Minimally invasive accurate sampling method for sea cucumber coelomic fluid

Abstract

The invention relates to the technical field of marine organism sampling, in particular to a minimally invasive accurate sampling method for sea cucumber coelomic fluid. The method comprises the following steps: scanning a sea cucumber body cavity by using an ultrasonic detector, and selecting a position which is thin in back midline body wall and avoids viscera as a sampling part; an injector provided with a special double-layer sleeve type sampling needle is selected, obliquely and slowly pierces into the body wall by 3-6 mm at an angle of 45 degrees and stretches into the body cavity, and the body cavity fluid of the sea cucumber is slowly sucked. In the sampling process, the exposure time of the sea cucumbers is controlled within 1 minute, so that the stimulation to the sea cucumbers is reduced. The minimally invasive accurate sampling method provided by the invention is simple and convenient to operate, rapid and accurate collection of the sea cucumber coelomic fluid can be realized, and the problem of sample pollution caused by contact of the coelomic fluid with dissecting tools and vessels in the sampling process can be effectively prevented. In addition, the wound caused by the method is tiny, the damage to the sea cucumbers is small, the sea cucumbers can recover quickly after sampling, and the rare sea cucumber resources can be effectively protected while scientific research is developed.

Description

Technical Field

[0001] The invention relates to the technical field of marine biological sampling, and in particular to a method for accurately sampling sea cucumber coelomic fluid with minimal trauma. Background Art

[0002] The ecological function of tropical coral reef sea cucumbers is very important, but many wild sea cucumber species are endangered and their resources are in urgent need of protection. Scientific and effective protection requires revealing their basic physiological and ecological characteristics, and scientific research and sampling are very necessary. In addition, as a marine organism with important ecological and economic value, sea cucumbers contain rich bioactive substances and physiological information in their coelomic fluids. For example, the large number of coelomocytes and hormones contained in the coelomic fluid can reflect the status of a variety of physiological indicators. Therefore, coelomic fluid is the preferred sample for studying sea cucumber physiological indicators. Because there are many tissues and organs inside the body cavity of sea cucumbers, the commonly used needle puncture method for obtaining coelomic fluid will be blocked by internal tissues, resulting in sampling failure, and only traditional anatomical destructive sampling methods can be used. At present, there are many problems with traditional sampling methods when obtaining sea cucumber coelomic fluid.

[0003] Traditional coelomic fluid sampling generally adopts the method of anatomical sampling. The body wall of the sea cucumber is cut open from the anus to the mouth with a scalpel to collect the coelomic fluid.

[0004] However, the traditional method of body cavity fluid sampling has the following disadvantages:

[0005] 1. Damage to experimental animals is not conducive to the protection of sea cucumber resources. Most existing sampling methods are invasive, which may cause serious damage to sea cucumbers and even lead to their death. This not only affects the subsequent research and observation of individual sea cucumbers, but is also not conducive to the sustainable use of sea cucumber resources. For rare species of sea cucumbers, this damage may bring irreparable losses.

[0006] 2. Traditional sampling methods are complicated to operate, which increases the cost and difficulty of sampling; for larger sea cucumbers, the body wall is thick and the sampling area is large, so the sampling process is time-consuming and laborious. This also limits the efficiency of obtaining sea cucumber coelomic fluid in large-scale research and practical applications.

[0007] 3. Invasive sampling may introduce external contaminants, affecting the analysis results of the body cavity fluid. There are a large number of bacteria on the surface of sea cucumbers and in the external environment. During the dissection process, the body cavity fluid may be contaminated by sampling tools and utensils. In addition, the dissection sampling time is long, and the biologically active substances in the sea cucumber body cavity fluid are easily oxidized and deteriorated when exposed to air.

[0008] In summary, the existing methods for sampling the coelomic fluid of sea cucumbers have problems such as damaging sea cucumbers, easy introduction of contamination, and complex operation. Therefore, it is very necessary to develop a minimally invasive coelomic fluid sampling method suitable for sea cucumbers, which contain a large amount of tissue in their body cavity. This method can reveal the physiological state of sea cucumbers without killing them, achieve animal welfare protection, and effectively protect the rare and endangered sea cucumber resources that are about to be depleted. Summary of the invention

[0009] In view of this, the present invention provides a minimally invasive and precise sampling method for sea cucumber coelomic fluid, which can achieve slight trauma (recoverable), is easy to operate, and has high efficiency. It also overcomes the blind method for accurately collecting coelomic fluid and avoids the adverse effects of traditional anatomical sampling.

