A device for obtaining DNA from the gut environment of juvenile fish

Abstract

The application relates to a young fish stage intestinal environment DNA acquisition device and relates to the field of fish DNA acquisition. The application solves the problems of the existing young fish stage traditional dissection method, i.e. difficulty in acquiring intestinal DNA and waste of samples. The inside of a warehouse body shell is sequentially divided into a dissection warehouse, a flushing warehouse and a sample sampling warehouse from left to right, a first conveying belt is installed in the dissection warehouse, the first conveying belt extends out of the entrance of the dissection warehouse, an operation table is arranged on the first conveying belt, a waste liquid collecting box is installed in the flushing warehouse, a second conveying belt is installed above the waste liquid collecting box, one end of a flushing pipe is connected with buffer liquid installed on the upper end of the lateral wall of the warehouse body shell, the other end of the flushing pipe extends into the flushing warehouse through the warehouse body shell, a third conveying belt is installed in the sample sampling warehouse, collecting pipes are respectively connected with the buffer liquid and sample collecting bottles, and inner sleeve pipes are connected with the collecting pipes extending into the sample sampling warehouse. The application is used for young fish stage intestinal environment DNA acquisition.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of fish DNA acquisition, in particular to a device for acquiring DNA in the intestinal environment of juvenile fish, which is used for extracting DNA in the intestine of juvenile fish. BACKGROUND

[0002] The study of fish diet is an important research content of fish ecology. It mainly includes the analysis of food composition and food selection preference at different stages of fish life history. The juvenile stage usually refers to the stage of small size and incomplete development during the growth of fish. It is also an important growth and development stage of fish in early stage. How is the food composition of fish at this stage, and whether the bait organisms are sufficient, etc. All these information can be obtained by analyzing the food in the intestine, so as to predict the trend of population dynamic change or provide scientific basis for managers to adjust management measures. Therefore, a large number of and long-term monitoring are needed, and food samples need to be quickly acquired.

[0003] The common feature of juvenile fish is small body length, short and thin intestine. For example, the icefish is an annual economic fish. With the continuous pursuit of simple and exquisite life by the public, the demand for high-quality protein provided by fish is increasing. In recent years, due to the breakthrough of stable and high yield technology of icefish in saline-alkali water area in north China, stable high yield and rapid growth of high value indicate that the market demand value and position of icefish in small economic fish are improved. Icefish has the characteristics of persistent juvenile, transparent body, slender body shape and "handsome" appearance. At present, the diet study of icefish still uses the traditional fish dissection method to obtain food samples, and the diet study is based on the microscope technology. This method and related technology are very inconvenient for fish diet monitoring, especially for early juvenile fish. Because the fish body is small (within 4-20 cm), the intestine is short and small (within 3-15 cm), it is difficult to accurately cut with a dissection knife, and it is also difficult to obtain the food inside, so it is difficult to obtain the DNA in the intestinal environment. Usually, skilled technicians need to try several times, which is time-consuming, labor-intensive and sample-wasting.

[0004] In summary, the traditional dissection method used in the juvenile stage has the problems of difficulty in obtaining intestinal DNA and sample waste. SUMMARY

[0005] The purpose of the present application is to solve the problems of difficulty in obtaining intestinal DNA and sample waste in the traditional dissection method used in the juvenile stage, and to provide a device for acquiring DNA in the intestinal environment of juvenile fish.

[0006] The technical scheme of the present application is:

[0007] The application discloses a device for obtaining DNA in the intestinal environment of juvenile fish, which comprises a warehouse body shell, a first conveying belt, an operation table, a waste liquid collecting box, a second conveying belt, a third conveying belt, a flushing pipe, a buffer solution, a collecting pipe, an inner sleeve and a sample collecting bottle; the inside of the warehouse body shell is sequentially divided into an anatomizing warehouse, a flushing warehouse and a sample sampling warehouse from left to right, the first conveying belt is installed in the anatomizing warehouse and extends out of the entrance of the anatomizing warehouse, the operation table is arranged on the first conveying belt, the waste liquid collecting box is installed in the flushing warehouse, the second conveying belt is installed above the waste liquid collecting box, one end of the flushing pipe is connected with the buffer solution installed on the upper end of the lateral wall of the warehouse body shell, and the other end of the flushing pipe extends into the flushing warehouse through the warehouse body shell; the third conveying belt is installed in the sample sampling warehouse, the collecting pipe is connected with the buffer solution and the sample collecting bottle respectively, and the inner sleeve is connected with the collecting pipe extending into the sample sampling warehouse.

