CN109406215B - Intelligent acquisition device and acquisition method for water body environment DNA - Google Patents
Intelligent acquisition device and acquisition method for water body environment DNA Download PDFInfo
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- G01N1/00—Sampling; Preparing specimens for investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/10—Devices for withdrawing samples in the liquid or fluent state
- G01N1/14—Suction devices, e.g. pumps; Ejector devices
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Abstract
The utility model discloses an intelligent acquisition device and an intelligent acquisition method for water body environment DNA, and belongs to the technical field of environment DNA detection. The intelligent water body environment DNA collecting device and the collecting method provided by the utility model integrate intelligent control, sample collection, residual night emptying and pipeline blockage and pressure relief protection functions, can collect biological samples of water body, can collect water samples, can effectively improve the efficiency of the biological sample sampling process, ensure the accuracy of water filtering quantity calculation, ensure the timeliness of the samples, and enable the environment of detecting the water samples to be more similar to the natural environment; has the advantages of difficult blockage, simple structure, reasonable design and easy manufacture.
Description
Technical Field
The utility model belongs to the technical field of environmental DNA detection, and particularly relates to an intelligent acquisition device and an intelligent acquisition method for water body environmental DNA.
Background
Species distribution and species abundance are major problems in species diversity research of freshwater ecosystems, and play a vital role in implementing biodiversity protection measures. The DNA obtained from the living environment of the species reflects the information of the distribution, abundance and the like of the species, and is a non-invasive and rapid species detection means. With the rapid development of high-throughput sequencing technology, species diversity research methods based on environmental DNA and molecular biology technology (eDNA macro barcode technology) are increasingly applied to water ecological health evaluation, and the recognition level of species diversity of an ecological system is remarkably improved. Researchers such as Ficetda (Species detection using environmental DNA from water samples [ J ] Biol Lett,2008,4 (4): 432-435) have successfully used environmental DNA detection methods to detect the presence of invasive species bullfrog in natural ponds.
It is well known that the error of an analytical process can be gradually reduced by technological advances, but the sampling technique is not sufficiently long over a long period of time, so that the total error of the analytical measurement end result is often from the sampling process. Unlike the sampling mode of acquiring biological individual samples in the traditional biodiversity investigation, the DNA macro-bar code technology needs to acquire the total environmental DNA in the receiving water body rapidly, so that the representativeness, the accuracy and the integrity of the acquired samples become the primary problems of water environment DNA monitoring to acquire correct and reliable analysis results. Proper selection of sampling methods and apparatus, improved sampling techniques, are extremely important for improving analytical monitoring quality. Conversely, if sampling is wrong, the subsequent analysis process will lose meaning and even cause huge waste.
The traditional sample collection method is mainly based on manual collection, and the subsequent dehydration treatment is carried out in a laboratory to obtain an environment DNA collection sample, such as Chinese patent application, and the issued bulletin number: CN203811429U, authorized bulletin day: 2014.09.3A net for collecting qualitative sample of plankton in rapid water body comprises a bag-shaped net body, wherein a valve is arranged at the tail end of the front end of the net body, a hollow pipe hoop is arranged in the middle of the net body, the net body is divided into a funnel-shaped net body and a net belly positioned behind the net body and communicated with the net body by the pipe hoop, and the net can be widely used in the field of plankton sample collection. However, this method has the following significant drawbacks: (1) manual operation, the efficiency is low: the method is greatly constrained by coastal water bodies, ships and the like, the sampling range is limited, and the sample representativeness is poor; and the labor and material resources are wasted, and the working efficiency is low; (2) the sampling position is difficult to accurately control: under the influence of water flow, a plankton net is easy to float away from a sampling point during sampling, and a plurality of factors such as a ship influence, so that the ship cannot be sampled in certain water areas; and (3) the calculation error of the filtered water amount is larger: because of the change of the sampling depth of the plankton net, the estimated water filtering quantity according to the release length, the sampling angle and the sampling time of the steel wire rope has larger error, and the plankton density calculated by the water filtering quantity is not consistent with the actual situation; (4) In the period from collection to analysis and measurement, the water environment sample is extremely easy to change, and the change of the collected sample can be caused due to the influence of the metabolic activity, the chemical action and the physical action of organisms in the water body, so that the change, even degradation, of the environmental DNA is caused, the detection rate of the species is further influenced, and meanwhile, the detection results such as the distribution of the species are also influenced. And these changes are rapid, so that the necessary protective measures must be taken to preserve the sample. Although the eDNA macro bar code technology is used as an emerging environment monitoring means to greatly improve the efficiency and accuracy of environment monitoring, the traditional biological sampling method