CN115725390A - System for full-automatic sectional type of normal position is enriched water environment DNA - Google Patents
System for full-automatic sectional type of normal position is enriched water environment DNA Download PDFInfo
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Abstract
The invention relates to an in-situ full-automatic sectional type water environment DNA enrichment system which comprises a water sample suction head, a primary separation device, a secondary separation device, a tertiary separation device, a DNA enrichment device, a circulating water container, a plankton sample collection device and a central controller. The system can obtain four biological indexes which are crucial to water environment evaluation in one water sample treatment: including water-sensitive zooplankton, phytoplankton, planktonic bacteria, and free DNA (the major component is macrophyte DNA). The system can be arranged at a preselected water area monitoring point, so that a water sample is taken regularly for enriching the environmental DNA within one year or even longer, the volume of the environmental DNA sample obtained by enrichment is small, the environmental DNA sample is convenient to carry, and the environmental DNA sample can be easily transferred to a laboratory for relevant detection and analysis. Therefore, the biodiversity condition in the target water area can be monitored for a long time, and the biodiversity condition can be taken as one of the water environment monitoring factors to be brought into the water ecology evaluation system.
Description
Technical Field
The present invention relates to the field of water ecology research and monitoring, and more particularly, to a system that can be used to enrich the DNA of a water environment.
Background
The water ecological health diagnosis is an important ring for environmental protection, treatment and recovery. Aquatic organisms are important indicators for evaluating water environments. Because the plankton in the water, especially the larger plankton, is not uniformly distributed, even the collection of a water sample with a larger volume for plankton analysis is still insufficient for complete statistics of the plankton in the water body. To compensate for this problem, environmental DNA (eDNA) in water samples is often also analyzed in water environmental monitoring. Environmental DNA in water bodies includes free DNA and in vivo DNA in plankton. The detection and analysis of environmental DNA in a body of water can be used to indicate the composition of species in the body of water, characterizing the health of the water ecosystem.
At present, environmental DNA is obtained by collecting plankton in a water body and then cracking and extracting DNA, but the mode ignores free DNA composition in the environment, and the part is mainly used for diversity monitoring and evaluation of fishes in the water body and the like. DNA enrichment materials can be used to enrich for free DNA in water, but because the concentration of free DNA in water is inherently very low. Similarly, the existing water environment DNA enrichment device can not collect plankton with low abundance well. To obtain the DNA and plankton, a large amount of water samples need to be enriched, but the large-volume water samples are transported back to a laboratory for enrichment, which is inconvenient and causes pollution in the transportation process. Further, if the water for enriching free DNA contains too many particles (for example, plankton), the enriching effect and the service life of the DNA enriching apparatus are affected.
Therefore, it is necessary to construct an in-situ full-automatic sectional type system for enriching the DNA of the water body environment.
Disclosure of Invention
In order to solve the problems, the invention provides an in-situ full-automatic sectional type system for enriching the DNA of a water environment, which comprises a water sample suction head, a primary separation device, a secondary separation device, a tertiary separation device, a DNA enrichment device, a circulating water container, a plankton sample collection device and a central controller, wherein the water sample suction head is connected with the primary separation device;
the water outlet of the water sample suction head is connected with a water inlet of the first-stage separation device, the filtrate outlet of the first-stage separation device is connected with the water inlet of the second-stage separation device, the filtrate outlet of the second-stage separation device is connected with the water inlet of the third-stage separation device, the filtrate outlet of the third-stage separation device is connected with the water inlet of the DNA enrichment device, one water outlet of the DNA enrichment device is connected with the water inlet of the circulating water container, and one water outlet of the circulating water container is connected with one water inlet of the first-stage separation device;
the central controller is in communication connection with the water sample suction head, the first-stage separation device, the second-stage separation device, the third-stage separation device, the DNA enrichment device, the circulating water container and the plankton sample collection device.