[0008] Further, the device further comprises a first partition plate and a second partition plate, the anatomizing warehouse and the flushing warehouse are separated by the first partition plate, and the flushing warehouse and the sample sampling warehouse are separated by the second partition plate.

[0009] Further, the device further comprises a rotating plate and a limiting stopper, the upper end of the rotating plate is rotatably installed at the lower end of the second partition plate, and the limiting stopper is fixedly installed at the lower part of the second partition plate and located at the left side of the rotating plate.

[0010] Further, the upper part of the operation table is provided with an elongated recess, and a plurality of blind holes are formed in the elongated recess at equal intervals along the length direction of the elongated recess.

[0011] Further, the device further comprises an outer sleeve, the outer sleeve comprises a sleeve body, a lotus petal body and a plug-in column, the lotus petal body is coaxially installed at one end of the sleeve body, and the plug-in column is fixedly connected to the lower part of the sleeve body.

[0012] Further, the end of the inner sleeve is conical, and a plurality of through holes are formed in the conical surface.

[0013] Preferably, the device further comprises a bidirectional pump, and the bidirectional pump is installed on the collecting pipe.

[0014] Further, the device further comprises a driving pump, and the driving pump is installed on the flushing pipe.

[0015] Further, the device further comprises a first valve, and the first valve is installed on the collecting pipe connected with the buffer solution.

[0016] Further, the device further comprises a second valve, and the second valve is installed on the flushing pipe.

[0017] Compared with the prior art, the device has the following effects:

[0018] Further, the device further comprises a first partition plate and a second partition plate, the anatomizing warehouse and the flushing warehouse are separated by the first partition plate, and the flushing warehouse and the sample sampling warehouse are separated by the second partition plate.

[0009] Further, the device further comprises a rotating plate and a limiting stopper, the upper end of the rotating plate is rotatably installed at the lower end of the second partition plate, and the limiting stopper is fixedly installed at the lower part of the second partition plate and located at the left side of the rotating plate.

[0010] Further, the upper part of the operation table is provided with an elongated recess, and a plurality of blind holes are formed in the elongated recess at equal intervals along the length direction of the elongated recess.

[0011] Further, the device further comprises an outer sleeve, the outer sleeve comprises a sleeve body, a lotus petal body and a plug-in column, the lotus petal body is coaxially installed at one end of the sleeve body, and the plug-in column is fixedly connected to the lower part of the sleeve body.

[0012] Further, the end of the inner sleeve is conical, and a plurality of through holes are formed in the conical surface.

[0013] Preferably, the device further comprises a bidirectional pump, and the bidirectional pump is installed on the collecting pipe.

[0014] Further, the device further comprises a driving pump, and the driving pump is installed on the flushing pipe.

[0015] Further, the device further comprises a first valve, and the first valve is installed on the collecting pipe connected with the buffer solution.

[0016] Further, the device further comprises a second valve, and the second valve is installed on the flushing pipe.

[0017] Compared with the prior art, the device has the following effects:

[0018] The present application relates to three chambers for obtaining the anatomical compartment of the intestine, the flushing compartment and the food sample obtaining compartment based on environmental DNA research. The expansion flushing of the intestine of the juvenile fish can be realized, and the intestine is sleeved on the outer sleeve and tightened, and the functions of opening the intestine port, flushing the contents and the like are realized. The three chambers of the present application bear different important roles respectively. The anatomical compartment is mainly used for dissecting the fish body and taking off the intestine; the flushing compartment is mainly used for flushing the surface of the intestine to prevent the pollution of the external environmental DNA; the food sample obtaining compartment is mainly used for flushing the contents of the intestine into the collector through the flushing buffer connected by the expansion flushing device (the inner sleeve) to obtain the food sample. The rapid obtaining of the intestinal DNA is realized. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 is the overall structure schematic diagram of the present application; Figure 2 is the sectional view of the operation table 3; Figure 3 is the top view of the operation table; Figure 4 is the partial sectional view of the outer sleeve 16; Figure 5 is the structure schematic diagram of the inner sleeve 10; Figure 6 is Figure 5 is the front view at A.