[0010] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0011] The present invention provides a minimally invasive sampling device for sea cucumber coelomic fluid, comprising a sampling tube, a needle connector 5, a syringe 2, a piston 3, and a push rod 1;

[0012] The sampling tube comprises an outer tube 7 and an inner tube 6, wherein the outer tube is sleeved outside the inner tube, and the same end of the outer tube and the inner tube is connected to the needle connector;

[0013] The outer tube is a side hole needle having at least two side holes 8;

[0014] The syringe is provided with an adapter 4, and the needle connector is sleeved on the adapter, so that the syringe, the needle connector and the inner tube are connected;

[0015] The piston is nested in the syringe, and the push rod is connected to the piston.

[0016] In some specific embodiments of the present invention, the outer tube of the above-mentioned minimally invasive sampling device has an outer diameter of 1.2 mm, an inner diameter of 0.9 mm, and a length of 30 mm.

[0017] In some specific embodiments of the present invention, the inner tube of the above-mentioned minimally invasive sampling device has an outer diameter of 0.7 mm, an inner diameter of 0.4 mm, and a length of 20 mm.

[0018] In some specific embodiments of the present invention, the side hole of the above-mentioned minimally invasive sampling device is elliptical.

[0019] In some specific embodiments of the present invention, the long axis of the side hole of the above-mentioned minimally invasive sampling device is 1.0 mm, 1.1 mm or 1.2 mm, and the short axis is 0.8 mm, 0.9 mm or 1.0 mm.

[0020] In some specific embodiments of the present invention, the above-mentioned minimally invasive sampling device further comprises a rubber ring 9;

[0021] The rubber ring is nested on the outer surface of the outer tube.

[0022] In some specific embodiments of the present invention, the above-mentioned minimally invasive sampling device further includes a protective cover 10 which is sleeved outside the sampling tube.

[0023] In some specific embodiments of the present invention, the two side holes of the minimally invasive sampling device are opened within 5-10 mm of the tip of the outer tube, and can be 5 mm, 6 mm, 7 mm, 8 mm, 9 mm or 10 mm.

[0024] In some specific embodiments of the present invention, the two side holes of the above-mentioned minimally invasive sampling device open on different sides.

[0025] In some specific embodiments of the present invention, the difference between the distance between one of the two side holes of the above-mentioned minimally invasive sampling device and the tip of the outer tube and the distance between the other side hole and the tip of the outer tube is not less than 1-6 mm, and can be 1 mm, 2 mm, 3 mm, 4 mm, 5 mm or 6 mm.

[0026] In some specific embodiments of the present invention, the side hole diameter of the above-mentioned minimally invasive sampling device is 2-3 mm, which can be 2 mm, 2.1 mm, 2.2 mm, 2.3 mm, 2.4 mm, 2.5 mm, 2.6 mm, 2.7 mm, 2.8 mm, 2.9 mm or 3.0 mm.

[0027] In some specific embodiments of the present invention, the outer tube of the above-mentioned minimally invasive sampling device has scales.

[0028] In some specific embodiments of the present invention, the above-mentioned minimally invasive sampling device is composed of a push rod 1, a syringe 2, a piston 3, an adapter 4, a needle connector 5, an inner tube 6, an outer tube 7, a needle hole 8, a rubber ring 9, and a protective sleeve 10;

[0029] The piston 3 is controlled by the push rod 1, the two are connected and placed in the syringe 2;

[0030] The improved sampling tube is a double-tube structure, consisting of a 20 mm long inner tube 6 and a 30 mm long outer tube 7, which performs a filtering function to prevent the internal organs from being adsorbed and making it impossible to sample;

[0031] The inner tube 6 is connected to the outer tube 7 via a needle connector 5 and fixed to the syringe adapter 4;

[0032] The outer tube 7 is a double-hole side-suction outer tube. Figure 2 As shown, two offset holes 8 are opened on the side, and the surface of the outer tube 7 has unit millimeter markings, with a long line every 5 mm;

[0033] The outer tube 7 is covered with a rubber ring 9 for fixing the required sampling depth. When the device is not in use, the sampling tube is placed in a needle tube protective sleeve 10, and the protective sleeve 10 is connected to the needle connector 5.