does not meet the requirement of the novel monitoring technology, and the large-scale popularization and application of the method in an environment-friendly system are limited. Therefore, when sampling the water environment, the quality of the sampling is guaranteed and controlled very necessarily. Such as chinese utility model patent, filing date: 2016.05.25, application number: 201620486713.8A water body environment DNA sampling device comprises a protective cover, a pull rod, a water pump and a water pumping pipeline connected with the pull rod, wherein filter paper is arranged at the other end of the water pumping pipeline, the filter paper is fixed through a fixed ring, the fixed ring also comprises a plurality of fan blades for covering the filter paper, the fan blades are matched to form a circle, one corner of each fan blade is connected with a fixed ring shaft, and the edges of the fan blades are connected to the protective cover through connecting pipes; the utility model can complete the on-site collection work of the water body environment DNA without the subsequent heavy filtering operation, saves time and labor, but still has the advantages of incapability of quantitatively collecting, large water filtering quantity error, and particularly, the equipment is easy to block in the use process, the normal use is influenced by light weight, and the pipeline is damaged by heavy weight
In addition, it is worth noting that species enrichment is based on ambient DNA concentration, which is affected by the release and degradation process of DNA, as well as ambient DNA extraction methods and water sample volumes. The eDNA macro barcode technology is mainly based on obtaining plankton (including plant, animal, microorganism and organisms with different sizes) in a receiving water body, suspended particulate matters in a water sample and the like to detect environmental DNA, so that if the DNA in the corresponding water body environment is truly reflected, the real effectiveness of the collected sample needs to be ensured.
To minimize the above-mentioned changes, certain protection measures are generally taken on the sample, such as measures of refrigeration and freezing or adding stabilizers; then, the transport time is shortened as much as possible, and the analysis and measurement are performed as soon as possible. For example, chinese patent application publication No. 2018.07.17 and publication No. CN 105636937B disclose a stabilizer for preserving biological samples, wherein the stabilizer is mixed with biological samples according to a certain proportion when in use, and the integrity and stability of biological samples can be maintained for a long time under non-freezing condition by using the stabilizer; there is still a risk that the added stabilizer interferes with the visual inspection of the monitoring item.
Therefore, the development of the environmental DNA sample collection equipment improves the existing sampling technology, improves the sampling efficiency, reduces the influencing and interfering factors in the sampling process, reduces the influencing factors of the environmental DNA extraction method and the water sample volume on the environmental DNA, and is important for improving the analysis and monitoring quality of the species richness of the water body environment.
Disclosure of Invention
1. Problems to be solved
Aiming at the problems that the efficiency of the existing water body environment biological sample sampling process is low, the calculation error of the filtered water quantity is large, the biological sample is easy to change after the sampling is carried out, and the existing sampling equipment is easy to block and cause pipeline damage, the utility model provides the intelligent water body environment DNA collecting device and the collecting method.
2. Technical proposal
In order to solve the problems, the technical scheme adopted by the utility model is as follows:
the intelligent water body environment DNA acquisition device comprises an intelligent control mechanism and a sampling main body mechanism, wherein the sampling main body mechanism comprises a sample acquisition unit, a pressure relief unit and a water emptying unit;
the sample collection unit comprises a water pump, a collector and a flowmeter, wherein one end of the water pump is connected with an external water body, and the other end of the water pump is connected with a water inlet of the collector; the flowmeter is connected with the water outlet of the collector;
the water emptying unit comprises an air inlet pipeline and a vacuum pump, one end of the air inlet pipeline is connected with the water inlet of the collector, and the other end of the air inlet pipeline is communicated with the external atmosphere; one end of the vacuum pump is connected with the flowmeter, and the other end of the vacuum pump is connected with the external environment;
the pressure relief unit comprises a pressure relief valve, and the pressure relief valve is arranged between the water pump and the collector and is communicated with the external atmosphere;
the intelligent control mechanism comprises a main control board and a GPS positioning block connected with the main control board, wherein the main control board is used for controlling the opening or closing of the acquisition unit and the water draining unit and receiving positioning information of the GPS positioning block.
Preferably, the sampling main body mechanism further comprises a three-way valve, and a first inlet of the three-way valve is connected with an external water body; the outlet of the three-way valve is connected with a water pump; the second inlet of the three-way valve is connected with an air inlet pipe of the water emptying unit.
Preferably, the sample collection unit further comprises a coarse filter, an electromagnetic valve II and a one-way valve; the coarse filter is arranged on a first inlet pipeline of the three-way valve, and an electromagnetic valve II is arranged on a pipeline between the coarse filter and the first inlet of the three-way valve; the one-way valve and the vacuum pump are connected in parallel and connected in the pipeline.
Preferably, the water draining unit further comprises a solenoid valve I, wherein the solenoid valve I is positioned on the air inlet pipeline.
