CN219670495U - Experimental system for ballast water gene sample preparation - Google Patents
Experimental system for ballast water gene sample preparation Download PDFInfo
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- CN219670495U CN219670495U CN202320579562.0U CN202320579562U CN219670495U CN 219670495 U CN219670495 U CN 219670495U CN 202320579562 U CN202320579562 U CN 202320579562U CN 219670495 U CN219670495 U CN 219670495U
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 63
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title claims description 8
- 238000001914 filtration Methods 0.000 claims abstract description 41
- 230000007246 mechanism Effects 0.000 claims abstract description 41
- 239000012528 membrane Substances 0.000 claims abstract description 19
- 239000011521 glass Substances 0.000 claims description 36
- 239000000020 Nitrocellulose Substances 0.000 claims description 11
- 229920001220 nitrocellulos Polymers 0.000 claims description 11
- 238000000967 suction filtration Methods 0.000 claims description 11
- 238000007789 sealing Methods 0.000 claims description 9
- CFOAUMXQOCBWNJ-UHFFFAOYSA-N [B].[Si] Chemical compound [B].[Si] CFOAUMXQOCBWNJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000005388 borosilicate glass Substances 0.000 claims description 2
- 239000012535 impurity Substances 0.000 abstract description 6
- 238000012360 testing method Methods 0.000 abstract description 5
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract description 2
- 230000003115 biocidal effect Effects 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 229940088710 antibiotic agent Drugs 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 241001411320 Eriogonum inflatum Species 0.000 description 2
- 101100223892 Escherichia coli sulI gene Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000000356 contaminant Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000002352 surface water Substances 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000000861 blow drying Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001617 migratory effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Landscapes
- Sampling And Sample Adjustment (AREA)
Abstract
The utility model discloses an experimental system for preparing ballast water gene samples, which comprises a supporting seat, a sample filtering mechanism, a sample enrichment mechanism, a sterile refrigerator and a layered placing rack, wherein the supporting seat is arranged on the supporting seat; the bottom detachable of sample filtering mechanism is provided with sample enrichment mechanism, and sample filtering mechanism includes first order filtration and second grade filtration, and first order filtration gets rid of macroscopic impurity, and second grade filtration gets rid of more than 100 mu m plankton and plant, and sample enrichment mechanism can store the ballast water after the filtration. The experimental system for preparing the ballast water gene sample is easy to detach, sample and clean, convenient to operate, capable of being used in the field or a laboratory, and capable of being sterilized for repeated use, is multistage in filtration arrangement, avoids the blockage of a filter membrane by impurities in the ballast water, can be used for testing different indexes respectively, realizes one-time operation, and reduces resource waste and environmental pollution.
Description
Technical Field
The utility model is suitable for researching the invasion risk of novel pollutants in the process of discharging ballast water of ships, in particular relates to the propagation risk of antibiotic resistance genes in ballast water in marine environment, and is a pretreatment experimental system for extracting antibiotic resistance gene samples in ballast water.
Background
Antibiotics are widely used to protect the health of humans and animals or to increase the growth rate of animals, however 70% of these antibiotics are excreted into the environment in the form of urine and feces, causing the development of drug resistance genes in the environment and further transmitted by organisms. The spread of antibiotic resistance genes (Antibiotic Resistance Genes, ARGs) has become a serious global problem in recent years. In 2006, ARGs were first proposed as an environmentally emerging class of contaminants, unlike other contaminants, ARGs can persist long-lasting in organisms and be transferred to other organisms. Thus, the presence and migratory transformation of ARGs in the environment is more environmentally damaging than the residue of antibiotics themselves.
In recent years, a large number of ARGs have been detected in water environments in estuaries, ports and coastal areas, and the ARGs-containing seawater may be pumped into the ship ballast tanks and transferred to the global ocean. However, the related documents of the existing supervision of the ship ballast water do not have corresponding control requirements on ARGs, and do not have corresponding preparation and detection methods of the ARGs sample of the ship ballast water, so that the ship ballast water becomes an important carrier for the global spread of the ARGs. The system is capable of knowing the ARGs level and composition in the ballast water, is beneficial to grasping and controlling potential adverse effects of the ballast water on marine ecology and human health, and provides basis for ballast water discharge management.
Disclosure of Invention
The utility model is suitable for researching the invasion risk of novel pollutants in the process of discharging ballast water of ships, in particular relates to the propagation risk of antibiotic resistance genes in ballast water in marine environment, and is a pretreatment experimental system for extracting antibiotic resistance gene samples in ballast water.