By the system, a large amount of water samples can be concentrated and enriched into plankton concentrated solution and free DNA enriched solution with extremely small volumes in situ, and the plankton concentrated solution and the free DNA enriched solution can be conveniently conveyed to a conditional laboratory for further analysis, so that complete environmental DNA information and plankton classification information are obtained, and the system is an excellent water environment monitoring tool. Free DNA is extracted through large-volume enrichment, potential influences on an ecosystem caused by fish capture and the like are avoided, and a non-damage monitoring method is provided for diversity diagnosis of large aquatic organisms.
The system can be arranged at a preselected water area monitoring point, so that a water sample is periodically taken for enriching the environmental DNA within one year or even longer, the volume of the environmental DNA sample obtained by enrichment is small, the environmental DNA sample is convenient to carry, and the environmental DNA sample can be easily transferred to a laboratory for relevant detection and analysis after each or a plurality of times of enrichment is finished. Therefore, the biodiversity condition in the target water area can be monitored for a long time, and the biodiversity condition can be taken as one of the factors of the water environment and incorporated into a water ecology evaluation system.
In a particular embodiment, the primary separation device comprises a housing and a filter assembly;
the shell is made of a water-tight material and wraps the filter assembly in the inner space;
the filter assembly comprises a cylindrical filter area and a motor, the motor is arranged on the inner wall of the top of the shell and is connected with the outer wall of the top of the filter part, the motor can drive the filter area to rotate by rotating, and the side wall of the filter area is of a screen mesh structure;
a water inlet is formed in the center of the top of the filtering area, penetrates through the shell and is respectively connected with the water sample suction head and the circulating water container through pipelines;
a concave enrichment area is arranged at the bottom of the filtering area, a plankton concentrated solution outlet is arranged at the bottom of the enrichment area, and the plankton concentrated solution outlet penetrates through the shell and is connected with the plankton sample collecting device through a pipeline;
and a filtrate outlet is also formed in the bottom of the shell and is connected with a water inlet of the secondary separation device through a pipeline. The aperture of the side wall screen structure of the filtration zone is set to 20-64 μm, where 64 μm is the size of the plankton mesh commonly used for water quality aquatic organism monitoring (mesh No. 25), and the majority of plankton in normal water is contained in this fraction. For water with low content of particulate matter, the aperture can be increased to 20 μm, so that the high separation performance of the transverse flow field is fully exerted, and the secondary separation pressure is reduced. Wherein 64 mu m is the size of a plankton net (No. 25 net) commonly used for monitoring water quality aquatic organisms. The separation principle of the stage adopts cross-flow field separation, and the performance is high. For water with low content of particulate matter, the pore diameter can be improved to 20 μm, so that the cross flow field separation performance is fully exerted, and the second-stage separation pressure is reduced.
In a specific embodiment, the secondary separation device and the tertiary separation device are provided with an ultrafiltration device of a back washing part, and the filtration pore size of the secondary separation device is larger than that of the tertiary separation device. Preferably, the filter pore size of the second-stage separation device is 0.7-1.2 μm, and the filter pore size of the third-stage separation device is 0.2 μm. Wherein, 0.7 μm and 1.2 μm respectively correspond to the most common filter membranes for water quality test: pore size of GF/F and GF/C glass fiber membranes.
Because the filtration aperture of tertiary separator is 0.2 mu m, and this aperture is the same with the filter membrane aperture of present filtration enrichment bacterium, so, synthesize first order, second grade enrichment sample, zooplankton, phytoplankton and planktonic bacteria in this system can hold back the water sample, do not omit genetic information. In order to ensure the operation efficiency of the system, the system is provided with a primary separation device at the upstream of a secondary separation device and a tertiary separation device, the filter pore diameter of the separation device is 20-64 mu m, and zooplankton and phytoplankton larger than the size are intercepted and collected, thereby ensuring the efficient operation of the downstream secondary separation device and the downstream tertiary separation device.
In a particular embodiment, the plankton sample collection device comprises three plankton sample collection bottles, each connected to a primary separation device, a secondary separation device, and a tertiary separation device, respectively.