Preferably, the collector is an enrichment collector for biological enrichment of the water sample, and a filtering membrane is arranged in the enrichment collector; the collector can also be a collecting bottle for collecting water samples.
Preferably, the enrichment collector comprises an upper seat body, a middle seat body and a lower seat body; the upper seat body is in clamping connection with the middle seat body to form an upper cavity; the middle seat body and the lower seat body are connected in a clamping way to form a lower chamber, and a filtering membrane is arranged between the upper chamber and the lower chamber.
The number of the middle seats can be one or more, and the adjacent middle seats are in clamping connection and are provided with filtering membranes with different apertures; the filtering pore diameter of the filtering membrane is 0.2-200 microns.
When a plurality of filtering membranes exist, the pore diameters of the filtering membranes are gradually decreased from large to small in the water inlet direction.
Preferably, the intelligent control mechanism further comprises a power supply system, a display screen and a hand control board which are respectively connected with the main control board; the display screen displays real-time operation parameters of the device; the manual control board is used for carrying out parameter presetting on the main control board;
the intelligent control mechanism also comprises a wireless control board and a remote controller which is in wireless connection with the wireless control board, wherein the wireless control board is simultaneously connected with the main control board and the GPS positioning block, and the remote controller inputs control instructions to the main control board through the wireless control board.
The main control board is connected with the power supply system, the display screen and the hand control board; the set parameters of the manual control plate can be read in real time, and the calculation is converted into the action and the running state of a control system such as a pump, a valve and the like respectively; and simultaneously, the running state is reversely displayed through the display unit.
Preferably, the intelligent water body environment DNA collecting device can be switched through a plurality of electromagnetic pipelines to implement multi-pipeline water sample biological enrichment or water sample collection. .
An intelligent acquisition method for water body environment DNA comprises the following steps:
A. constructing the water body environment DNA acquisition device;
B. parameter presetting: presetting sampling parameters of the main control board during device operation by using the manual control board;
C. sample collection: after the acquisition device is placed at an acquisition place, the system is started, the main control board sends out an instruction according to instruction parameters, the sampling unit is controlled to work, the electromagnetic valve II and the water pump are started, and a water sample enters the collector to sample under the driving of the water pump; at this time, the water emptying unit and the pressure relief unit are in a non-working state; enriching the biological sample of the water body in the water sample into an enrichment collector; the redundant water body flows through the flowmeter, the device is discharged through the one-way valve, the flowmeter accumulates the water passing amount in the reading device in real time and transmits signals to the main control board, and when the water passing amount is consistent with the preset flow parameter, the work of the acquisition unit is finished; the main control board controls the water emptying unit to work, the water enriched with the sample is emptied, and after the water emptying is completed, the water biological sample collection is completed;
D. and (3) water emptying: after the work of the collecting unit is finished, the main control board controls the water emptying unit to work, the vacuum pump is started to empty residual liquid in the enrichment collector, after the water is emptied, one working process is finished, and the next sampling can be performed by replacing the collector;
E. decompression: if the blockage phenomenon occurs in the sample collection process, the pipeline pressure can be excessively large, and at the moment, if the pipeline pressure is larger than the critical pressure value of the pressure relief valve, the pressure relief valve is automatically opened to form a liquid circulation pipeline, the pipeline pressure is regulated, and the device is effectively protected.
The working principle of the device of the utility model is as follows:
parameter presetting: utilize manual control board to water total flow parameter V 1 Setting parameters such as water emptying operation time and the like and transmitting the parameters to a main control module;
sample collection: after the acquisition device reaches an acquisition site, a starting command is output by utilizing a remote control device, the starting command is sent by a wireless signal transceiver, a wireless communication module on a wireless control board receives a signal and feeds the signal back to a main control board, the main control board controls a solenoid valve II and a water pump to be started, a water body enters a pipeline through a coarse filter under the driving of the water pump, the water body enters a collector through the solenoid valve II, a three-way valve and the water pump, biological samples in the water body are enriched on a filtering membrane, and redundant residual liquid flows through a flowmeter and is discharged out of the device through a one-way valve; in the process, the flowmeter accumulates and reads the water flow t in the device in real time 2 And transmits the signal to the main control board when the water supply amount V is over 2 And the total flow parameter V 1 When the signals are consistent, the main control board outputs signals to control and close the electromagnetic valve II and the water pump;
and (3) water emptying: then, the main control board outputs a signal, controls and opens the electromagnetic valve I, starts the vacuum pump to perform delayed operation and evacuation, and under the negative pressure adsorption action of the vacuum pump, external atmosphere is sucked through the air inlet pipe and sequentially passes through the electromagnetic valve I, the water pump, the collector, the flowmeter and the vacuum pump, and finally the device is discharged, so that residual liquid in the collector is evacuated, and after the water evacuation operation time is reached, the system automatically stops; at this time, the sampling work process is finished, and the next sampling is carried out by replacing the collector;
decompression: if a blockage phenomenon occurs in the sample collection process and a pipeline is not smooth, the pipeline pressure is overlarge, at the moment, (if the pipeline pressure is larger than a critical pressure value of a pressure relief valve), the pressure relief valve is automatically opened to form a circulating pipeline for discharging liquid, the pipeline pressure is regulated, and an effective protection device is provided;
in the running process of the device, the main control board reads the set parameters and converts the set parameters into signals, controls the actions and running states of all parts of the pipeline in real time, compares and calculates the read parameters with the set values, and reversely displays the parameters through the display device, so that the device is an intelligent control core processing unit; the power supply system is used for providing electric energy, the manual control board is used for presetting parameters of the main control board, the wireless control board is used for transmitting signals between the main control board and the remote controller, and real-time operation parameters of equipment are displayed on the display screen; the GPS positioning plate is used for positioning in real time and feeding back to the main control plate and the remote controller, and displaying on the display screen and the display screen of the remote controller.
