Device for acquiring DNA (deoxyribonucleic acid) of intestinal environment in juvenile fish stage

Abstract

The invention discloses a juvenile fish stage intestinal environment DNA (deoxyribonucleic acid) acquisition device, and relates to the field of fish DNA acquisition. According to the method, the problems that intestinal tract DNA is difficult to obtain and samples are wasted in the conventional dissection method adopted in the juvenile fish stage are solved. The interior of a bin shell is sequentially divided into a dissecting bin, a flushing bin and a sample sampling bin from left to right, a first conveying belt is installed in the dissecting bin and extends out of an inlet of the dissecting bin, an operation table is arranged on the first conveying belt, a waste liquid collecting box is installed in the flushing bin, and a second conveying belt is installed above the waste liquid collecting box; one end of the flushing pipe is connected with a buffer solution mounted at the upper end of the outer side wall of the bin body shell, and the other end of the flushing pipe penetrates through the bin body shell and then extends into the flushing bin; the third conveying belt is installed in the sample sampling bin, the collecting pipe is connected with the buffer solution and the sample collecting bottle, and the inner sleeve is connected with the collecting pipe extending into the sample sampling bin. The method is used for obtaining the intestinal environment DNA in the juvenile fish stage.

Description

Technical Field

[0001] The present invention relates to the field of fish DNA acquisition, and particularly to a device for obtaining environmental DNA in the intestine at the juvenile fish stage, which is used to extract DNA from the intestine of juvenile fish. Background Art

[0002] The study of fish feeding habits is an important research content in fish ecology. It mainly includes the analysis of the food composition and food selection preferences at different stages of the fish life history. The juvenile fish stage usually refers to the stage in the growth process of fish when the body size is small and not fully developed. It is also an important early growth and development stage of fish. Information such as the food composition at this stage and whether the bait organisms are sufficient can be obtained through the analysis of the food in the intestine, so as to predict the dynamic change trend of the population quantity or provide a scientific basis for managers to adjust management measures. Therefore, a large amount of long-term monitoring is required, and at the same time, food samples need to be obtained quickly.

[0003] The general characteristics of fish at the juvenile fish stage are small body length, short and thin intestines. For example, Protosalanx chinensis is an annual economic fish. With the current continuous pursuit of a simple and delicate life by the public, fish that provide high-quality protein are increasingly favored by the public. In recent years, due to the breakthrough in the continuous stable production technology of Protosalanx chinensis in the northern saline-alkali waters, the stable high yield and rapid growth of high output value indicate the improvement of the market demand value and status of Protosalanx chinensis among small economic fish. Protosalanx chinensis has continuous juvenile characteristics, is transparent throughout, has a slender body shape, and has a "delicate" appearance. At present, the feeding habit research of Protosalanx chinensis still uses the traditional fish dissection method to obtain food samples and conducts feeding habit research based on microscopic examination technology. This method and related technologies are very inconvenient for monitoring the feeding habits of fish, especially juvenile fish in the early development stage. Due to the small size of the fish body (within 4 - 20 cm) and the short and small intestine (within 3 - 15 cm), it is difficult for a dissecting knife to cut precisely and it is also difficult to obtain the food inside, so it is very difficult to obtain DNA in the intestinal environment. Usually, it requires skilled technicians to try many times, which is time-consuming, laborious and wastes samples.

[0004] In summary, the traditional dissection method used at the juvenile fish stage has problems such as difficulty in obtaining intestinal DNA and wasting samples. Summary of the Invention

[0005] The purpose of the present invention is to solve the problems that the traditional dissection method used at the juvenile fish stage has difficulty in obtaining intestinal DNA and wastes samples, and further provide a device for obtaining environmental DNA in the intestine at the juvenile fish stage.