[0034] In some specific embodiments of the present invention, the above-mentioned minimally invasive sampling device, the sampling tube specifications are: outer tube: outer diameter * inner diameter * length are 1.2 mm * 0.9 mm * 30 mm respectively;

[0035] Inner tube: outer diameter*inner diameter*length are 0.7 mm*0.4 mm*20 mm respectively.

[0036] The present invention also provides a method for sampling sea cucumber coelomic fluid, which is based on the above-mentioned minimally invasive sampling device for sampling.

[0037] In some specific embodiments of the present invention, the above-mentioned sampling method includes: determining the sampling site of the sea cucumber by ultrasonic detection, using the above-mentioned minimally invasive sampling device to perform ultrasonic puncture, and aspirating and obtaining the sea cucumber body cavity fluid.

[0038] In some specific embodiments of the present invention, the sampling site described in the above sampling method is the central position of the back of the sea cucumber.

[0039] In some specific embodiments of the present invention, the above-mentioned sampling method further includes the step of decontamination and / or disinfection of the sea cucumber after determining the sampling site of the sea cucumber and before performing ultrasonic puncture.

[0040] In some specific embodiments of the present invention, during the puncture by the above-mentioned sampling method, the minimally invasive sampling device penetrates into the sampling site of the sea cucumber at an angle of 40° to 50°.

[0041] In some specific embodiments of the present invention, the above-mentioned sampling method uses an ultrasonic detector to measure the thickness of the body wall before sampling, avoids the visceral area and determines the sampling site to prevent puncturing of the intestines and other internal organs during sampling.

[0042] In some specific embodiments of the present invention, the above-mentioned sampling method comprises four steps: selecting a sampling location for measuring the thickness of the body wall, decontamination and disinfection, sampling the body cavity fluid, and post-processing and observation;

[0043] The selection of the sampling position for measuring the body wall thickness comprises: placing the sea cucumber in seawater, with the seawater covering the sea cucumber by 0.5-1.5 cm, using an ultrasonic detector to measure the body wall thickness of the sea cucumber, finding an area where the body wall is thinner and avoiding the internal organs, adjusting the rubber ring to the corresponding scale of the outer tube according to the measured body wall thickness of the sea cucumber to avoid puncturing the internal organs and increasing damage to the sea cucumber, and the sampling position is preferably the central position of the back of the sea cucumber;

[0044] The decontamination and disinfection treatment includes: preparing filtered sterile seawater to clean the sampling site, repeatedly washing the sampling site with sterile seawater 2 to 3 times to remove surface dirt, impurities and microorganisms;

[0045] The sea cucumber coelomic fluid sampling comprises: using the above-mentioned minimally invasive sampling device, removing the protective cover, adjusting the rubber ring to the required scale according to the body wall thickness measured by the ultrasonic detector, slowly piercing the sampling needle into the central body wall of the back of the sea cucumber at a 45° angle by 3 to 6 mm, and after piercing the body wall, sucking 0.5 to 1 mL of the sea cucumber coelomic fluid, and collecting the collected coelomic fluid into a sampling tube after sampling for subsequent analysis;

[0046] The post-processing and observation include: after the sea cucumber is sampled, the sampling part of the sea cucumber is rinsed again with filtered sterile seawater for 2 to 3 times. The sampled sea cucumber is placed in an independent water tank, 1 / 2 of the water is changed every day, feces is cleaned, the water body is kept clean to prevent the invasion of pathogens, and the survival of the sea cucumber is observed.

[0047] In some specific embodiments of the present invention, in the above-mentioned sampling method, the sampling needle tube slowly penetrates the body wall of the sea cucumber during sampling, and the double holes of the sampling needle are adjusted to be located on both sides of the side facing the body wall.

[0048] In some specific embodiments of the present invention, when the above-mentioned sampling method uses a sampling device to take samples, the push rod is pulled and the internal organs and other tissues suspended in the body cavity fluid are separated by the outer tube of the sampling needle. The body cavity fluid is filtered through the outer tube and enters the outer tube through the small holes on the left and right sides, and is then sucked into the sampling syringe by the negative pressure of the inner tube.

[0049] The minimally invasive and precise sampling method of sea cucumber coelomic fluid of the present invention has the following effects:

[0050] (1) Minimally invasive sampling: This method causes minimal damage to sea cucumbers and does not affect their survival.

[0051] (2) Accurate sampling: Overcoming the blind method does not guarantee that only body cavity fluid is collected and accurate sampling is achieved.