We collected the plankton retained by the three separation devices separately, and the first separation device retained all plankton larger than 20-64 μm, where 64 μm is the size of the plankton net (net No. 25) commonly used for water quality aquatic organism monitoring, and most plankton in normal water body is contained in the fraction. Some zooplankton that first order separator intercepted can be as the sensitive index of water quality evaluation. For example, daphnia magna and the like are sensitive to water quality, so plankton separated by the primary separation device can be used as an early warning index of water quality. In addition, the arrangement of the first separation device not only provides a grading effect for plankton in water, but also intercepts most plankton for the following two-stage ultrafiltration grading, prevents the ultrafiltration membrane of the last two-stage separation device from being blocked, and ensures the efficient operation of the last two-stage separation device and the whole system.
The size of plankton intercepted by the secondary separation device is more than 0.7-1.2 μm and less than the first stage interception aperture (20-64 μm). Wherein, 0.7 μm and 1.2 μm respectively correspond to the filter membranes commonly used in water quality test: pore size of GF/F and GF/C glass fiber membranes. The part of the plankton is mainly (ultra) micro-phytoplankton, which comprises a plurality of small, fast-breeding and easily-dispersing phytoplankton, and a plurality of such phytoplankton can cause water bloom outbreak, so that the plankton trapped by the secondary separation device provides important reference for the possibility of water bloom in the future of the water body and the type of the water bloom.
The phytoplankton retained by the tertiary separation device is primarily a few bacteria and may include a proportion of ultramicro-vegetation. The bacteria are sensitive to the organic matter content in water, and if the organic matter content in the water is too high, the bacteria breed in large quantities, so that the water quality is blackened and smelled to form black smelly water. Therefore, plankton intercepted by the three-stage separation device can be used for auxiliary evaluation of the organic matter content in water and can be used as an early warning index of black smelly water.
Free DNA is extracted through large-volume enrichment, potential influences on an ecosystem caused by fish capture and the like are avoided, and a non-damage monitoring method is provided for diversity diagnosis of large aquatic organisms.
Through the arrangement, four biological indexes which are crucial to water environment evaluation can be obtained in one water sample treatment: including water-sensitive zooplankton, phytoplankton, organic-sensitive bacteria, and free DNA (major component is macrophyte DNA). Because the four biological indexes are collected separately, other items cannot be diluted due to overhigh abundance of one of the four biological indexes, the ecological condition of the water environment is better reflected, and important reference is provided for monitoring and early warning of the water environment.
In a specific embodiment, the DNA enrichment device comprises a DNA enrichment tube, an eluent storage container, a DNA sample collection bottle and a balance solution storage container, and the filtrate outlet of the tertiary separation device, the eluent storage container and the balance solution storage container are all connected with the water inlet of the DNA enrichment tube, and the water outlet of the DNA enrichment tube is connected with the DNA sample collection bottle and the circulating water container.
In a specific embodiment, the DNA enrichment device comprises a plurality of DNA enrichment tubes.
In a specific embodiment, the water inlet of the water sampling suction head is provided with a screen with the aperture of 3 mm. The size of the copepods is 3mm, and the 3mm aperture does not influence the enrichment of larger plankton on the one hand, and on the other hand, can prevent large objects from entering the system and influencing the operation of the system.
In a specific embodiment, a detector for detecting the cell density is arranged on the water sample suction head, and the detector is in communication connection with the central controller. When detecting that the cell density in the water is too high, stopping water sample absorption and treatment and protecting the system from being damaged.
Drawings
Fig. 1 is a schematic structural diagram of a system of embodiment 1.
Fig. 2 is a schematic configuration diagram of a primary separation device of the system of embodiment 1.
FIG. 3 is a schematic diagram of the structure of a DNA enrichment device in one embodiment of the system of the present invention.
Fig. 4 is a schematic structural diagram of the system of embodiment 2.
FIG. 5 is a schematic view of the structure of the DNA enrichment apparatus in example 2.
FIG. 6 is a schematic diagram of a water sampling head with an anti-blocking structure, wherein the water sampling head is in a normal water sampling working state.
Fig. 7 is a schematic diagram of the structure of the water sample suction head with an anti-blocking structure, and the water sample suction head is in a state that the filter screen is blocked, and the ultrasonic generator is in a working state.