[0006] The technical solution of the present invention is as follows:

[0007] An intestinal environmental DNA acquisition device for the juvenile fish stage, comprising a housing shell, a first conveyor belt, an operating table, a waste liquid collection box, a second conveyor belt, a third conveyor belt, a flushing pipe, a buffer solution, a collection pipe, an inner sleeve and a sample collection bottle; the interior of the housing shell is divided into an anatomical chamber, a flushing chamber and a sample sampling chamber from left to right in sequence. The first conveyor belt is installed in the anatomical chamber, and the first conveyor belt extends out of the entrance of the anatomical chamber. The operating table is arranged on the first conveyor belt. The waste liquid collection box is installed in the flushing chamber. The second conveyor belt is installed above the waste liquid collection box. One end of the flushing pipe is connected to the buffer solution installed at the upper end of the outer side wall of the housing shell, and the other end of the flushing pipe passes through the housing shell and extends into the flushing chamber. The third conveyor belt is installed in the sample sampling chamber. The collection pipe is respectively connected to the buffer solution and the sample collection bottle. The inner sleeve is connected to the collection pipe extending into the sample sampling chamber.

[0008] Furthermore, it also includes a first partition board and a second partition board. The anatomical chamber and the flushing chamber are separated by the first partition board, and the flushing chamber and the sample sampling chamber are separated by the second partition board.

[0009] Furthermore, it also includes a rotating plate and a limiting block. The upper end of the rotating plate is rotatably installed at the lower end of the second partition board. The limiting block is fixedly installed at the lower part of the second partition board, and the limiting block is located on the left side of the rotating plate.

[0010] Even further, a long strip-shaped groove is provided on the upper part of the operating table, and a plurality of blind holes are processed at equal intervals along the length direction of the long strip-shaped groove.

[0011] Furthermore, it also includes an outer sleeve. The outer sleeve includes a sleeve body, a lotus petal body and an insertion column. The lotus petal body is coaxially installed at one end of the sleeve body, and the insertion column is fixedly connected to the lower part of the sleeve body.

[0012] Furthermore, the end of the inner sleeve is conical, and a plurality of through holes are processed on the conical surface.

[0013] Preferably, it also includes a two-way pump. The two-way pump is installed on the collection pipe.

[0014] Furthermore, it also includes a driving pump. The driving pump is installed on the flushing pipe.

[0015] Furthermore, it also includes a first valve. The first valve is installed on the collection pipe connected to the buffer solution.

[0016] Furthermore, it also includes a second valve. The second valve is installed on the flushing pipe.

[0017] The present invention has the following effects compared with the prior art:

[0018] The present invention relates to three chambers, namely an anatomical chamber for obtaining the intestine, a rinsing chamber, and a food sample obtaining chamber for environmental DNA research. It can expand and rinse the intestine of juvenile fish, tie the intestine on the outer sleeve, and also has functions such as opening the intestinal port and rinsing the contents. The three chambers of the present invention play different important roles respectively. The anatomical chamber is mainly used for dissecting the fish body and removing the intestine; the rinsing chamber is mainly used for rinsing the intestinal surface to prevent the pollution of foreign environmental DNA; the food sample obtaining chamber is mainly used for rinsing the intestinal contents into the collector through the rinsing buffer solution (referring to the inner sleeve) connected to the expansion rinser to obtain food samples. It realizes the rapid acquisition of intestinal DNA. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is the overall structural schematic diagram of the present invention; Figure 2 is a cross-sectional view of the operating table 3; Figure 3 is a top view of the operating table; Figure 4 is a partial cross-sectional view of the outer sleeve 16; Figure 5 is the structural schematic diagram of the inner sleeve 10; Figure 6 is Figure 5 a view from the direction at A.

[0020] In the figure: 1, housing shell; 1-1, anatomical chamber; 1-2, rinsing chamber; 1-3, sample sampling chamber; 2, first conveyor belt; 3, operating table; 3-1, long groove; 3-2, blind hole; 4, waste liquid collection box; 5, second conveyor belt; 6, third conveyor belt; 7, rinsing pipe; 8, buffer solution; 9, collection pipe; 10, inner sleeve; 11, sample collection bottle; 12, first partition board; 13, second partition board; 14, rotating plate; 15, limit stop block; 16, outer sleeve; 16-1, sleeve body; 16-2, lotus petal body; 16-3, insertion column; 17, through hole; 18, two-way pump; 19, driving pump; 20, first valve; 21, second valve. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] Detailed Embodiment 1: In combination with Figures 1 to 6 This embodiment is described. This embodiment includes a housing shell 1, a first conveyor belt 2, an operating table 3, a waste liquid collection box 4, a second conveyor belt 5, a third conveyor belt 6, a rinsing pipe 7, a buffer solution 8, a collection pipe 9, an inner sleeve 10, and a sample collection bottle 11;