3. Advantageous effects
Compared with the prior art, the utility model has the beneficial effects that:
(1) The intelligent water body environment DNA collecting device and the collecting method provided by the utility model integrate intelligent control, sample collection, residual night emptying and pipeline blockage and pressure relief protection functions, and have the following effects:
(1) the intelligent program control of the singlechip can automatically operate according to preset parameters, accurate sampling is realized, a GPS system is arranged, the accurate positioning sampling automation degree is high, personnel are prevented from going to a sampling area by means of a motor ship, the sampling efficiency is high, and the cost of manpower and material resources is saved;
(2) in the process of sample collection, the flowmeter is arranged behind the collector, can accumulate and read the water yield in real time, and automatically stops sampling after reaching the preset water yield parameter, so that the accurate control of the water yield is realized, the problem of larger water yield filtering error in the prior art is overcome, the accurate quantitative enrichment and sampling function sample of the water environment DNA are truly realized, the calculation precision of plankton density is improved, the sampled sample is more close to the actual situation, and the accuracy and representativeness of the sample are improved;
(3) after the sample collection is completed, residual liquid in the enrichment collector can be emptied to the maximum extent through the water emptying unit, so that the time from collection to analysis and measurement is avoided, and under the condition that the residual water exists, the change of environmental DNA caused by the influence of metabolic activity, chemical action and physical action of organisms in a water body is even degradation due to the change of environmental conditions, thereby influencing the detection rate of species, ensuring the timeliness of the sample to the maximum extent and enabling the environment of a detected water sample to be more similar to the natural environment;
(4) the water body environment DNA acquisition device provided by the utility model truly realizes on-site in-situ enrichment through the combination of positive pressure filtration and negative pressure adsorption;
(5) in the prior art for collecting biological samples of water, the device is easy to be blocked because the collecting and filtering device is provided with a limit of a filtering aperture, and equipment is damaged or even destroyed if timely treatment cannot be performed; the water environment DNA acquisition device provided by the utility model is provided with the pressure relief function, if the blockage phenomenon occurs in the sample acquisition process, the pipeline pressure is larger than the critical pressure value of the pressure relief valve, the pressure relief valve is automatically opened to form a liquid circulation pipeline, the pipeline pressure is regulated, and the device is effectively protected.
(2) According to the intelligent water body environment DNA collecting device and the collecting method, the enrichment collector is provided with the multi-layer filtering membrane, the biological water body sample is directly fixed on the filtering membrane, the working section is completely in a vacuum state, a large amount of water sample is not required to be collected and brought back to a laboratory for later water body biological sample separation, pollution possibly brought by the later water body sample separation operation process is avoided, the water body biological sample is in an original collected state as much as possible, and the reliability of sample treatment is greatly increased;
the enrichment collector is additionally provided with a plurality of groups of multi-level filtering membranes, the pore diameters of the filtering membranes are gradually decreased from large to small according to the direction from water inlet to water outlet, the filtering of different specification granularity grades can be realized, the water sample can pass through the collector to the greatest extent, the working efficiency is improved, and the blockage is avoided.
(3) The intelligent acquisition device and the acquisition method for the water body environment DNA, provided by the utility model, are provided with wireless remote control equipment, can be used for fixed or long-distance remote control sampling, and have the advantages of high automation degree, simplicity, easiness in use, high sampling efficiency and good reliability.