[0052] (3) Easy to operate: The sampling operation is simple and easy. Compared with traditional sampling methods, it effectively shortens the sampling time, is highly efficient, and is suitable for large-scale applications.

[0053] (4) High safety: Sampling tools are properly disinfected to effectively prevent infection.

[0054] (5) Implementation effect: The present invention successfully achieves micro-traumatic sampling of sea cucumbers by using a method of accurately controlling the sampling depth using a modified needle and an ultrasonic detector. After sampling, the sea cucumber is placed in an independent water tank to observe its physiological state. From the second to the third day, the wound of the sea cucumber is basically healed and in good condition, with a survival rate of up to 100%. The coelomocytes of the obtained sea cucumber coelomic fluid are immediately observed under a microscope to determine the sampling accuracy. Experimental verification shows that this method can effectively obtain coelomic fluid samples without affecting the survival of the sea cucumber. This method can achieve minimally invasive and accurate sampling, avoid the damage to the sea cucumber caused by traditional anatomical sampling, reduce the loss of sea cucumbers, and realize repeated research on the same sea cucumber. In addition, this method is simple to operate and easy to master, with high sampling efficiency, providing technical support for biological research and resource protection of sea cucumbers. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] 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 or the description of the prior art are briefly introduced below.

[0056] Figure 1 Schematic diagram of sampling device is shown;

[0057] Figure 2 7 is an enlarged view of the outer tube;

[0058] Figure 3 The schematic diagram of the measurement of sea cucumber body wall thickness and sampling location under ultrasonic detector is shown;

[0059] In the figure, 1-push rod; 2-syringe; 3-piston; 4-adapter; 5-needle connector; 6-inner tube; 7-outer tube; 8-needle hole; 9-rubber ring; 10-protective cover. DETAILED DESCRIPTION

[0060] The present invention discloses a minimally invasive and precise sampling method for sea cucumber coelomic fluid. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0061] The minimally invasive and precise sampling method for sea cucumbers comprises the following steps:

[0062] 1. Place the sea cucumber in seawater with the sea cucumber covered by 1 cm. Use an ultrasonic detector to measure the thickness of the sea cucumber's body wall and find a location where the body wall is thinner and away from the internal organs, such as the center of the sea cucumber's back.

[0063] 2. Install the syringe and take the sample. Connect the 2 ml syringe to the improved sampling needle tube, and slowly insert the needle 3 to 6 mm into the central body wall of the back of the sea cucumber at a 45° angle under the monitoring of the ultrasonic detector to absorb the body cavity fluid of the sea cucumber.

[0064] Before sampling, according to the purpose of sampling, the sampling area, depth and needle trajectory are accurately determined under the monitoring of an ultrasonic detector to avoid the needle piercing the sea cucumber's intestines and other internal organs and achieve accurate sampling.

[0065] As a preferred embodiment of the present invention, the negative pressure sampling of the syringe is used, which is applicable to both large and small sea cucumbers, simplifies the operation, and can be operated by one person.

[0066] As a preferred embodiment of the present invention, real-time monitoring by an ultrasonic detector can determine the needle sampling path, effectively avoid scratching the internal organs, reduce damage to the sea cucumber and achieve accurate sampling.

[0067] As a preferred embodiment of the present invention, the respiratory tree and other internal organs are suspended in the body fluid of the sea cucumber, and the outer tube of the improved needle tube is a double-opening structure on the side, with the opening located at the staggered position of the double needle tubes, as close as possible to the needle head of the outer tube and the two opening positions are staggered; this can prevent the internal organs in the body cavity from being adsorbed when the body cavity fluid is reduced and the negative pressure absorbs the body cavity fluid, blocking the sampling hole and causing failure of successful sampling. The outer tube has a filtering function, and the combination with the inner tube can improve the sampling efficiency and achieve successful sampling.

[0068] It should be understood that the expression "one or more of..." includes each of the items recited after the expression individually and in various different combinations of two or more of the recited items, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited items should be understood to have the same meaning, unless otherwise understood from the context.

[0069] The use of the terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, for example not excluding other unrecited elements or steps, unless otherwise specifically stated or otherwise understood from the context.

[0070] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the application is still operable. In addition, two or more steps or actions can be performed simultaneously.

[0071] The use of any and all examples or exemplary language such as "for example" or "including" herein is intended only to better illustrate the present application and is not intended to limit the scope of the present application. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the present application.