[0022] Inside the outer shell 1 of the bin body, it is sequentially divided into an anatomical bin 1-1, a flushing bin 1-2, and a sample sampling bin 1-3 from left to right. The first conveyor belt 2 is installed inside the anatomical bin 1-1, and the first conveyor belt 2 extends out of the entrance of the anatomical bin 1-1. The operating table 3 is arranged on the first conveyor belt 2. The waste liquid collection box 4 is installed inside the flushing bin 1-2. The second conveyor belt 5 is installed above the waste liquid collection box 4. One end of the flushing pipe 7 is connected to the buffer liquid 8 installed at the upper end of the outer side wall of the bin body shell 1, and the other end of the flushing pipe 7 passes through the bin body shell 1 and then extends into the flushing bin 1-2; the third conveyor belt 6 is installed inside the sample sampling bin 1-3. The collection pipe 9 is respectively connected to the buffer liquid 8 and the sample collection bottle 11. The inner sleeve 10 is connected to the collection pipe 9 extending into the sample sampling bin 1-3.

[0023] The device for obtaining intestinal environmental DNA at the juvenile fish stage of this embodiment mainly realizes it through three chambers. Among them, on the operating table outside the anatomical chamber, it is mainly used for basic processing of juvenile fish, including fixing the fish head and cleaning the fish. When the juvenile fish on the operating table is conveyed by the first conveyor belt 2 and enters the inside of the anatomical bin 1-1, the fish intestine is anatomically cut by a manipulator or an operator, and the outer sleeve is inserted at both ends of the fish intestine and tied and fixed. The outer sleeve is installed on the operating table to realize the fixation of the fish intestine in the length direction.

[0024] To avoid contamination, the operating table moves obliquely upward to the right onto the second conveyor belt 5, and the fish intestine is cleaned through the flushing pipe 7. Since the fish intestine is in an inclined state at this time, the conveying situation of the second conveyor belt 5 can also be controlled according to the actual situation. For example, the second conveyor belt 5 is paused until the cleaning work on the outer surface of the fish intestine is completed. The waste liquid generated during the cleaning process directly flows into the waste liquid collection box 4, and then the operating table continues to move and enters the sample sampling bin for sample sampling.

[0025] Specific Embodiment Two: Combined Figure 1 This embodiment is described. This embodiment further includes a first partition 12 and a second partition 13. The anatomical bin 1-1 and the flushing bin 1-2 are separated by the first partition 12, and the flushing bin 1-2 and the sample sampling bin 1-3 are separated by the second partition 13.

[0026] With such a setting, the first partition 12 can block the two chambers to ensure the environmental hygiene of the operations in the chambers. The second partition 13 mainly provides a relatively clean sampling environment for the sample sampling bin 1-3. Other compositions and connection relationships are the same as those in Specific Embodiment One.

[0027] Specific Embodiment Three: Combined Figure 1Regarding this embodiment, this embodiment further includes a rotating plate 14 and a limiting stop 15. The upper end of the rotating plate 14 is rotatably installed at the lower end of the second partition plate 13. The limiting stop 15 is fixedly installed at the lower part of the second partition plate 13, and the limiting stop 15 is located on the left side of the rotating plate 14.

[0028] With such a setting, after the combination of the rotating plate 14 and the limiting stop 15, it functions as a one-way valve, which can not only ensure the smooth entry of the operating table into the sample sampling chamber 1-3, but also ensure the cleanliness of the sample sampling chamber 1-3. Other components and connection relationships are the same as those in the first or second specific embodiment.

[0029] Specific embodiment four: Combining Figure 2 and Figure 3 Regarding this embodiment, a long strip-shaped groove 3-1 is provided at the upper part of the operating table 3 in this embodiment. A plurality of blind holes 3-2 are processed at equal intervals along the length direction of the long strip-shaped groove 3-1.

