CN115281140A - Aquatic product biological marker culture method - Google Patents
Aquatic product biological marker culture method Download PDFInfo
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- CN115281140A CN115281140A CN202210966263.2A CN202210966263A CN115281140A CN 115281140 A CN115281140 A CN 115281140A CN 202210966263 A CN202210966263 A CN 202210966263A CN 115281140 A CN115281140 A CN 115281140A
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- 239000000090 biomarker Substances 0.000 title claims abstract description 24
- 238000012136 culture method Methods 0.000 title claims abstract description 14
- 241000196324 Embryophyta Species 0.000 claims abstract description 46
- 239000000463 material Substances 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- PWKNBLFSJAVFAB-UHFFFAOYSA-N 1-fluoro-2-nitrobenzene Chemical compound [O-][N+](=O)C1=CC=CC=C1F PWKNBLFSJAVFAB-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910052747 lanthanoid Inorganic materials 0.000 claims abstract description 12
- 150000002602 lanthanoids Chemical class 0.000 claims abstract description 12
- 229910052751 metal Inorganic materials 0.000 claims abstract description 12
- 239000002184 metal Substances 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 235000002732 Allium cepa var. cepa Nutrition 0.000 claims abstract description 10
- 238000009360 aquaculture Methods 0.000 claims abstract description 10
- 244000144974 aquaculture Species 0.000 claims abstract description 10
- 238000009395 breeding Methods 0.000 claims abstract description 9
- 238000011534 incubation Methods 0.000 claims abstract description 8
- AWDWVTKHJOZOBQ-UHFFFAOYSA-K europium(3+);trichloride;hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Cl-].[Eu+3] AWDWVTKHJOZOBQ-UHFFFAOYSA-K 0.000 claims abstract description 7
- ULJUVCOAZNLCJZ-UHFFFAOYSA-K trichloroterbium;hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Cl-].[Tb+3] ULJUVCOAZNLCJZ-UHFFFAOYSA-K 0.000 claims abstract description 7
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 241000234282 Allium Species 0.000 claims abstract 2
- 241000252212 Danio rerio Species 0.000 claims description 15
- 241000251468 Actinopterygii Species 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 2
- 231100000956 nontoxicity Toxicity 0.000 abstract description 4
- 239000003550 marker Substances 0.000 abstract description 3
- 230000004071 biological effect Effects 0.000 abstract description 2
- 238000000227 grinding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract 1
- 244000291564 Allium cepa Species 0.000 description 13
- 239000000126 substance Substances 0.000 description 11
- 239000008367 deionised water Substances 0.000 description 8
- 229910021641 deionized water Inorganic materials 0.000 description 8
- 238000002372 labelling Methods 0.000 description 6
- 238000002073 fluorescence micrograph Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241001092371 Bergenia Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- VIQSRHWJEKERKR-UHFFFAOYSA-L disodium;terephthalate Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=C(C([O-])=O)C=C1 VIQSRHWJEKERKR-UHFFFAOYSA-L 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000003777 experimental drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001215 fluorescent labelling Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000003578 releasing effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/90—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination
- A01K61/95—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination specially adapted for fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
- C08G83/008—Supramolecular polymers
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/182—Metal complexes of the rare earth metals, i.e. Sc, Y or lanthanide
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Marine Sciences & Fisheries (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Health & Medical Sciences (AREA)
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Abstract
The invention provides an aquatic product biomarker culture method. The method comprises the steps of firstly placing living plants in simulated sunlight, incubating the living plants in a disodium terephthalate solution, transferring the living plants to a lanthanide metal solution for continuous incubation, freeze-drying the living plants to obtain the plant interface super-assembled SAFs fluorescent material, grinding the plant interface super-assembled SAFs fluorescent material, and feeding the ground SAFs fluorescent material to a water product for aquaculture biological marker culture. The selected living plant is green onion, and the lanthanide metal is europium chloride hexahydrate or terbium chloride hexahydrate. According to the method for breeding the aquatic organism marker, provided by the invention, as the selected plant has no toxicity and the prepared plant interface super-assembly SAFs fluorescent material also has no toxicity, the plant interface super-assembly SAFs fluorescent material can be completely metabolized within 24 hours without influencing the biological activity of aquatic organisms.