The part names corresponding to the number numbers in the figures are as follows: 1. a primary separation device, 11, a housing, 11, a filtrate outlet, 12, a filter assembly, 121, a filter area, 122, a motor, 1211, a water inlet, 1212, an enrichment area, 1213, a plankton concentrate outlet, 2, a secondary separation device, 21, a secondary ultrafiltrate outlet, 22, a secondary backwash liquid outlet, 3, a tertiary separation device, 31, a tertiary ultrafiltrate outlet, 32, a tertiary backwash liquid outlet, 4, a DNA enrichment device, 41, a DNA enrichment tube, 411, a DNA enrichment packing, 42, an eluate storage container, 43, a DNA sample collection bottle, 44, a balance liquid storage container, 5, a circulating water container, 6, a plankton sample collection device, 61, a first plankton collection bottle, 62, a second plankton collection bottle, 63, a third plankton collection bottle, 7, a water sample suction head, 70, a suction head main body, 71, a cell density monitoring device, 72, a 10mm aperture filter screen, 73, a 3mm aperture filter screen, 74, a spring telescopic column, 75, a baffle, 76, an ultrasonic generator, and an ultrasonic control switch.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.
Example 1
As shown in FIG. 1, the system of the present embodiment comprises a water sampling tip 7, a primary separating device 1, a secondary separating device 2, a tertiary separating device 3, a DNA enriching device 4, a circulating water container 5, a plankton sample collecting device 6, and a central controller.
The water outlet of the water sample suction head 7 is connected with a water inlet of the first-stage separation device 1, the filtrate outlet of the first-stage separation device 1 is connected with the water inlet of the second-stage separation device 2, the filtrate outlet of the second-stage separation device 3 is connected with the water inlet of the third-stage separation device 3, the filtrate outlet of the third-stage separation device 3 is connected with the water inlet of the DNA enrichment device 4, a water outlet of the DNA enrichment device 4 is connected with the water inlet of the circulating water container 5, and a water outlet of the circulating water container 5 is connected with a water inlet of the first-stage separation device 1.
The stainless steel mesh screen with the aperture being more than 3mm is arranged at the water inlet of the water sample suction head 7, the copepods are the largest in four major zooplankton classes, generally reaching 3mm, the 3mm aperture is used, on one hand, enrichment of larger planktons is not influenced, and on the other hand, large objects can be prevented from entering the system and influencing the operation of the system.
As shown in fig. 2, the primary separating apparatus 1 includes a housing 11 and a filter assembly 12. The housing 11 is made of a watertight material, and includes the filter assembly 12 in an inner space. The filtering assembly 12 includes a cylindrical filtering area 121 and a motor 122, the motor 122 is disposed on the inner wall of the top of the housing 11 and connected to the outer wall of the top of the filtering area 121, and the motor 122 rotates to drive the filtering area 121 to rotate. The side wall of the filtering area 121 is provided with a screen mesh structure with the aperture of 20-64 mu m, the bottom of the filtering area 121 is provided with a concave enrichment area 1212, and the enrichment area 1212 has the volume of 5-50 mL. The central top of the filtering area 121 is provided with a water inlet 1211 which passes through the shell 11 and is respectively connected with the water sample suction head 7 and the circulating water container 5 through pipelines. The bottom of the enrichment region 1212 is provided with a plankton concentrate outlet 1213, and the plankton concentrate outlet 1213 passes through the housing 11 and is connected with the plankton sample collection device 6 through a pipeline. The bottom of the shell 11 is also provided with a filtrate outlet 111, and the filtrate outlet 111 is connected with the water inlet of the second-stage separation device 2 through a pipeline.
The two-stage separation device 2 and the three-stage separation device 3 are ultrafiltration devices with a back washing part, and are different in that the aperture of a filter membrane of the two-stage separation device 2 is set to be 0.7-1.2 mu m, and the aperture of a filter membrane of the three-stage separation device 3 is set to be 0.2 mu m.
The second grade separator 2 is provided with a second grade ultrafiltrate outlet 21 and a second grade backwash liquid outlet 22, the second grade ultrafiltrate outlet 21 is connected with the water inlet of the third grade separator through a pipeline, and the second grade backwash liquid outlet 22 is connected with the plankton sample collection device 6 through a pipeline.
