CN114868682A - Shell Cal marking method applied to living bivalve shellfish - Google Patents
Shell Cal marking method applied to living bivalve shellfish Download PDFInfo
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- CN114868682A CN114868682A CN202210231473.7A CN202210231473A CN114868682A CN 114868682 A CN114868682 A CN 114868682A CN 202210231473 A CN202210231473 A CN 202210231473A CN 114868682 A CN114868682 A CN 114868682A
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- Prior art keywords
- shellfish
- mark
- cal
- shell
- marking
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- 235000015170 shellfish Nutrition 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000004043 dyeing Methods 0.000 claims abstract description 10
- 230000005284 excitation Effects 0.000 claims abstract description 9
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229960002378 oftasceine Drugs 0.000 claims abstract description 7
- 239000013535 sea water Substances 0.000 claims description 11
- 241000237858 Gastropoda Species 0.000 claims description 2
- 238000007598 dipping method Methods 0.000 claims description 2
- 239000013505 freshwater Substances 0.000 claims description 2
- 238000002372 labelling Methods 0.000 claims 1
- 239000002245 particle Substances 0.000 claims 1
- 241001465754 Metazoa Species 0.000 abstract description 6
- 230000002045 lasting effect Effects 0.000 abstract description 3
- 230000000704 physical effect Effects 0.000 abstract 1
- 235000020639 clam Nutrition 0.000 description 19
- 241000543567 Rapana venosa Species 0.000 description 12
- 230000004083 survival effect Effects 0.000 description 10
- 241000237519 Bivalvia Species 0.000 description 9
- 241000620877 Ruditapes philippinarum Species 0.000 description 7
- 229920002101 Chitin Polymers 0.000 description 6
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 241000195493 Cryptophyta Species 0.000 description 3
- 238000005273 aeration Methods 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000008239 natural water Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000009394 selective breeding Methods 0.000 description 1
Classifications
-
- 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
- A01K61/50—Culture of aquatic animals of shellfish
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention relates to a shell Cal marking method applied to living bivalve shellfish, belonging to the technical field of aquatic animal marking, wherein the method comprises the steps of selecting shellfish offspring seeds with the shell length of 1-6mm, dissolving calcein in shellfish culture water to be marked, dip-dyeing the shellfish offspring seeds in the culture water dissolved with calcein for 72 hours, and normally feeding microalgae in the period; detecting whether the mark is successful by using blue light excitation light, and displaying a green mark to indicate that the mark of the shellfish is successful; in the marking process, the shellfish seedlings cannot be touched or moved, the physical effect on the shellfish is reduced to the minimum, the mark is kept lasting on the premise of not causing physiological damage to individual shellfish, and the pollution to the environment is avoided.
Description
Technical Field
The invention relates to a marking method of shellfish, in particular to a shell Cal marking method applied to living bivalve shellfish.
Background
The aquatic animal marking technology can be used for marking and releasing aquatic animals and investigating the resources and activity conditions of the aquatic animals, and can also be applied to large-scale selective breeding, such as family breeding and the like. In order to facilitate the distinction between different families and different treatment groups, the shellfish of different families and different treatment groups need to be marked. Common methods for marking common shellfish include tag marking and molecular marking, and the tag marking easily causes damage to the shellfish. The molecular marker is not easy to cause harm to animals, but the detection is complex and the cost is high. The general requirements of the marking technology are that the mark has little harm to animals, is simple and easy to implement, has low cost, is suitable for mass operation, has obvious mark, is not easy to fall off or disappear, and is easy to identify and detect.
Disclosure of Invention
In order to solve the technical problems, the invention provides an environmentally-friendly marking method which is obvious in marking, low in cost, simple in operation and free of physiological damage to the shellfish and is applied to the shell calcein (Cal) marking method of the living bivalve shellfish.
The invention is realized by the following technical scheme:
a shell Cal marking method applied to living bivalve shellfish is specifically as follows:
(1) selecting shellfish seeds to be marked, wherein the seed shells of the selected shellfish seeds have the length of 1-6 mm;
(2) dissolving calcein (Cal) in the shellfish culture water to be marked, wherein the concentration is 100 mg/L;
(3) dipping and dyeing the shellfish offspring seeds in the culture water with calcein dissolved in the step (2) for 72h, normally feeding microalgae, detecting whether the mark is successful by using blue light excitation light, and displaying a green mark to indicate that the shellfish mark is successful;
(4) the marked shellfish is normally fed, and then the mark is checked by blue light exciting light, and the green mark shows success except a newly grown part.
As a preferable scheme of the invention, in the step (3), the impregnation density of the shellfish seeds is 1000-4000 granules/square meter.
In a preferred embodiment of the present invention, in the step (3), the shellfish offspring seed is shellfish having a calcareous shell such as marine or freshwater bivalve, gastropod, etc.