In order to achieve the above purpose, the present utility model provides the following technical solutions: an experimental system for preparing ballast water gene samples comprises a supporting seat, a sample filtering mechanism, a sample enrichment mechanism, a sterile refrigerator and a layered placing rack;
the bottom end of the sample filtering mechanism is detachably provided with a sample enrichment mechanism and can achieve sealing, the sample filtering mechanism comprises a first-stage filtering and a second-stage filtering, the first-stage filtering removes macroscopic impurities, the second-stage filtering removes zooplankton and plant with the size of more than 100 mu m, and the sample enrichment mechanism can store the ballast water after filtering;
the inner cavity of the sterile refrigerator is provided with a layered placing rack, and the culture dish is placed in the sterile refrigerator and stored by the layered placing rack.
Preferably, a sample filtering mechanism is arranged for realizing double-stage filtering, wherein a second-stage filter cylinder is detachably arranged in the inner cavity of the glass cup main body, and a first-stage filter screen with a conical structure is detachably arranged in the inner cavity of the second-stage filter cylinder.
Preferably, in order to store the filtered ballast water, a sample enrichment mechanism is arranged, the filter head is detachably connected with the glass cup main body, and the water is filtered again by utilizing the sand core filter plate and the 0.22 mu m nitrocellulose filter membrane, so that the water enters an enrichment bottle detachably connected with the bottom end of the filter head.
In the operation process, the two-stage filtering system is built up and put into the glass main body, the glass main body is connected with the enrichment system (through the fixing clamp or the frosted seal), the ballast water samples are filtered through the two stages respectively, then the filtering mechanism can be taken out, the water samples in the glass main body 21 are pumped through the filter membrane of 0.22um, the filter membrane is used for gene extraction, and the water samples can be used for other measurements.
Compared with the prior art, the utility model has the beneficial effects that: the experimental system for preparing the ballast water gene sample is easy to detach, sample and clean, convenient to operate, capable of being used in the field or a laboratory, and capable of being sterilized for repeated use, is multistage in filtration arrangement, avoids the blockage of a filter membrane by impurities in the ballast water, can be used for testing different indexes respectively, realizes one-time operation, and reduces resource waste and environmental pollution.
Drawings
FIG. 1 is a schematic diagram of the structure of the present utility model;
FIG. 2 is a schematic diagram of a combination of a sample filtration mechanism and a sample enrichment mechanism according to the present utility model;
FIG. 3 is a schematic view of a sample filtration mechanism according to the present utility model;
FIG. 4 is a schematic diagram of a sample enrichment mechanism according to the present utility model.
In the figure: 1. the device comprises a supporting seat, 2 parts of sample filtering mechanisms, 21 parts of glass cup bodies, 22 parts of water outlet taps, 23 parts of 100-mu m filter membrane structures, 24 parts of second-stage filter cylinders, 25 parts of clamping rings, 26 parts of first-stage filter screens, 27 parts of clamping rings, 3 parts of sample enrichment mechanisms, 31 parts of enrichment bottles, 32 parts of suction filtration ports, 33 parts of filter heads, 34 parts of sand core filter plates, 35 parts of 0.22-mu m nitrocellulose filter membranes, 4 parts of sterile refrigerators and 5 parts of layered placing racks.
Detailed Description
The following description of the embodiments of the present utility model will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present utility model, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
Referring to fig. 1-4, the present utility model provides a technical solution: the utility model provides an experimental system of ballast water gene sample preparation, includes supporting seat 1, sample filtering mechanism 2, sample enrichment mechanism 3, aseptic fridge 4 and layering rack 5, and the top of supporting seat 1 is provided with sample filtering mechanism 2, sample enrichment mechanism 3 and aseptic fridge 4 from left to right movably in proper order, and wherein, sample filtering mechanism 2 can dismantle the setting on the top of sample enrichment mechanism 3, and the inner chamber of aseptic fridge 4 is provided with layering rack 5.
More specifically, the sample filtration mechanism 2 includes: the glass main body 21, the water outlet tap 22, the 100 mu m filter membrane structure 23, the second-stage filter cylinder 24, the second clamping ring 25, the first-stage filter screen 26 and the first clamping ring 27;
the glass main body 21 is hollow up and down and is made of high boron silicon, the volume is 500ml, the diameter of the bottom edge of the glass main body 21 is larger than the hollow diameter, the edge is made of frosted material, the second-stage filter cylinder 24 is detachably arranged in the inner cavity of the glass main body 21, the diameter of the second-stage filter cylinder 24 is smaller than that of the glass main body 21, the top end of the second-stage filter cylinder 24 is fixedly provided with a clamping ring II 25, the clamping ring II 25 can be sleeved on the top end of the glass main body 21 and used for fixing the second-stage filter cylinder 24 in the glass main body 21, the 100 mu m filter membrane structure 23 is in threaded connection with the bottom end of the second-stage filter cylinder 24, floating animals and plants with the diameter larger than 100 mu m are filtered and removed, the space between the second-stage filter cylinder 24 and the glass main body 21 is the ballast water storage space 2, the first-stage filter cylinder 26 is detachably arranged in the inner cavity of the second-stage filter cylinder 24, the top end of the first-stage filter cylinder 26 is fixedly provided with a clamping ring I27, and the clamping ring I27 can be sleeved on the top end of the second-stage filter cylinder 24 and used for fixing the first-stage filter cylinder 26 in the second-stage filter cylinder 24;
in this embodiment, the first stage filter 26 has a conical structure, and is a large-aperture filter, so as to remove macroscopic impurities;
the space between the first-stage filter screen 26 and the second-stage filter cartridge 24 is the ballast water storage space 1, and the water outlet tap 22 is arranged on the outer wall of the glass main body 21;
annular clamping grooves are formed in the lower surfaces of the first clamping ring 27 and the second clamping ring 25, so that the second clamping ring 25 can be clamped on the glass main body 21 by utilizing the annular clamping grooves, and the first clamping ring 27 can be clamped on the second clamping ring 25 by utilizing the annular clamping grooves.