Drawings
FIG. 1 is a front elevational view of the overall structure of the apparatus of the present utility model;
FIG. 2 is a schematic diagram of a remote control of the device of the present utility model;
FIG. 3 is a schematic diagram of the overall structure of the enrichment collector;
FIG. 4 is a schematic view of the middle housing structure of the enrichment collector;
FIG. 5 is a schematic view of a collection bottle of the sample collector;
in the figure: 1. a case; 2. a solenoid valve I; 3. a solenoid valve II; 4. a collector; 410. an upper base; 420. a middle seat body; 421. a boss; 422. a water passing hole; 423. a groove; 430. a lower base; 440. a filtering membrane; 450. a water inlet; 460. a water outlet; 5. a flow meter; 6. a vacuum pump; 7. a water inlet pipe; 8. a coarse filter; 9. a one-way valve; 10. a main control board; 11. a manual control plate; 111. a manual control button; 12. a GPS positioning block; 13. a wireless control board; 14. an antenna; 15. a power supply system; 16. a display screen; 17. a remote control device; 171. a wireless signal transceiver; 172. a remote control display screen; 173. a control button; 18. a water pump; 19. a pressure release valve; 20. and a three-way valve.
Detailed Description
The utility model is further described below in connection with specific embodiments.
It will be understood that when an element is referred to as being "mounted" on another element, it can be directly on the other element or intervening elements may also be present; when an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present; the terms "vertical," "horizontal," "left," "right," and the like are used herein for illustrative purposes only.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this utility model belongs; the terminology used herein in the description of the utility model is for the purpose of describing particular embodiments only and is not intended to be limiting of the utility model; the term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Example 1
As shown in FIG. 1, the intelligent water body environment DNA acquisition device comprises a box body 1, an intelligent control mechanism and a sampling main body mechanism, wherein the intelligent control mechanism and the sampling main body mechanism are positioned in the box body 1; the sampling main body mechanism comprises a sample collection unit, a pressure relief unit, a water evacuation unit and a three-way valve 20;
the intelligent control mechanism in the embodiment comprises a main control board 10, a GPS positioning block 12 connected with the main control board 10, a power supply system 15, a display screen 16 and a hand control board 11 connected with the main control board 10, and a remote controller 17 connected with a wireless control board 13 in a wireless way; the main control board 10 is used for controlling the opening or closing of the acquisition unit and the water draining unit and receiving positioning information of the GPS positioning block 12. The hand control panel 11 is used for presetting parameters of the main control panel 10 through a hand control button 111, and the display screen 16 displays operation parameters of the main control panel 10; the hand control board 11 is used for carrying out parameter presetting to the main control board 10, the remote controller 17 inputs control instructions to the main control board 10 through the wireless control board 13 and the remote controller 17 is used for realizing the starting and stopping of the remote real-time remote control equipment.
Further, the remote control device in this embodiment is provided with a wireless signal transceiver 171, a remote control display screen 172 and a control button 173, and the control button 173 is used to input a command, the command is transmitted to the wireless control board 13 by the wireless signal transceiver 171, and the remote control display screen 172 is synchronized with the display screen 16 to display the current operation parameters of the device.
The sample collection unit in the embodiment comprises a water pump 18, a collector 4, a flowmeter 5, a coarse filter 8, an electromagnetic valve II 3 (the electromagnetic valve II 3 in the embodiment is a normally closed electromagnetic valve, the model is De-Lixi 2W series voltage is 24V), and a one-way valve 9, wherein the coarse filter 8 is arranged on a first inlet pipeline of the three-way valve 20, and the electromagnetic valve II 3 is arranged on a connecting pipeline between the coarse filter 8 and the first inlet of the three-way valve 20; the outlet of the three-way valve 20 is connected with one end of the water pump 18; the other end of the water pump 18 is connected with the water inlet 450 of the collector 4, and one end of the flowmeter 5 is connected with the water outlet 460 of the collector 4; the main control board 10 is connected with the water pump 18 and the electromagnetic valve II 3 and is used for controlling the opening and closing of the water pump 18 and the electromagnetic valve II 3; when the remote controller 17 inputs and starts the main control board 10 through the wireless control board 13After the instruction, the main control board 10 controls the water pump 18 and the electromagnetic valve II 3 to be opened, under the driving of the water pump 18, water enters a pipeline through the coarse filter 8, enters the collector 4 through the electromagnetic valve II 3, the three-way valve 20 and the water pump 18, the biological sample in the water is enriched on the filtering membrane 440, and the redundant residual liquid flows through the flowmeter and is discharged out of the device through the one-way valve 9; in the process, the flowmeter 5 accumulates and reads the water flow t in the device in real time 2 And transmits the signal to the main control board 10, when the water is over-flowing V 2 And the total flow parameter V 1 When the signals are consistent, the main control board outputs signals to control and close the electromagnetic valve II 3 and the water pump 18.