[0072] In addition, the numerical ranges and parameters used to define the present application are approximate values, and the relevant values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in this disclosure are modified by "about". Here, "about" generally refers to the actual value within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0073] Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all common commercial products and can be purchased from the market.

[0074] The present invention will be further described below in conjunction with the embodiments.

[0075] Example 1

[0076] Sea cucumber body cavity fluid collection device Figure 1 As shown, the device includes: a push rod 1; a syringe 2; a piston 3; an adapter 4; a needle connector 5; an inner tube 6; an outer tube 7; a needle hole 8; a rubber ring 9; and a protective sleeve 10. The piston 3 is controlled by the push rod 1, and the two are connected and placed in the syringe 2. The improved sampling tube is a double-tube structure (composed of a 20 mm inner tube 6 and a 30 mm outer tube 7): the inner tube 6 and the outer tube 7 are connected through a needle connector 5 and fixed to the syringe adapter 4. The outer tube 7 is a double-hole side suction type such as Figure 2 As shown, two offset holes 8 are opened on the side, and the surface of the outer tube 7 has unit millimeter markings, with a long line every 5 mm. A rubber ring 9 is sleeved on the needle tube (to mark and fix the sampling depth. After the needle tube reaches the target depth, it is stuck by the rubber ring to prevent the insertion from being too deep and piercing the internal organs). When not in use, the improved needle tube is placed in the needle tube protective cover 10, and the protective cover 10 is connected to the needle connector 5.

[0077] Before sampling, the thickness of the body wall is measured with an ultrasonic detector, and the modified needle is connected to a 2 ml syringe. The needle is soaked with sterile seawater, and the needle is tilted at 45 degrees to the body wall of the sea cucumber. The ultrasonic detector monitors the needle path, and the needle is slowly inserted into the body wall of the sea cucumber, with the depth controlled at 3~6 mm. The coelomic fluid of the sea cucumber is sucked by negative pressure. Immediately after sampling, the coelomic fluid sample is prepared and the coelomic fluid cells are observed under a microscope to determine the sampling accuracy. In the early stage of the experiment, a single-tube side-hole sampling needle with an outer diameter * inner diameter * length of 1.6*1.3*30 mm was used to sample 20 green sea cucumbers. The sampling success rate of a single-tube sampling needle was less than 5%. Under this premise:

[0078] This experimental example evaluates the success rate of sampling sea cucumber coelomic fluid using two types of sampling tubes: outer tube single hole and outer tube double hole with outer diameter*inner tube*length of 1.6*1.3*30mm / 1.2*0.9*20 mm respectively.

[0079] Outer tube single hole / inner tube specifications: 1.6*1.3*30 mm / 1.2*0.9*20 mm, outer tube opening aperture major axis: about 1.5 mm, minor axis: about 1.3 mm; and

[0080] The outer tube double hole / inner tube specifications are: 1.6*1.3*30 mm / 1.2*0.9*20 mm, the outer tube opening aperture major axis: about 1.5 mm, minor axis: about 1.3 mm.

[0081] The coelomic fluid sampling experiment was conducted on sea cucumbers. The two types of needles were used to randomly sample 20 sea cucumbers. The experimental results are shown in Table 1.

[0082] Table 1: Evaluation of the success rate of sampling sea cucumber cavity fluid with two sampling tubes of the same specification, single-hole and double-hole

[0083]

[0084] This embodiment uses different types (single hole in outer tube, double hole in outer tube) of thicker needle tube double tube outer tube / inner tube: 1.6*1.3*30 mm / 1.2*0.9*20 mm to sample the coelomic fluid of sea cucumber using this sampling method. The most suitable sampling needle type is double hole in outer tube / inner tube, and the success rate of single sampling can reach 100%. However, the sampling needle of single hole in outer tube / inner tube is blocked halfway during sampling, which makes it impossible to suck and the coelomic fluid in the syringe is mostly air. The sampling needle is pulled out to adjust the direction of the needle hole and sample again to successfully obtain it, but repeated operations increase the operation time and difficulty, and the stimulation to sea cucumber is also aggravated. Therefore, according to the experimental results, through repeated verification, the most suitable sampling needle type is double tube (double hole in outer tube / inner tube), which has little damage and a sampling success rate of up to 100%.