[0020] In the figure: 1, the compartment body shell, 1-1, the anatomical compartment, 1-2, the flushing compartment, 1-3, the sample obtaining compartment, 2, the first conveying belt, 3, the operation table, 3-1, the long strip-shaped groove, 3-2, the blind hole, 4, the waste liquid collecting box, 5, the second conveying belt, 6, the third conveying belt, 7, the flushing pipe, 8, the buffer solution, 9, the collecting pipe, 10, the inner sleeve, 11, the sample collecting bottle, 12, the first partition plate, 13, the second partition plate, 14, the rotating plate, 15, the limiting stopper, 16, the outer sleeve, 16-1, the sleeve body, 16-2, the lotus petal body, 16-3, the plug-in column, 17, the through hole, 18, the bidirectional pump, 19, the driving pump, 20, the first valve, 21, the second valve. DETAILED DESCRIPTION

[0021] Detailed implementation one: combined with Figures 1 to 6 In this embodiment, the present application includes the compartment body shell 1, the first conveying belt 2, the operation table 3, the waste liquid collecting box 4, the second conveying belt 5, the third conveying belt 6, the flushing pipe 7, the buffer solution 8, the collecting pipe 9, the inner sleeve 10 and the sample collecting bottle 11;

[0022] The inside of the warehouse body shell 1 is divided into the dissection warehouse 1-1, the flushing warehouse 1-2 and the sample sampling warehouse 1-3 from left to right in sequence, the first conveying belt 2 is installed in the dissection warehouse 1-1, and the first conveying belt 2 extends out of the entrance of the dissection warehouse 1-1, the operating table 3 is arranged on the first conveying belt 2, the waste liquid collecting box 4 is installed in the flushing warehouse 1-2, the second conveying belt 5 is installed above the waste liquid collecting box 4, one end of the flushing pipe 7 is connected with the buffer solution 8 installed on the upper end of the lateral wall of the warehouse body shell 1, the other end of the flushing pipe 7 extends into the flushing warehouse 1-2 through the warehouse body shell 1, the third conveying belt 6 is installed in the sample sampling warehouse 1-3, the collecting pipe 9 is connected with the buffer solution 8 and the sample collecting bottle 11 respectively, and the inner sleeve pipe 10 is connected with the collecting pipe 9 extending into the sample sampling warehouse 1-3.

[0023] The fish larvae stage intestinal environment DNA acquisition device of the embodiment mainly realizes through three warehouses, the operating table outside the dissection warehouse is mainly used for basic treatment of the fish larvae, which includes fixing the head of the fish and washing the fish. When the fish larvae on the operating table are conveyed into the inside of the dissection warehouse 1-1 by the first conveying belt 2, the fish intestine is dissected and cut off by the mechanical hand or the operator, the outer sleeve is inserted and installed at both ends of the fish intestine, and is bound and fixed, and the outer sleeve is installed on the operating table, so that the fish intestine is fixed in the length direction.

[0024] In order to avoid pollution, the operating table is moved to the second conveying belt 5 by tilting to the right and up, and the fish intestine is washed through the flushing pipe 7. Since the fish intestine is in an inclined state at this time, the conveying of the second conveying belt 5 can also be controlled according to the actual situation, for example, the second conveying belt 5 is paused until the washing of the outer surface of the fish intestine is completed. The waste liquid generated in the washing process directly flows into the waste liquid collecting box 4, and then the operating table continues to move into the sample sampling warehouse for sample sampling.

[0025] Specific implementation method two: combined with Figure 1 It is explained that the embodiment also includes the first partition plate 12 and the second partition plate 13, the dissection warehouse 1-1 and the flushing warehouse 1-2 are separated by the first partition plate 12, and the flushing warehouse 1-2 and the sample sampling warehouse 1-3 are separated by the second partition plate 13.