The ballast water sample is firstly subjected to impurity removal and phytoplankton removal through the sample filtering mechanism 2, the ballast water sample passing through the first-stage filter screen 26 enters the ballast water storage space 1, the partial pressure ballast water can be continuously used for compliance test of the standard of the ballast water convention D-2, the ballast water sample subjected to the second stage filtering through the 100 mu m filter membrane structure 23 enters the ballast water storage space 2, in order to control the outer wall of the suction-filtered volume glass cup main body 21, the side wall of the ballast water storage space 2 is provided with a water outlet tap 22, the water outlet tap is positioned at a 200ml volume marking position of the glass cup, the ballast water volume entering the next link is conveniently controlled, and the ballast water flowing out through the water outlet tap 22 can be continuously used for detecting the content of antibiotics in the ballast water through the procedures of sample extraction, elution, blow drying, re-dissolution and the like.
More specifically, the sample enrichment mechanism 3 includes: an enrichment bottle 31, a suction filtration port 32, a filter head 33, a sand core filter plate 34 and a 0.22 mu m nitrocellulose filter membrane 35;
the enriching bottle 31 is made of high borosilicate glass, the volume is 1L, the inner side and the outer side of the mouth are frosted, a frosted bottle stopper is arranged, the frosted bottle stopper is arranged on the outer side of the mouth and is convenient to be jointed and sealed with the filter head 33, a suction filtration port 32 is arranged at the position 5cm outside the upper part of the bottle body and is used for connecting a suction filtration hose and a motor, the filter head 33 is arranged at the top end of the enriching bottle 31 in a threaded manner, the frosted bottle inside the filter head 33 is arranged, the top of the filter head 33 is arranged in a concentric circle manner, the sand core filter plate 34 is arranged in the inner cavity of the filter head 33, and a 0.22 mu m nitrocellulose filter membrane 35 is paved at the top end of the sand core filter plate 34, and the outer circle diameter of the filter head 33 is equal to the outer edge diameter of the glass cup main body 21, so that the enriching bottle is convenient to be assembled and sealed;
in order to enhance the tightness between the filter head 33 and the glass main body 21, the glass main body 21 and the filter head 33 can be connected by adopting screw threads, and the sealing ring is adopted between the glass main body 21 and the filter head 33 to enhance the effect of fixation and sealing after superposition;
the method comprises the steps of combining a sample filtering mechanism 2 and a sample enriching mechanism 3, placing a 0.22 mu m nitrocellulose filter membrane 35 at a sand core filter plate 34 of a filter head 33 of the sample enriching mechanism 3, firstly, enabling a ballast water sample to pass through a filtering device, removing a two-stage filtering structure after the filtering sample amount is larger than 200ml, carrying out suction filtration on the sample through a motor, removing a system after the suction filtration of 200ml of the sample is finished, wherein the 0.22 mu m nitrocellulose filter membrane 35 is used for detecting ARGs in the ballast water, and storing a wide-mouth bottle mouth and a suction filtration port in a plug manner, wherein a water sample can be used for basic water quality detection.
The sterile refrigerator 4 is used for freezing and preserving the 0.22 mu m nitrocellulose filter membrane 35 after suction filtration, the temperature is set to be minus 20 ℃, the layering rack 5,0.22 mu m nitrocellulose filter membrane 35 which is internally provided with layering storage culture dishes is placed in a sterilization culture dish, the culture dish is placed in the sterile refrigerator, and the sterile refrigerator is brought back to a laboratory, and is used for detecting ARGs after DNA is extracted.
The detailed connection means are known in the art, and the following mainly describes the working principle and process, and the specific work is as follows.
The tightness of the equipment is detected by filtering sterile water, the effectiveness of the equipment is detected by filtering surface water sampled in the field, and three samples are arranged.