The third-stage separation device 3 is provided with a third-stage ultrafiltrate outlet 31 and a third-stage backwash liquid outlet 32, the third-stage ultrafiltrate outlet 31 is connected with the water inlet of the DNA enrichment device 4 through a pipeline, and the third-stage backwash liquid outlet 32 is connected with the plankton sample collection device 6 through a pipeline.
As shown in FIG. 3, the DNA enrichment apparatus 4 includes a DNA enrichment tube 41, an eluent storage container 42, a DNA sample collection bottle 43, and a equilibration liquid storage container 44. The DNA enrichment tube 41 is filled with a DNA enrichment packing 411. The filtrate outlet 111 is connected to an inlet of the DNA enrichment tube 41. The eluent reservoir 42 and the equilibrium solution reservoir 44 are connected to an inlet of the DNA enrichment tube 41, and the DNA sample collection bottle 43 is connected to an outlet of the DNA enrichment tube 41.
One water outlet of the DNA enrichment pipe 41 is connected with the circulating water container 5, and the treated water is pumped into the circulating water container 5 by a pump for temporary storage. A water outlet of the circulating water container 5 is connected with a water inlet 1211 of the primary separating device 1, and water can be supplied to the primary separating device 1.
All the water inlets, the water outlets, the waste liquid outlets and the like are provided with valves at necessary positions to control opening and closing, all the valves and the pumps are electrically connected with a central controller, and the working states of the valves and the pumps are controlled by the central controller.
When the device is used, the water sampling suction head 7 sucks a water sample and feeds the water sample into the filtering part 121 of the primary separation device 1, the motor 122 drives the filtering area 121 to rotate to form a transverse flow field in the feeding process, centrifugal force is provided to enable the water sample in the filtering area 121 to pass through micropores in the side wall of the filtering area 121, separated plankton flows out from the plankton concentrated solution outlet 1213 and enters the plankton collection device 6, and obtained primary filtrate flows out from the filtrate outlet 111 of the shell and enters the secondary separation device 2. The primary filtrate is further filtered in the secondary separation device 2, the separated plankton flows out from the secondary backwash liquid outlet 22 into the plankton collection device 6, and the obtained secondary filtrate flows out from the secondary filtrate outlet 21 into the tertiary separation device 3. The secondary filtrate is further filtered in the tertiary separation device 3, the separated plankton flows out from the tertiary backwash liquid outlet 32 and enters the plankton collection device 6, and the obtained tertiary filtrate flows out from the tertiary filtrate outlet 31 and enters the DNA enrichment device 4. The third filtrate adsorbs the DNA in the water to the DNA-rich packing 411 after passing through the DNA-rich apparatus 4. The treated water flows out through a water outlet of the DNA enrichment pipe 41 and is temporarily stored in the circulating water container 5. After the water sample treatment is completed, part of the water stored in the circulating water container 5 can be supplied to the primary separation device 1 to wash the primary separation device 1, so that the primary separation device 1 can be recycled. The equilibrium liquid storage container 44 in the DNA enrichment device 4 supplies the equilibrium liquid to the DNA enrichment tube, and recovers the DNA adsorption function of the DNA enrichment material 411, so that the DNA enrichment device 4 can be recycled. The second-stage separation device 2 and the third-stage separation device 3 are ultrafiltration devices with a back washing part, have washing functions and can be recycled. Therefore, the system of the present embodiment is a recyclable system, and can automatically operate under the control of the central controller.
Through the system of this embodiment, can concentrate the enrichment with a large amount of water samples into plankton concentrate and the free DNA enrichment solution of minimum volume on line, both can conveniently be transported to the conditional laboratory and carry out further analysis to obtain complete environment DNA information, and plankton classification information, be the fine water environment monitoring tool.
