cDNA molecules according to claim 1, characterized in that: The traction rope is connected to a flow meter (11).

6. The device for efficiently collecting cDNA molecules according to claim 1, characterized in that: The lower mesh opening of the net (1) is connected to a limited mesh ring (14); the limited mesh ring (14) is a reduced diameter structure, with a large opening end connected to the net (1) and a small opening end having a diameter smaller than the outer diameter of the water inlet (2); the water inlet (2) is limited and fixed in the limited mesh ring (14).

7. The device for efficiently collecting cDNA molecules according to claim 6, characterized in that: After the water inlet portion (2) is installed in the net (1), the net (1) is tightly fixed to the outer periphery of the water inlet portion (2) via a clamp (13).

8. The device for efficiently collecting eDNA molecules according to claim 1, characterized in that: The net opening of the net garment (1) is connected to a counterweight block (12) via a traction rope.

9. The device for efficiently collecting cDNA molecules according to claim 1, characterized in that: A plurality of connection holes (22) are distributed along the circumferential direction on one end of the cylinder body where the water inlet portion (2) is connected to the filter portion (3); the filter portion (3) is mounted on the connection hole (22) via a screw rod (31); the screw rod (31) presses the filter box (36) and the filter box cover (35) against the filter box groove (34) from above.

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