Description
Technical Field
The invention belongs to the technical field of aquatic product marker culture, and particularly relates to an aquatic product biomarker culture method.
Background
With the increasing development of aquaculture and the increasing demand for fishery resource conservation, the technology of fish marking has made great progress, and various marking methods have been produced. However, these methods have many problems and disadvantages in labeling larval fish, such as requiring a long time or high economic cost, and each labeling method has all or all of such disadvantages and disadvantages due to the reason for labeling, the strength of labeling, and the identification of labeling. The stress and fatality rate of the fish body, the marking cost and labor intensity, the detection and recapture effect of the mark and the like are important factors influencing the marking technology, and the fluorescent dye soaked mark can meet the requirements and can realize large-scale mark releasing activity in a short time.
However, at present, in domestic research, when the fluorescent substance is used for soaking and marking aquatic organisms, the concentration of the fluorescent substance is selected more heavily, the soaking time is ignored, and particularly when the fluorescent substance is used for soaking and marking fishes, whether stress, damage and damage degree exist to the organisms or not is not clear.
The fluorescent substance is directly fed into the water product, whether the fluorescent substance is ingested or not is judged by the aquatic organisms through signal identification, the fluorescent substance is not ingested due to the fact that the aquatic organisms do not ingest the water product after the signal identification of the fluorescent substance is different from a food signal, and the fluorescent substance cannot enter the water product and cannot play a role in marking.
Disclosure of Invention
Aquatic organisms generally feed on plants or plankton, according to the characteristics, fluorescent substances are super-assembled on the internal interface of the plants fed by the aquatic organisms, the signals of the aquatic organisms identify the fed plants, and the aquatic organisms and the fluorescent substances are taken into the bodies when the plants are taken.
In order to solve the problems of the prior art, the invention provides an aquatic biomarker culture method.
The specific technical scheme of the invention is as follows:
the invention provides an aquatic product biomarker culture method which is characterized by comprising the following steps: the plant interface super-assembled SAFs fluorescent material is crushed and fed to a water production product, wherein the preparation method of the plant interface super-assembled SAFs fluorescent material comprises the following steps: placing living plants in simulated sunlight, incubating in a disodium terephthalate solution, transferring to a lanthanide metal solution for continuous incubation, and freeze-drying to obtain the plant interface super-assembly SAFs fluorescent material.
The method for culturing the aquatic organism marker provided by the invention is also characterized in that the aquatic organism is fish.
The aquatic product biomarker culture method provided by the invention is also characterized in that the fish is zebra fish.
The aquatic product biomarker culture method provided by the invention is also characterized in that the living plant is green onion, and the lanthanide metal is europium chloride hexahydrate or terbium chloride hexahydrate.
The aquatic product biomarker culture method provided by the invention is also technically characterized in that the concentration of the disodium terephthalate solution is 1-300 mmol/L and the dosage is 0.5-50 ml, and the concentration of the lanthanide metal solution is 1-300 mmol/L and the dosage is 0.5-60 ml.
The aquatic biomarker culture method provided by the invention also has the technical characteristics that the simulated sunlight is generated by a high-power sunlight simulator.
The aquatic product biomarker culture method provided by the invention is also characterized in that the power of the high-power sunlight simulator is 100-3000W.
The aquatic product biomarker breeding method provided by the invention is also technically characterized in that the incubation time in the disodium terephthalate solution is 2-72 h, and the continuous incubation time in the lanthanide metal solution is 2-72 h.
Action and Effect of the invention
The method comprises the steps of firstly placing living plants in simulated sunlight, incubating the living plants in a disodium terephthalate solution, transferring the living plants to a lanthanide metal solution for continuous incubation, freeze-drying the living plants to obtain the plant interface super-assembly SAFs fluorescent material, grinding the plant interface super-assembly SAFs fluorescent material, and feeding the ground plant interface super-assembly SAFs fluorescent material to a water product for aquaculture biological marker culture. The selected living plant is green onion, and the lanthanide metal is europium chloride hexahydrate or terbium chloride hexahydrate.
Therefore, the aquatic biomarker breeding method provided by the invention has the advantages that the selected plants are not toxic, and the prepared plant interface super-assembly SAFs fluorescent material is also not toxic, so that the plant interface super-assembly SAFs fluorescent material can be completely metabolized within 24 hours without influencing the aquatic biological activity.