Because tertiary separator's filtration aperture is 0.2 mu m, so, synthesize first order, second grade and third grade, this system can hold back zooplankton, phytoplankton and planktonic bacteria in the water sample to collect, with the assurance obtain more comprehensive plankton, do not omit genetic information. In order to ensure the operation efficiency of the system, the system is provided with a primary separation device at the upstream of a secondary separation device and a tertiary separation device, the filtering pore diameter of the separation device is 20-64 mu m, zooplankton and phytoplankton larger than the size are intercepted and collected, and the high-efficiency operation of the downstream secondary separation device is ensured. Therefore, the system of the embodiment can be arranged at a preselected water area monitoring point, so that a water sample is taken regularly for enriching the environmental DNA in one year or even longer, the volume of the environmental DNA sample obtained by enrichment is small, the environmental DNA sample is convenient to carry, and the environmental DNA sample can be easily transferred to a laboratory for relevant detection and analysis. Therefore, the biodiversity condition in the target water area can be monitored for a long time, and the biodiversity condition can be taken as one of the factors of the water environment and incorporated into a water ecology evaluation system.
Example 2
In order to make the system of the present invention work more effectively, we improve on the basis of embodiment 1.
As shown in fig. 4, the plankton sample collection apparatus 6 includes a first plankton collection bottle 61, a second plankton collection bottle 62, and a third plankton collection bottle 63. The first plankton collecting bottle 61 is connected with the plankton concentrated liquid outlet 1213 of the first-stage separation device 1, the second plankton collecting bottle 62 is connected with the second-stage backwash liquid outlet 22, and the third plankton collecting bottle 63 is connected with the third-stage backwash liquid outlet 32. And, each plankton collection bottle is pre-filled with a fixative to maintain the form of plankton entering the collection bottle and prevent it from spoiling.
Through the arrangement, plankton samples intercepted by the three separation devices are separately stored, and the three separated samples respectively have index significance due to the arrangement of the filter pore size.
First, the primary separation device 1 retains all plankton larger than 20-64 μm, where 64 μm is the size of the plankton net commonly used for water quality aquatic organism monitoring (net No. 25), and the vast majority of zooplankton and phytoplankton in normal water bodies are contained in this fraction. Moreover, plankton of this size is also relatively easy to observe with an optical microscope. Some zooplankton intercepted by the primary separation device can be used as a sensitive index for water quality evaluation. For example, daphnia magna and the like are sensitive to water quality, so plankton separated by the primary separation device can be used as an early warning index of water quality. In addition, the arrangement of the first separation device not only provides a grading effect for plankton in water, but also intercepts most plankton for the following two-stage ultrafiltration grading, prevents the ultrafiltration membrane of the last two-stage separation device from being blocked, and ensures the efficient operation of the last two-stage separation device and the whole system.
The size of plankton retained by the secondary separation device 2 is larger than 0.7-1.2 μm and smaller than the first-stage retention pore size (20-64 μm). Wherein, 0.7 μm and 1.2 μm respectively correspond to the most common filter membranes for water quality test: pore size of GF/F and GF/C glass fiber membranes. The part of the plankton is mainly (ultra) micro-phytoplankton which comprises a plurality of small, fast-breeding and easily-spreading phytoplankton, and many of the phytoplankton can cause the outbreak of the water bloom, so that the plankton trapped by the secondary separation device 2 provides important references for the possibility of the future occurrence of the water bloom and the type of the water bloom.
The plankton retained by the tertiary separation device 3 is mainly some bacteria and part of the ultra-micro phytoplankton. The bacteria are sensitive to the organic matter content in water, and if the organic matter content in the water is too high, the bacteria breed in large quantities, so that the water quality is blackened and smelled to form black smelly water. Therefore, the plankton fraction trapped by the third-stage separation device 3 can be used for auxiliary evaluation of the organic matter content in water and as an early warning index of black smelly water.
In one embodiment, a phytoplankton concentration monitoring device 61 is provided on the watersampling tip 6 to monitor the phytoplankton concentration at the site where the watersampling tip 6 is located. The phytoplankton concentration monitoring device 61 is in communication connection with the central controller and sends phytoplankton concentration data to the central controller.