The water emptying unit in the embodiment comprises an air inlet pipeline, an electromagnetic valve I2 (the electromagnetic valve I2 in the embodiment is a normally closed electromagnetic valve, the model is De-Lixi 2W series voltage is 24V) and a vacuum pump 6, wherein one end of the air inlet pipeline is communicated with the outside atmosphere, and the other end of the air inlet pipeline is connected with a water inlet 450 of the collector 4 through a second inlet of the three-way valve 20; one end of the vacuum pump 6 is connected with the flowmeter 5, and the other end is connected with the external environment; the vacuum pump 6 and the one-way valve 9 are connected in parallel and connected in a pipeline; the main control board 10 is connected with the vacuum pump 6 and the electromagnetic valve I2 and used for controlling the opening and closing of the vacuum pump 6 and the electromagnetic valve I2; after the sampling is finished, the main control board 10 outputs a signal, controls and opens the electromagnetic valve I2, starts the vacuum pump 6, carries out delayed operation and evacuation, and under the negative pressure adsorption action of the vacuum pump 6, external atmosphere is sucked through the air inlet pipe and sequentially passes through the electromagnetic valve I2, the water pump 18, the collector 4, the flowmeter 5 and the vacuum pump 6, and finally the device is discharged, so that residual liquid in the collector 4 is evacuated, and after the water evacuation operation time is reached, the system is automatically stopped; at this time, the sample collection process is finished, and the next sampling is performed by replacing the collector 4;
the pressure relief unit in this embodiment includes a pressure relief valve 19, the pressure relief valve 19 being provided between the water pump 18 and the collector 4 and communicating with the outside atmosphere; normally, the pressure release valve 19 is in a normally closed state, if a blockage phenomenon occurs in the sample collection process, the pipeline is not smooth, the pipeline pressure is overlarge, and at the moment, if the pipeline pressure is larger than the critical pressure value of the pressure release valve 19), the pressure release valve 19 is automatically opened to form a circulation pipeline for discharging liquid, the pipeline pressure is regulated, and an effective protection device is provided;
the collector in the embodiment can be an enrichment collector for enriching biological samples of the water sample, and mainly collects plankton, suspended particles in the water sample and the like; the enrichment collector comprises an upper seat 410, a middle seat 420 and a lower seat 430; the water inlet 450 is positioned at the top end of the upper seat body 410, the lower part of the upper seat body 410 is a cavity, and the outer side wall of the cavity is provided with a boss; the middle seat 420 is divided into an upper cavity and a lower cavity, the inner wall of the upper cavity is provided with a groove 423, the outer side wall of the lower cavity is provided with a boss 421, and a water passing hole 422 is arranged between the upper cavity and the lower cavity; the bottom of the lower seat body is provided with a water outlet 460, and the inner side wall of the cavity body is provided with a boss; the upper base 410 is connected with the concave groove 423 of the upper cavity inner wall of the middle base 420 through the boss of the outer side wall of the cavity to form an upper cavity, and the middle base 420 is connected with the concave groove of the inner wall of the lower base 430 through the boss 421 of the outer side wall of the lower cavity to form a lower cavity; a sealing ring and a filtering membrane 440 are installed between the middle seat 420 and the upper seat 410 and between the middle seat 420 and the lower seat 430; the number of the middle seat bodies 420 can be increased or decreased as required, and is set to be one or more, the adjacent middle seat bodies 420 are in clamping connection, and a sealing ring and filtering membranes 440 with different apertures are arranged in the middle; the filter pore diameter of the filter membrane 440 is 0.2-200 micrometers; the pore diameter of the filtering membrane 440 decreases from large to small in the arrangement direction from the water inlet 450 to the water outlet 460.
After the water body flows through the enrichment collector, the biological sample in the water body is enriched on the filtering membrane 440, after enrichment is finished, the collector 4 is opened, the filtering membrane 440 is clamped by sterile forceps, and the water body is placed into a sterile centrifuge tube (the volume of the centrifuge tube can be 2-10 milliliters) which is prepared in advance, and the water body is stored (freezing storage and room temperature storage can be selected). It should be noted that the collector 4 in this embodiment may be a collection bottle for collecting water samples for extracting nucleic acids, including DNA, RNA and analysis of environmental pollutants.
When the device is matched with a remote control device such as an unmanned ship or an unmanned plane, the function of the device is particularly remarkable, and the device can be positioned in real time through the GPS positioning block 12. After the device is remotely controlled to the collection point through the carrier (unmanned ship unmanned plane) in some specific environments, the running state and specific position of the device are difficult to be seen by remote people. The device can confirm the position in real time through the set GPS positioning function, simultaneously can control the running and display state of the device in real time through the wireless module, and realizes the starting and stopping of long-distance real-time remote control equipment through the remote controller 17.