[0030] With such a setting, the long strip-shaped groove 3-1 is mainly used for placing fish intestines, and for the plurality of blind holes 3-2, different positions are selected according to the length of the fish intestines to fix the fish intestines through the outer sleeves 16 on both sides of the fish intestines. Other components and connection relationships are the same as any one of the first to third specific embodiments.

[0031] Specific embodiment five: Combining Figure 5 and Figure 6 Regarding this embodiment, this embodiment further includes an outer sleeve 16. The outer sleeve 16 includes a sleeve body 16-1, a lotus petal body 16-2, and an insertion column 16-3. The lotus petal body 16-2 is coaxially installed at one end of the sleeve body 16-1, and the insertion column 16-3 is fixedly connected to the lower part of the sleeve body 16-1. With such a setting, the sleeve body 16-1 is used for the inner sleeve 10 to enter and pass through the lotus petal body 16-2, and the insertion column 16-3 fixes the fish intestines in the corresponding position. Other components and connection relationships are the same as any one of the first to fourth specific embodiments.

[0032] Specific embodiment six: Combining Figure 5 Regarding this embodiment, the end of the inner sleeve 10 in this embodiment is conical, and a plurality of through holes 17 are processed on this conical surface. With such a setting, the through holes 17 can ensure the rapid injection of the buffer solution into the fish intestines. Other components and connection relationships are the same as any one of the first to fifth specific embodiments.

[0033] Specific embodiment seven: Combining Figure 1 Regarding this embodiment, this embodiment further includes a two-way pump 18, and the two-way pump 18 is installed on the collecting pipe 9.

[0034] Such a setting facilitates controlling whether to inject the buffer solution into the fish intestine or collect the sample inside the fish intestine. The other components and connection relationships are the same as any one of the first to sixth specific embodiments.

[0035] Specific Embodiment Eight: Figure 1 In describing this specific embodiment, this specific embodiment further includes a driving pump 19, and the driving pump 19 is installed on the flushing pipe 7. Such a setting facilitates providing power for pumping out the buffer solution to complete the cleaning of the fish intestine. The other components and connection relationships are the same as any one of the first to seventh specific embodiments.

[0036] Specific Embodiment Nine: Figure 1 In describing this specific embodiment, this specific embodiment further includes a first valve 20, and the first valve 20 is installed on the collecting pipe 9 connected to the buffer solution 8. Such a setting facilitates opening or closing the collecting pipe channel according to actual needs. The other components and connection relationships are the same as any one of the first to eighth specific embodiments.

[0037] Specific Embodiment Ten: Figure 1 In describing this specific embodiment, this specific embodiment further includes a second valve 21, and the second valve 21 is installed on the flushing pipe 7. Such a setting facilitates controlling at any time whether to clean the fish intestine or not. The other components and connection relationships are the same as any one of the first to ninth specific embodiments.

[0038] Combined with Figures 1 to 6 Describe the working principle of the present invention:

[0039] Dissection Chamber: There is a conveyor belt at the bottom of the dissection chamber. A fish is cleaned with deionized water and placed on the conveyor belt and enters the dissection chamber. The way of using the conveyor belt is considered because the present invention is related to the dissection of small-sized fish (total length within 20 cm). The chamber space itself is not large, and the fish body needs to be cleaned in advance. There are two buckles (prior art) on the conveyor belt to fix the head and tail of the fish respectively. In the dissection chamber, the front end of the outer sleeve 16 in the expansion flusher is inserted into the intestine from the anus of the fish body. Operate the micro manipulator (prior art) to dissect the fish body, peel off the skin and flesh to expose the complete intestine. The outer sleeve 16 in the expansion flusher is also inserted into the front end of the intestine. And the intestine and the outer sleeve pipe are fixed on the operating table by tying them at the front and rear ends of the intestine.