In the practical process, the concentration of the plankton in some water bodies is greatly changed, and when water bloom occurs, a plurality of dominant phytoplankton breed greatly, so that a large amount of plankton is contained in the water. If the device of example 1 is used, a large amount of dominant phytoplankton DNA enters the total DNA collected, resulting in a large dilution of other environmental DNA in the water, which is not favorable for the statistics of the overall biodiversity in the water body.
When the cell density monitoring device is used, before the water sample sucking head 7 starts sucking a water sample, the cell density monitoring device 71 detects the cell density at the water sample sucking head 7 and sends cell density data to the central controller, and the central controller controls the operation mode of the system according to the cell density. When the cell density is low, the system periodically sucks a water sample from water to enrich plankton and free DNA.
When water bloom or black and odorous water is burst in a water area, the concentration of phytoplankton cells or bacteria cells breaks through a threshold value, and the central controller stops water sample absorption in time to prevent the system from being damaged.
In this embodiment, the DNA enrichment apparatus is further optimized, as shown in FIG. 5, a plurality of DNA enrichment tubes 41 are provided to improve the DNA enrichment efficiency.
Example 3
The water sample suction head 7 filters large objects by adopting a 3mm filter screen, but the aperture of 3mm is small, so that blockage is easy to occur. Therefore, in order to solve the problem, the water sampling suction head 7 is improved and designed. As shown in FIGS. 6 and 7, the water sampling tip 7 of the present embodiment comprises a tip body 70, which is a container with an open bottom, which is closed by a filter 72 with a 10mm pore size, and the top of the tip body 70 is connected to a water pipe so that the inside of the tip body 70 can communicate with a pipe pump system of the system. The suction head main body 70 is fixed with a 3mm filter screen 73 at the position close to the top, a spring telescopic rod 74 is fixed at the bottom of the 3mm filter screen 73, the other end of the spring telescopic rod 74 is fixedly connected with a baffle plate 75, and the shape of the baffle plate 75 is matched with the bottom opening of the suction head main body 70, so that the bottom opening of the suction head main body 70 can be just blocked. An ultrasonic generator 76 is provided on the inner wall of the tip body 70, and the ultrasonic generator 76 is provided below the 3mm aperture filter. An ultrasonic generator switch 761 is provided at the bottom opening of the suction head body 70 above the 10mm aperture filter 72.
When the spring telescopic rod 74 is set to be in a natural state, the baffle 75 is blocked, and when the water sample suction head 7 starts to suck water samples by the pump, the spring telescopic rod 74 is contracted by the negative pressure formed by the suction head main body 70, the baffle 75 is lifted, the water samples enter from the bottom opening of the suction head main body 70, and larger floating objects are blocked by the 10mm aperture filter screen 72 and are filtered by the 3mm aperture filter screen 73 in the suction head main body 70 to enter the system. When the filter net 73 with the aperture of 3mm is blocked, the negative pressure in the suction head main body 70 gradually decreases to cause the telescopic spring rod 74 to extend under the thrust of the spring, the baffle plate 75 decreases to seal the bottom opening of the suction head main body 70, and the ultrasonic generator switch 761 is touched to enable the ultrasonic generator 76 to be in a working state, the ultrasonic generator generates ultrasonic waves in the water in the suction head main body 70, so that foreign matters blocked on the filter net 73 with the aperture of 3mm are separated, the filter net 73 with the aperture of 3mm is dredged, and water sample absorption is continued.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and should not be taken as limiting the scope of the present invention, which is intended to cover any modifications, equivalents, improvements, etc. within the spirit and scope of the present invention.
Claims (10)
1. An in-situ full-automatic sectional type water environment DNA enrichment system is characterized by comprising a water sample suction head, a primary separation device, a secondary separation device, a tertiary separation device, a DNA enrichment device, a circulating water container, a plankton sample collection device and a central controller;
the delivery port of water sample suction head with a water inlet of one-level separator is connected, the filtrating export of one-level separator with the water inlet of second grade separator is connected, second grade separator's filtrating export with third grade separator's water inlet is connected, third grade separator's filtrating export with DNA enrichment facility water inlet is connected, a delivery port of DNA enrichment facility with circulating water container's water inlet is connected, circulating water container's a delivery port with a water inlet of one-level separator is connected, central controller with water sample suction head, one-level separator, second grade separator, third grade separator, DNA enrichment facility and circulating water container and plankton sample collection device communication connection.