The foregoing description is not intended to limit the utility model to the particular embodiments disclosed, but on the contrary, the intention is to cover all modifications, alternatives, and equivalents falling within the spirit and scope of the utility model.
Example 2
The intelligent sampling method for the water body environment DNA comprises the following specific steps:
A. constructing a water body environment DNA acquisition device according to the claims 1-8;
B. parameter presetting: the method comprises the steps that sampling parameters are preset when the main control board 10 is operated by the aid of the manual control board 11;
C. sample collection: after the acquisition device is placed at an acquisition place, the system is started, the main control board 10 sends out an instruction according to instruction parameters, the sampling unit is controlled to work, the electromagnetic valve II 3 and the water pump 18 are started, and a water sample enters the collector 4 to sample under the driving of the water pump 18; at this time, the water emptying unit and the pressure relief unit are in a non-working state; enriching the biological sample of the water body in the water sample into an enrichment collector; the redundant water body flows through the flowmeter 5, the device is discharged through the one-way valve 9, the flowmeter 5 accumulates the water passing amount in the reading device in real time and transmits signals to the main control board 10, and when the water passing amount is consistent with the preset flow parameter, the collecting unit finishes working; the main control board 10 controls the water emptying unit to work, the water enriched with the sample is emptied, and after the water is emptied, the water biological sample collection is completed;
D. and (3) water emptying: after the work of the collection unit is finished, the main control board 10 controls the water emptying unit to work, the vacuum pump 6 is started to empty the residual liquid in the enrichment collector, after the water is emptied, one working process is finished, and the next sampling can be performed by replacing the collector 4;
E. decompression: if a blockage phenomenon occurs during the sample collection process, the pipeline pressure will be too high, and at this time, if the pipeline pressure is greater than the critical pressure value of the pressure release valve 19, the pressure release valve 19 will be automatically opened to form a liquid circulation pipeline, and the pipeline pressure is adjusted to provide an effective protection device.
The sampling method in the embodiment has the advantages of accurate sampling, high automation degree, high sampling efficiency and labor and material cost saving. In the collection process, the water yield can be read in real time, and after the water yield reaches the preset water yield parameter, the sampling is automatically stopped, so that the accurate control of the water yield is realized, the problem of larger water yield filtering error in the prior art is overcome, the accurate quantitative enrichment and sampling of the DNA of the water environment are truly realized, the calculation precision of the plankton density is improved, the sampled product is more close to the actual situation, and the accuracy and the representativeness of the sample are improved;
after the sample collection is completed, residual liquid can be emptied to the maximum extent through the water emptying function, the situation that under the condition that the residual water exists from collection to analysis and measurement is avoided, due to the change of environmental conditions, the change of environmental DNA caused by the influence of metabolic activity, chemical action and physical action of organisms in a water body is even degradation, the detection rate of species is further influenced, the timeliness of the sample is ensured to the maximum extent, and the environment for detecting the water sample is more similar to the natural environment.
The foregoing examples merely illustrate certain embodiments of the utility model and are described in more detail and are not to be construed as limiting the scope of the utility model; it should be noted that it is possible for a person skilled in the art to make several variants and modifications without departing from the concept of the utility model, all of which fall within the scope of protection of the utility model; accordingly, the scope of protection of the present utility model is to be determined by the appended claims.
Claims (9)
1. An intelligent water body environment DNA acquisition device which is characterized in that: the intelligent sampling device comprises an intelligent control mechanism and a sampling main body mechanism, wherein the sampling main body mechanism comprises a sample acquisition unit, a pressure relief unit and a water emptying unit;
the sample collection unit comprises a water pump (18), a collector (4) and a flowmeter (5), wherein one end of the water pump (18) is connected with an external water body, and the other end of the water pump is connected with a water inlet (450) of the collector (4); the flowmeter (5) is connected with a water outlet (460) of the collector (4);
the water emptying unit comprises an air inlet pipeline and a vacuum pump (6), one end of the air inlet pipeline is connected with a water inlet (450) of the collector (4), and the other end of the air inlet pipeline is communicated with the outside atmosphere; one end of the vacuum pump (6) is connected with the flowmeter (5), and the other end is connected with the external environment;
the pressure relief unit comprises a pressure relief valve (19), and the pressure relief valve (19) is arranged between the water pump (18) and the collector (4) and is communicated with the external atmosphere;
the intelligent control mechanism comprises a main control board (10) and a GPS positioning block (12) connected with the main control board (10), wherein the main control board (10) is used for controlling the opening or closing of the acquisition unit and the water emptying unit and receiving positioning information of the GPS positioning block (12);
the sampling main body mechanism further comprises a three-way valve (20), and a first inlet of the three-way valve (20) is connected with an external water body; the outlet of the three-way valve (20) is connected with a water pump (18); the second inlet of the three-way valve (20) is connected with an air inlet pipe of the water emptying unit.