Drawings
FIG. 1 is a fluorescent image of unlabeled zebrafish under different fluorescent conditions.
Fig. 2 is a fluorescence image of the labeled zebrafish of example 1 under different fluorescence conditions.
FIG. 3 is a 245nm fluorescence image of the labeled zebrafish of example 1 at various times.
FIG. 4 shows Raw246.7 cell viability of the plant interface super-assembled SAFs fluorescent material and the scallion after the Raw246.7 cells are treated for 24 hours.
Detailed Description
Terms used in the present invention generally have meanings commonly understood by those of ordinary skill in the art, unless otherwise specified.
In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.
The reagents used in the following examples are commercially available and the experimental procedures and experimental conditions not specified are those conventional in the art.
The living onions used in the following examples were purchased from the Strong bergenia market in Jinan. Experimental drugs were purchased from aladdin, CAS number disodium terephthalate: 10028-70-3, molecular formula: c 8 H 4 Na 2 O 4 Molecular weight: 210.1; europium chloride hexahydrate CAS number: 13759-92-7, formula: eucl 3 ·6H 2 O, molecular weight: 366.41; terbium chloride hexahydrate CAS number: 13798-24-8, formula: tbCl 3 ·6H 2 O, molecular weight: 373.38.WINSURE brand high power sunlight simulator. PE small animal living body imager. Zebrafish were purchased from Taobao.
The following description of the embodiments of the present invention will be made with reference to the accompanying drawings.
< example 1>
In the embodiment, the plant interface super-assembly SAFs fluorescent material is prepared from terbium chloride hexahydrate and used for zebra fish marking culture, and the specific process is as follows:
adding 840mg of disodium terephthalate into 20mL of deionized water, stirring for 30min, then placing living green onion (SO) into the deionized water, and incubating for 48h under the irradiation of a high-power sunlight simulator (the power is 300W); adding 149mg of terbium chloride hexahydrate into 20mL of deionized water, stirring for 30min, then vertically immersing the living green onion incubated in the disodium terephthalate solution, continuing to incubate for 48h under the irradiation of a high-power sunlight simulator (the power is 300W), finally washing with the deionized water, and freeze-drying in a freeze dryer to obtain the plant interface super-assembly SAFs fluorescent material; the material was crushed and 2g of zebrafish (20 strips) fed on an empty stomach for 2 days were photographed and observed with a PE small animal in vivo imager.
FIG. 1 is a fluorescent image of unlabeled zebrafish under different fluorescent conditions. Fig. 2 is a fluorescence image of the labeled zebrafish of example 1 under different fluorescence conditions.
Comparing fig. 1 and 2, it can be known that the plant interface super-assembly SAFs fluorescent material prepared in this embodiment can indeed perform a fluorescent labeling effect on zebra fish, that is, the aquatic organism labeling culture method provided in this embodiment is indeed feasible.
FIG. 3 is a 245nm fluorescence image of the labeled zebrafish of example 1 at various times. As can be seen from FIG. 3, the fluorescent substance in the body of the labeled zebra fish of the present example was completely metabolized within 24 hours.
< example 2>
In the embodiment, the plant interface super-assembly SAFs fluorescent material is prepared by europium chloride hexahydrate and used for zebra fish marking culture, and the specific process is as follows:
adding 840mg of disodium terephthalate into 20mL of deionized water, stirring for 30min, then placing living green onion (SO) into the deionized water, and incubating for 48h under the irradiation of a high-power sunlight simulator (the power is 300W); adding 1476mg of europium chloride hexahydrate into 20mL of deionized water, stirring for 30min, then vertically immersing the living green onion incubated in the disodium terephthalate solution, continuously incubating for 48h under the irradiation of a high-power sunlight simulator (the power is 300W), finally washing with the deionized water, and freeze-drying in a freeze dryer to obtain the plant interface super-assembled SAFs fluorescent material; the material was crushed and 2g of the material was fed to zebrafish (20) on an empty stomach for 2 days.
FIG. 4 shows Raw246.7 cell viability of the plant interface super-assembled SAFs fluorescent material prepared in the example and after Raw246.7 cells are treated by green onion for 24 hours.
Cell viability was compared with Raw246.7 cells (Blank) without any treatment after treating Raw246.7 cells with the plant interface super assembled SAFs fluorescent material prepared in examples 1 and 2 and Scallion (SO) for 24h, and the results are shown in FIG. 4.