2. The system of claim 1, wherein the primary separation device comprises a housing and a filter assembly;
the shell is made of a water-tight material and wraps the filter assembly in the inner space;
the filter assembly comprises a cylindrical filter area and a motor, the motor is arranged on the inner wall of the top of the shell and is connected with the outer wall of the top of the filter area, the motor can drive the filter area to rotate by rotating, and the side wall of the filter area is of a screen mesh structure;
a water inlet is formed in the center of the top of the filtering area, penetrates through the shell and is respectively connected with the water sample suction head and the circulating water container through pipelines;
a concave enrichment area is arranged at the bottom of the filtering area, a plankton concentrated solution outlet is arranged at the bottom of the enrichment area, and the plankton concentrated solution outlet penetrates through the shell and is connected with the plankton sample collecting device through a pipeline;
and a filtrate outlet is also formed in the bottom of the shell and is connected with a water inlet of the secondary separation device through a pipeline.
3. The system of claim 2, wherein the apertures of the sidewall screen structure of the filter section are set at 20-64 μm.
4. The system of claim 1, wherein the secondary separation device and the tertiary separation device are provided with an ultrafiltration device having a backwash section, and the secondary separation device has a larger filtration pore size than the tertiary separation device.
5. The system of claim 4, wherein the secondary separation device has a filter pore size of 0.7-1.2 μm.
6. The system of claim 1, wherein the planktonic sample collection device comprises three planktonic sample collection bottles, each connected to a primary separation device, a secondary separation device, and a tertiary separation device, respectively.
7. The system of claim 1, wherein the DNA enrichment device comprises a DNA enrichment tube, an eluent storage container, a DNA sample collection bottle, and a balance solution storage container, and wherein the filtrate outlet of the tertiary separation device, the eluent storage container, and the balance solution storage container are all connected to the water inlet of the DNA enrichment tube, and the water outlet of the DNA enrichment tube is connected to the DNA sample collection bottle and the circulating water container.
8. The system of claim 7, wherein the DNA enrichment device comprises a plurality of DNA enrichment tubes.
9. The system according to claim 1, wherein the water inlet of the water sampling suction head is provided with a screen with an aperture of 3mm, the water sampling suction head is provided with a detector for detecting the cell concentration, and the detector is in communication connection with the central controller.
10. The system of claim 9, wherein the water sampling tip comprises a tip body, which is an open-bottomed container, the open bottom of the water sampling tip is blocked by a 10mm pore size filter screen, and the top of the tip body is connected with a water pipe, so that the inside of the tip body can communicate with a pipe pump system of the system;
the utility model discloses a suction head, including suction head main part, 3mm filter screen, the fixed spring telescopic link in 3mm filter screen bottom, spring telescopic link's the other end and baffle fixed connection, the shape of baffle with the bottom opening cooperation of suction head main part, be provided with supersonic generator on the inner wall of suction head main part, and supersonic generator sets up the below of 3mm aperture filter screen, the supersonic generator switch sets up suction head main part bottom opening part is located 10mm aperture filter screen top.
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| CN202211363024.4A CN115725390A (en) | 2022-11-02 | 2022-11-02 | System for full-automatic sectional type of normal position is enriched water environment DNA |
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| CN202211363024.4A CN115725390A (en) | 2022-11-02 | 2022-11-02 | System for full-automatic sectional type of normal position is enriched water environment DNA |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116296526A (en) * | 2023-05-18 | 2023-06-23 | 交通运输部天津水运工程科学研究所 | Plankton stable isotope sample collection device |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116296526A (en) * | 2023-05-18 | 2023-06-23 | 交通运输部天津水运工程科学研究所 | Plankton stable isotope sample collection device |
| CN116296526B (en) * | 2023-05-18 | 2023-08-22 | 交通运输部天津水运工程科学研究所 | Plankton stable isotope sample collection device |
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