[0026] In this way, the first partition plate 12 can block the two warehouses to ensure the environmental hygiene of the operation of the warehouse, and the second partition plate 13 mainly provides a relatively clean sampling environment for the sample sampling warehouse 1-3. The other components and connection relationships are the same as those of the specific implementation method one.

[0027] Specific implementation method three: combined with Figure 1In this embodiment, the rotating plate 14 is rotatably installed at the upper end of the second partition plate 13, and the limiting block 15 is fixedly installed at the lower part of the second partition plate 13 and located at the left side of the rotating plate 14.

[0028] In this way, the rotating plate 14 and the limiting block 15 in combination play the role of a one-way valve, which can ensure the smooth entry of the operation platform into the sample sampling chamber 1-3 and ensure the cleanliness of the sample sampling chamber 1-3. The other components and connection relationships are the same as those in the first or second embodiment.

[0029] Specific embodiment four: combination Figure 2 and Figure 3 In this embodiment, the upper part of the operation platform 3 is provided with an elongated groove 3-1, and a plurality of blind holes 3-2 are processed at equal intervals along the length direction of the elongated groove 3-1.

[0030] In this way, the elongated groove 3-1 is mainly used for placing fish intestines, and the plurality of blind holes 3-2 are used to fix the fish intestines through the sheath 16 on both sides of the fish intestines according to the length of the fish intestines. The other components and connection relationships are the same as those in any one of the first to third embodiments.

[0031] Specific embodiment five: combination Figure 5 and Figure 6 In this embodiment, the sheath 16 includes a sleeve body 16-1, a lotus petal body 16-2, and a plug-in column 16-3. The lotus petal body 16-2 is coaxially installed at one end of the sleeve body 16-1, and the plug-in column 16-3 is fixedly connected to the lower part of the sleeve body 16-1. In this way, the sleeve body 16-1 is used for the entry of the inner sleeve pipe 10 and passes through the lotus petal body 16-2, and the plug-in column 16-3 fixes the fish intestines at the corresponding position. The other components and connection relationships are the same as those in any one of the first to fourth embodiments.

[0032] Specific embodiment six: combination Figure 5 In this embodiment, the end of the inner sleeve pipe 10 is tapered, and a plurality of through holes 17 are processed on the tapered surface. In this way, the through holes 17 can ensure that the buffer solution is quickly injected into the fish intestines. The other components and connection relationships are the same as those in any one of the first to fifth embodiments.

[0033] Specific embodiment seven: combination Figure 1 In this embodiment, the two-way pump 18 is installed on the collection pipe 9.

[0034] In this way, the fish intestine can be injected with buffer solution or the sample in the fish intestine can be collected. The other components and connection relationships are the same as any one of embodiments 1 to 6.

[0035] Embodiment 8: Combination Figure 1 In this embodiment, the embodiment further comprises a driving pump 19, which is installed on the flushing pipe 7. In this way, the pump-out of the buffer solution is powered, and the cleaning of the fish intestine is completed. The other components and connection relationships are the same as any one of embodiments 1 to 7.

[0036] Embodiment 9: Combination Figure 1 In this embodiment, the embodiment further comprises a first valve 20, which is installed on the collection pipe 9 connected with the buffer solution 8. In this way, the collection pipe channel can be opened or closed according to actual needs. The other components and connection relationships are the same as any one of embodiments 1 to 8.

[0037] Embodiment 10: Combination Figure 1 In this embodiment, the embodiment further comprises a second valve 21, which is installed on the flushing pipe 7. In this way, the cleaning of the fish intestine can be controlled at any time. The other components and connection relationships are the same as any one of embodiments 1 to 9.

[0038] Combination Figures 1 to 6 The working principle of the present application is as follows:

[0039] Dissection chamber: The bottom of the dissection chamber has a conveyor belt. A fish is cleaned with deionized water, placed on the conveyor belt, and enters the dissection chamber. The use of the conveyor belt is considered because the present application relates to the dissection of fish with a small body length (within 20 cm in total length). The chamber space is small, and the fish body needs to be cleaned in advance. The conveyor belt has two buckles (prior art) to fix the head and tail of the fish, respectively. The front end of the outer sleeve 16 of the dilatation flusher is inserted into the intestine from the anus of the fish in the dissection chamber. A micro manipulator (prior art) is used to dissect the fish and peel off the skin to expose the complete intestine. The outer sleeve 16 of the dilatation flusher is also inserted into the front end of the intestine. The intestine and the outer sleeve pipe are fixed on the operating table by binding the front and rear ends of the intestine.