Placing the first stage filter screen 26 in the second stage filter cartridge 24, and placing the second stage filter cartridge 24 in the glass body 21 of the filter structure;
the 0.22 mu m nitrocellulose filter membrane 35 is arranged on a sand core filter plate 34 of the filter head 33 and can be wetted by water;
the filter head 33 is assembled and connected with the enrichment bottle 31, and is sealed through a frosted structure;
the glass main body 21 is connected with the filter head 33 connected with the enrichment bottle 31, sealing is achieved through a frosted structure, in order to strengthen the sealing performance between the filter head 33 and the glass main body 21, the glass main body 21 and the filter head 33 can be connected through threads, and sealing rings are adopted between the glass main body 21 and the filter head 33 to strengthen the effect of fixation and sealing after superposition;
the suction port 32 of the enrichment bottle 31 is connected with a motor through a hose;
performing a sealing test on the connected devices through sterile water;
after suction filtration through three different surface water samples, 0.22 μm nitrocellulose was extracted using a DNA extraction kitThe DNA sample on the filter 35 was tested for sul1 concentration. The test results showed that the absolute concentration of sul1 in the three samples was 2.71×10, respectively 5 copies/mL、2.18×10 4 Copies/mL and 2.16X10 4 copies/mL。
Although embodiments of the present utility model have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the utility model, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. The utility model provides an experimental system of ballast water gene sample preparation, its characterized in that, including supporting seat (1), sample filtering mechanism (2), sample enrichment mechanism (3), aseptic fridge (4) and layering rack (5), the top of supporting seat (1) is provided with sample filtering mechanism (2), sample enrichment mechanism (3) and aseptic fridge (4) from left to right movable in proper order, the setting that sample filtering mechanism (2) can dismantle is on the top of sample enrichment mechanism (3) to can reach sealedly, the inner chamber of aseptic fridge (4) is provided with layering rack (5).
2. An experimental system for ballast water gene sample preparation according to claim 1, wherein: the sample filtration mechanism (2) comprises:
a glass main body (21) made of high boron silicon and hollow at the upper and lower sides, wherein the diameter of the bottom edge of the glass main body (21) is larger than the hollow diameter;
the second-stage filter cylinder (24) is detachably arranged in the inner cavity of the glass main body (21), a second clamping ring (25) is fixedly arranged at the top end of the second-stage filter cylinder (24), and the second clamping ring (25) can be sleeved at the top end of the glass main body (21) and is used for fixing the second-stage filter cylinder (24) in the glass main body (21);
a 100 mu m filter membrane structure (23) in threaded connection with the bottom end of the second-stage filter cylinder (24);
the detachable first-stage filter screen (26) is arranged in the inner cavity of the second-stage filter cylinder (24), a first clamping ring (27) is fixedly arranged at the top end of the first-stage filter screen (26), and the first clamping ring (27) can be sleeved at the top end of a second clamping ring (25) and is used for fixing the first-stage filter screen (26) in the second-stage filter cylinder (24);
and a water outlet tap (22) arranged on the outer wall of the glass main body (21).
3. An experimental system for ballast water gene sample preparation according to claim 2, wherein: the diameter of the second stage filter cartridge (24) is smaller than the diameter of the glass body (21).
4. An experimental system for ballast water gene sample preparation according to claim 3, wherein: the first stage filter screen (26) is of a conical structure.
5. The experimental system for preparing a ballast water gene sample according to claim 4, wherein: the sample enrichment mechanism (3) comprises:
the wide-mouth glass bottle is an enrichment bottle (31) made of high borosilicate glass;
a suction filtration port (32) arranged on the outer wall of the enrichment bottle (31) and used for connecting a suction filtration hose and a motor;
a filter head (33) which is arranged at the top end of the enrichment bottle (31) in a threaded manner;
the sand core filter plate (34) is arranged in the inner cavity of the filter head (33) in a threaded manner, and the 0.22 mu m nitrocellulose filter membrane (35) is paved at the top end of the sand core filter plate (34).
6. The experimental system for preparing a ballast water gene sample according to claim 5, wherein: the diameter of the outer circle of the filter head (33) is equal to the diameter of the outer edge of the glass main body (21), so that the assembly and the sealing are convenient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320579562.0U CN219670495U (en) | 2023-03-22 | 2023-03-22 | Experimental system for ballast water gene sample preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320579562.0U CN219670495U (en) | 2023-03-22 | 2023-03-22 | Experimental system for ballast water gene sample preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219670495U true CN219670495U (en) | 2023-09-12 |
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ID=87898283
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320579562.0U Active CN219670495U (en) | 2023-03-22 | 2023-03-22 | Experimental system for ballast water gene sample preparation |
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| Country | Link |
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| CN (1) | CN219670495U (en) |
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- 2023-03-22 CN CN202320579562.0U patent/CN219670495U/en active Active
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