As can be seen from FIG. 4, the plants selected by the aquatic biomarker culture method provided by the invention have no toxicity to Raw246.7 cells, and the prepared plant interface super-assembly SAFs fluorescent material also has no toxicity to Raw246.7 cells.
The foregoing is a detailed description of embodiments that will enable those skilled in the art to make and use the invention. The technical solutions of the present invention, which can be improved or modified only by analysis, analogy or limited enumeration, should be within the scope of protection determined by the claims.
Claims (8)
1. An aquaculture biomarker culture method, comprising the steps of:
the plant interface super-assembled SAFs fluorescent material is crushed and fed to water producing matter,
the preparation method of the plant interface super-assembly SAFs fluorescent material comprises the following steps:
and (2) placing living plants in simulated sunlight, incubating in a disodium terephthalate solution, transferring to a lanthanide metal solution for continuous incubation, and freeze-drying to obtain the plant interface super-assembly SAFs fluorescent material.
2. The aquaculture biomarker breeding method according to claim 1,
wherein the aquatic organism is fish.
3. The method of claim 2 wherein the biomarker is derived from a plant species,
wherein the fish is zebra fish.
4. The aquaculture biomarker breeding method according to claim 1,
wherein the living plant is a green onion,
the lanthanide metal is europium chloride hexahydrate or terbium chloride hexahydrate.
5. The aquaculture biomarker breeding method according to claim 1,
wherein the concentration of the disodium terephthalate solution is 1 to 300mmol/L, the dosage is 0.5 to 50ml,
the concentration of the lanthanide metal solution is 1-300 mmol/L, and the dosage is 0.5-60 ml.
6. The aquaculture biomarker breeding method according to claim 1,
wherein the simulated sunlight is generated by a high-power sunlight simulator.
7. The aquaculture biomarker breeding method according to claim 6,
wherein the power of the high-power solar simulator is 100-3000W.
8. The aquaculture biomarker breeding method according to claim 1,
wherein the incubation time in the disodium terephthalate solution is 2-72 h,
the continuous incubation time in the lanthanide metal solution is 2-72 h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210966263.2A CN115281140A (en) | 2022-08-12 | 2022-08-12 | Aquatic product biological marker culture method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210966263.2A CN115281140A (en) | 2022-08-12 | 2022-08-12 | Aquatic product biological marker culture method |
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| Publication Number | Publication Date |
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| CN115281140A true CN115281140A (en) | 2022-11-04 |
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| CN202210966263.2A Pending CN115281140A (en) | 2022-08-12 | 2022-08-12 | Aquatic product biological marker culture method |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2088903A1 (en) * | 1993-02-04 | 1994-08-05 | Robert Brown | Method for identifying marked fish |
| CN102273418A (en) * | 2011-06-24 | 2011-12-14 | 中国水产科学研究院珠江水产研究所 | Fluorescent labeling method of fish otolith |
| CN107258647A (en) * | 2017-05-24 | 2017-10-20 | 中国水产科学研究院黄海水产研究所 | A kind of lefteye flounder fish otolith rubidium marking method |
| CN107286184A (en) * | 2017-07-10 | 2017-10-24 | 肇庆学院 | A kind of zinc metal-organic framework materials and preparation method thereof |
-
2022
- 2022-08-12 CN CN202210966263.2A patent/CN115281140A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2088903A1 (en) * | 1993-02-04 | 1994-08-05 | Robert Brown | Method for identifying marked fish |
| CN102273418A (en) * | 2011-06-24 | 2011-12-14 | 中国水产科学研究院珠江水产研究所 | Fluorescent labeling method of fish otolith |
| CN107258647A (en) * | 2017-05-24 | 2017-10-20 | 中国水产科学研究院黄海水产研究所 | A kind of lefteye flounder fish otolith rubidium marking method |
| CN107286184A (en) * | 2017-07-10 | 2017-10-24 | 肇庆学院 | A kind of zinc metal-organic framework materials and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| OSEPH J. RICHARDSON等: "Nano-Biohybrids: In Vivo Synthesis of Metal–Organic Frameworks inside Living Plants", 《SMALL》, vol. 14, 23 November 2017 (2017-11-23), pages 1 - 7 * |
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