[0040] Flushing chamber: The complete intestine obtained is fixed by the dissection chamber and the dilatation flusher, and then enters the flushing chamber through the transmission belt. The micro manipulator is used to flush the outer surface of the intestine with buffer solution, which reduces the DNA pollution in the external environment again.

[0041] Sample collection chamber: the external rinsed intestinal tract enters the food sample collection chamber through the transmission belt. The dilatation flusher is a sleeve type design (referring to the inner sleeve 10 and the outer sleeve 16). The outer sleeve 16 is mainly used for sealing and fixing the intestinal tract and dilatation flushing. The end of the inner sleeve is pen-shaped, which has the function of dilating the intestinal tract. The front end of the inner sleeve has a through hole, which is convenient for the injection of the buffer into the intestinal tract. The end of the inner sleeve is connected with the buffer reservoir, which is compressible, so as to realize the injection pressure of the buffer into the intestinal tract. The end of the inner sleeve is also connected with the sample collector, which is used for collecting the food flushed out of the intestinal tract.

[0042] The above examples are only used to illustrate the technical solutions of the present application, but not to limit it; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that the technical solutions recorded in the foregoing examples can be modified, or some technical features can be replaced by equivalents; 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 application.

Claims

1. A device for obtaining intestinal environment DNA from juvenile fish, characterized in that: It includes a chamber shell (1), a first conveyor belt (2), an operating table (3), a waste liquid collection tank (4), a second conveyor belt (5), a third conveyor belt (6), a rinsing tube (7), a buffer solution (8), a collection tube (9), an inner sleeve (10), a sample collection bottle (11), and an outer sleeve (16). The interior of the outer shell (1) is divided into a dissection chamber (1-1), a rinsing chamber (1-2), and a sample collection chamber (1-3) from left to right. The first conveyor belt (2) is installed inside the dissection chamber (1-1) and extends out of the entrance of the dissection chamber (1-1). The operating table (3) is set on the first conveyor belt (2). The waste liquid collection box (4) is installed inside the rinsing chamber (1-2). The second conveyor belt (5) is installed above the waste liquid collection box (4). One end of the rinsing tube (7) is connected to the buffer solution (8) installed on the upper side of the outer wall of the outer shell (1). The other end of the rinsing tube (7) passes through the outer shell (1) and extends into the rinsing chamber (1-2). The third conveyor belt (6) is installed inside 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) that extends into the sample sampling chamber (1-3); The outer sleeve (16) includes a sleeve body (16-1), a lotus petal body (16-2), and an insert post (16-3). The lotus petal body (16-2) is coaxially mounted at one end of the sleeve body (16-1), and the insert post (16-3) is fixedly connected to the lower part of the sleeve body (16-1). The sleeve body (16-1) is used for the inner sleeve (10) to enter and pass through the lotus petal body (16-2). The end of the inner sleeve (10) is tapered, and multiple through holes (17) are machined on the tapered surface.

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

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

4. The device for obtaining intestinal environment DNA from juvenile fish according to claim 3, characterized in that: The upper part of the operating table (3) is provided with a long strip groove (3-1), and the long strip groove (3-1) is machined with multiple blind holes (3-2) at equal intervals along its length.

5. The device for obtaining intestinal environment DNA from juvenile fish according to claim 4, characterized in that: It also includes a bidirectional pump (18) mounted on the collection pipe (9).

6. The device for obtaining intestinal environment DNA from juvenile fish according to claim 5, characterized in that: It also includes a drive pump (19), which is installed on the flushing pipe (7).

7. A device for obtaining intestinal environment DNA from juvenile fish according to claim 1 or 6, characterized in that: It also includes a first valve (20), which is mounted on a collection tube (9) connected to the buffer solution (8).

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

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