2. The intelligent water environment DNA collection device according to claim 1, wherein: the sample collecting unit also comprises a coarse filter (8), an electromagnetic valve II (3) and a one-way valve (9); the coarse filter (8) is arranged on a first inlet pipeline of the three-way valve (20), and an electromagnetic valve II (3) is arranged on a pipeline between the coarse filter (8) and the first inlet of the three-way valve (20); the one-way valve (9) and the vacuum pump (6) are connected in parallel and connected in a pipeline.
3. The intelligent water environment DNA collection device according to claim 1, wherein: the water emptying unit also comprises an electromagnetic valve I (2), and the electromagnetic valve I (2) is positioned on the air inlet pipeline.
4. The intelligent water environment DNA collection device according to claim 1, wherein: the collector (4) is an enrichment collector and is used for biological enrichment of the water sample, and a filtering membrane (440) is arranged in the enrichment collector.
5. The intelligent water environment DNA collection device according to claim 1, wherein: the collector (4) is a collecting bottle and is used for collecting water samples.
6. The intelligent water environment DNA collection device according to claim 4, wherein: the enrichment collector comprises an upper seat body (410), a middle seat body (420) and a lower seat body (430); the upper base body (410) and the middle base body (420) are connected in a clamping mode to form an upper cavity, the middle base body (420) and the lower base body (430) are connected in a clamping mode to form a lower cavity, and a filtering membrane (440) is arranged between the upper cavity and the lower cavity.
7. The intelligent water environment DNA collection device according to claim 6, wherein: the number of the middle seats (420) of the enrichment collector is one or more, and the adjacent middle seats (420) are connected in a clamping way and are provided with filtering membranes (440) with different apertures; the filtering pore diameter of the filtering membrane (440) is 0.2-200 microns;
when a plurality of filtering membranes (440) exist, the pore diameters of the filtering membranes (440) are gradually decreased from large to small in the water inlet direction.
8. The intelligent collection device for DNA in a water environment according to any one of claims 1 to 7, wherein: the intelligent control mechanism also comprises a power supply system (15), a display screen (16) and a hand control board (11), which are respectively connected with the main control board (10); the display screen (16) displays real-time operation parameters of the device; the manual control board (11) is used for carrying out parameter presetting on the main control board (10);
the intelligent control mechanism also comprises a wireless control board (13) and a remote controller (17) which is in wireless connection with the wireless control board (13), wherein the wireless control board (13) is simultaneously connected with the main control board (10) and the GPS positioning block (12), and the remote controller (17) inputs control instructions to the main control board (10) through the wireless control board (13).
9. An intelligent acquisition method for water body environment DNA comprises the following steps:
A. constructing an intelligent water body environment DNA acquisition device according to any one of claims 1-8;
B. parameter presetting: the method comprises the steps that sampling parameters are preset when a main control board (10) is operated by a manual control board (11);
C. sample collection: after the acquisition device is placed at an acquisition place, the system is started, the main control board (10) sends out an instruction according to instruction parameters, the sampling unit is controlled to work, the electromagnetic valve II (3) and the water pump (18) are started, and a water sample enters the collector (4) under the driving of the water pump (18) to sample; at this time, the water emptying unit and the pressure relief unit are in a non-working state; enriching the biological sample of the water body in the water sample into an enrichment collector; the redundant water body flows through the flowmeter (5), the device is discharged through the one-way valve (9), the flowmeter (5) accumulates the water passing amount in the reading device in real time, signals are transmitted to the main control board (10), and when the water passing amount is consistent with the preset flow parameter, the collecting unit finishes working; the main control board (10) controls the water emptying unit to work, the water enriched with the sample is emptied, and after the water is emptied, the water biological sample collection is completed;
D. and (3) water emptying: after the work of the collection unit is finished, the main control board (10) controls the water emptying unit to work, the vacuum pump (6) is started to empty the residual liquid in the enrichment collector, after the water is emptied, one working process is finished, and the next sampling can be performed by replacing the collector (4);
E. decompression: if the blockage phenomenon occurs in the sample collection process, the pipeline pressure can be excessively large, and at the moment, if the pipeline pressure is larger than the critical pressure value of the pressure relief valve (19), the pressure relief valve (19) is automatically opened to form a liquid circulation pipeline, and the pipeline pressure is regulated to effectively protect the device.
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