[0040] Flushing Chamber: The obtained complete intestine is fixed with the expansion flusher in the dissection chamber and then enters the flushing chamber through the conveyor belt. Operate the micro manipulator to wash the outer surface of the intestine in detail with the buffer solution to further reduce the DNA contamination in the external environment.

[0041] Sample acquisition chamber: The externally rinsed intestine enters the diet sample acquisition chamber through the conveyor belt. The expansion rinser has a sleeve design (referring to the inner sleeve 10 and the outer sleeve 16). The outer sleeve 16 is mainly used for sealing and fixing the intestine and for expansion rinsing. The end of the inner sleeve is pen-point shaped and has the function of expanding the intestine. There are through holes at the front end of the inner sleeve to facilitate the injection of the buffer solution into the intestine. The end of the inner sleeve is connected to a buffer solution reservoir, which is compressible, so as to realize injecting the buffer solution into the intestine under the injection pressure. The end of the inner sleeve is also connected to a sample collector for collecting the food rinsed out of the intestine.

[0042] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A device for obtaining DNA from the intestinal environment of juvenile fish, characterized by: It comprises a warehouse shell (1), a first conveyor belt (2), an operating table (3), a waste liquid collection box (4), a second conveyor belt (5), a third conveyor belt (6), a flushing tube (7), a buffer solution (8), a collection tube (9), an inner sleeve (10) and a sample collection bottle (11); The interior of the chamber shell (1) is divided into a dissecting chamber (1-1), a flushing chamber (1-2) and a sample sampling chamber (1-3) from left to right. The first conveyor belt (2) is installed in the dissecting chamber (1-1), and the first conveyor belt (2) extends out of the entrance of the dissecting chamber (1-1). The operating table (3) is arranged on the first conveyor belt (2). The waste liquid collection box (4) is installed in the flushing chamber (1-2). The second conveyor belt (5) is installed above the waste liquid collection box (4). One end of the flushing pipe (7) is connected to a buffer solution (8) installed at the upper end of the outer wall of the chamber shell (1), and the other end of the flushing pipe (7) passes through the chamber shell (1) and extends into the flushing chamber (1-2). The third conveyor belt (6) is installed in the sample sampling chamber (1-3), the collection tube (9) is connected to the buffer solution (8) and the sample collection bottle (11) respectively, and the inner sleeve (10) is connected to the collection tube (9) extending into the sample sampling chamber (1-3).

2. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 1, characterized in that: It also includes a first partition (12) and a second partition (13); the dissection chamber (1-1) and the flushing chamber (1-2) are separated by the first partition (12); and the flushing chamber (1-2) and the sample sampling chamber (1-3) are separated by the second partition (13).

3. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 2, characterized in that: It also includes a rotating plate (14) and a limit stopper (15), wherein the upper end of the rotating plate (14) is rotatably mounted on the lower end of the second partition plate (13), the limit stopper (15) is fixedly mounted on the lower part of the second partition plate (13), and the limit stopper (15) is located on the left side of the rotating plate (14).

4. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 3, characterized in that: A long strip groove (3-1) is provided on the upper part of the operating table (3), and a plurality of blind holes (3-2) are processed at equal intervals along the length direction of the long strip groove (3-1).

5. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 4, characterized in that: The outer sleeve (16) further comprises a sleeve body (16-1), a lotus petal body (16-2) and an inserting column (16-3), wherein the lotus petal body (16-2) is coaxially mounted on one end of the sleeve body (16-1), and the inserting column (16-3) is fixedly connected to the lower part of the sleeve body (16-1).

6. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 5, characterized in that: The end of the inner sleeve (10) is tapered, and a plurality of through holes (17) are machined on the tapered surface.

7. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 6, characterized in that: It also includes a bidirectional pump (18) which is installed on the collecting pipe (9).

8. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 7, characterized in that: It also includes a driving pump (19), and the driving pump (19) is installed on the flushing pipe (7).

9. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 1 or 8, characterized in that: It also includes a first valve (20), which is installed on a collection tube (9) connected to the buffer solution (8).

10. The device for acquiring DNA from the intestinal environment of juvenile fish according to claim 9, characterized in that: It also includes a second valve (21), which is mounted on the flushing pipe (7).

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