[0085] Experimental Example 2

[0086] This example evaluates the success rate of coelomic fluid sampling and the survival rate of sea cucumbers using double-tube sampling tubes of different specifications, using the following three specifications of samplers:

[0087] ① Double tubes, double holes in the outer tube: the outer tube outer diameter * inner diameter * length is 1.6 * 1.3 * 30 mm, the inner tube outer diameter * inner diameter * length is 1.2 * 0.9 * 20 mm, the outer tube opening aperture is about 1.5 mm in long axis and about 1.3 mm in short axis;

[0088] ②Double tubes, double holes in the outer tube: the outer tube outer diameter * inner diameter * length is 1.2 * 0.9 * 30 mm, the inner tube outer diameter * inner diameter * length is 0.7 * 0.4 * 20 mm, the outer tube opening aperture major axis is about 1.1 mm, and the minor axis is about 0.9 mm;

[0089] ③ Double tubes, double holes in the outer tube: the outer tube outer diameter * inner diameter * length is 0.9 * 0.6 * 30 mm, the inner tube outer diameter * inner diameter * length is 0.5 * 0.2 * 20 mm, the outer tube opening aperture is about 0.9 mm in major axis and about 0.7 mm in minor axis;

[0090] A sampling experiment was conducted on sea cucumbers. The coelomic fluid of 40 random sea cucumbers was collected using a sampler of each specification. After the coelomic fluid was collected, sections were immediately made to observe the coelomic fluid cells to determine whether the sample was coelomic fluid and to determine the sampling accuracy. The experimental results are shown in Table 2.

[0091] Table 2: Evaluation of different specifications of sampling tubes on the success rate of body cavity fluid sampling and sea cucumber survival rate

[0092]

[0093] Through experimental verification, the sampling efficiency and success rate of the sampling tube with a larger diameter are higher, but it is more likely to cause stress behaviors such as peeling or vomiting, and the wound healing is slower; and because the internal organs are floating in the sea cucumber body cavity fluid, although the smaller the diameter of the sampling tube, the less damage it will cause, it is not easy to take out, and some need to be repeatedly pierced several times to successfully take out. Therefore, the present invention is used to perform negative pressure sampling of the sea cucumber body cavity fluid. For any sea cucumber (maximum 236.73 g, minimum 23.66 g), the optimal specifications are outer tube / inner tube: outer diameter*inner diameter*length 1.2*0.9*30 mm / 0.7*0.4*20 mm, and the sampling success rate, accuracy rate and later survival rate all reach 100%.

[0094] 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 minimally invasive sampling device for sea cucumber coelomic fluid, characterized in that: Includes sampling tube, needle connector, syringe, piston, and push rod; The sampling tube comprises an outer tube and an inner tube, the outer tube is sleeved outside the inner tube, and the same end of the outer tube and the inner tube is connected to the needle connector; The outer tube is a side hole needle having at least two side holes; The syringe is provided with an adapter, and the needle connector is sleeved on the adapter, so that the syringe, the needle connector and the inner tube are connected; The piston is nested in the syringe, and the push rod is connected to the piston.

2. The minimally invasive sampling device according to claim 1, characterized in that: The outer tube has an outer diameter of 1.2 mm, an inner diameter of 0.9 mm, and a length of 30 mm.

3. The minimally invasive sampling device according to claim 1 or 2, characterized in that: The inner tube has an outer diameter of 0.7 mm, an inner diameter of 0.4 mm, and a length of 20 mm.

4. The minimally invasive sampling device according to any one of claims 1 to 3, characterized in that: Also included are rubber rings; The rubber ring is nested on the outer surface of the outer tube.

5. The minimally invasive sampling device according to any one of claims 1 to 4, characterized in that: It also includes a protective cover which is sleeved outside the sampling tube.

6. The minimally invasive sampling device according to any one of claims 1 to 5, characterized in that: The outer tube has graduations.

7. A method for sampling sea cucumber coelomic fluid, characterized in that: Sampling is performed based on the minimally invasive sampling device according to any one of claims 1 to 6.

8. The sampling method according to claim 7, characterized in that: include: The sampling site of the sea cucumber is determined by ultrasonic detection, and the minimally invasive sampling device described in any one of claims 1 to 6 is used for puncture to absorb and obtain the sea cucumber body cavity fluid.

9. The sampling method according to claim 8, characterized in that: After determining the sampling site of the sea cucumber and before puncturing, the process also includes steps for decontamination and / or disinfection of the sea cucumber.

10. The sampling method according to claim 8 or 9, characterized in that: During puncture, the minimally invasive sampling device pierces the sampling site of the sea cucumber at an angle of 40° to 50°.

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