Compared with the prior art, the invention has the beneficial effects that:
according to the marking method, 1-6mm of larvae are soaked in Cal (100mg/L) dissolved aquaculture water for 72 hours, and microalgae are normally fed in the period. Can ensure that the mark is kept lasting on the premise of not causing physiological damage to the shellfish individual and can not cause pollution to the environment.
Detailed Description
The technical solution of the present invention is further explained with reference to the following examples, but the scope of the present invention is not limited in any way by the examples.
Example 1
Taking a Ruditapes philippinarum mark with a shell length of 4-5mm as an example, the method sequentially comprises the following steps:
1) selecting 4-5mm Ruditapes philippinarum young seedling, and cleaning algae on the chitin surface of the young clam.
2) Dissolving Cal in pure seawater, selecting natural water body of shellfish natural living water area, preparing Cal (100mg/L) dip-dyeing solution, setting control group, and repeating the steps.
3) Ruditapes philippinarum larvae were dip-dyed for 72h at 2000 grains per square meter in 3L clear kegs, 30 grains each, with 2 normal feeds per day and normal aeration to simulate a flowing seawater environment, and the success of the mark was checked with a fluorophotometer (blue excitation light) after dip-dyeing. Under the bright field state, the chitin of the impregnated Ruditapes philippinarum has no obvious change.
4) The marked Ruditapes philippinarum was fed normally for two months, the water was changed all at once every two days, and then the mark was checked with a fluorophor scope (blue light excitation light).
The impregnated Ruditapes philippinarum larvae are normally fed for 69 days, water is completely changed every two days, the survival rate is recorded, and the marking conditions are detected one by using a fluorophor sight glass (blue light exciting light). After the clams are restored and maintained for 69 days, clear and bright green fluorescent marks can be generated by 100mg/L Cal-stained clams chitin. Neither the exhaust group nor the control group died in the 72h exhaust test. The clams are restored for 58 days, the average survival rate of the test group is 91 percent, and the survival rate of the clams is hardly influenced considering natural death. The clams are restored and maintained for 69 days, and the result of one-factor anova shows that the difference of the shell length of each experimental group and the shell length of the control group is not significant (P is more than 0.05). Thus, the fluorescent substance has no effect on the growth and survival of the clams.
Example 2
Taking a clam with a shell length of 2mm as an example, the method sequentially comprises the following steps:
1) selecting 1-2mm clam young seedlings, and cleaning algae on the surfaces of the clams.
2) Dissolving Cal in pure seawater, selecting natural water body of shellfish natural living water area, preparing Cal (100mg/L) dip-dyeing seawater with seawater, and setting control group.
3) Clam larvae were dip-dyed for 72h at 2000 grains per square meter in 3L clear kegs of 30 grains each, with 2 normal feeds per day and normal aeration to simulate a flowing seawater environment, and the success of the mark was checked with a phosphor scope (blue light excitation light) after dip-dyeing. Under the open field state, the impregnated clam chitin has no obvious change.
4) The marked clams were fed normally for 69 days with a total change of water every two days, after which the marking was checked with a fluorophor scope (blue excitation light).
The impregnated Ruditapes philippinarum larvae are normally fed for 69 days, water is completely changed at intervals of two days, the survival rate is recorded in the process, and the marking conditions are detected one by using a fluorophor sight glass (blue light exciting light). The clam is restored for 69 days, and clear and bright green fluorescent marks can be generated by 100mg/L Cal-stained clam chitin. Neither the exhaust group nor the control group died in the 72h exhaust test. The clam recovers and sustains for 58 days, and the average survival rate of the test group is more than 80 percent. The clam recovers and continues to be cultured for 69 days, and the result of single-factor anova shows that the difference of the shell length of each experimental group and the shell length of the control group is not significant (P is more than 0.05). Therefore, the fluorescent substance has no effect on the growth and survival of the clam.
Example 3
Taking a rapana venosa with the shell length of 1-2mm as an example, the method sequentially comprises the following steps:
1) selecting 1-2mm rapana venosa seedlings, and cleaning algae on the surface of rapana venosa.
2) Dissolving Cal in pure seawater, selecting natural water body of shellfish natural living water area, preparing Cal (100mg/L) dip-dyeing seawater with seawater, and setting control group.
3) Rapana venosa larvae were dip-dyed for 72h at 2000 grains per square meter in 3L clear kegs of 30 grains each, with 2 normal feeds per day and normal aeration to simulate a flowing seawater environment, and the success of the mark was checked with a phosphor-scope (blue-light excitation light) after dip-dyeing. The chitin of the rapana venosa has strong green fluorescent signals under the irradiation of blue light exciting light, and the dyed rapana venosa has no obvious change under the bright field state.
4) The marked rapana venosa was normally fed for two months with a total change of water every two days, after which the marking was checked with a fluoroscope (blue light excitation).
The impregnated rapana venosa larvae are normally fed for 69 days, water is completely changed every two days, the survival rate is recorded in the process, and the marking conditions are detected one by using a fluorophor sight glass (blue light exciting light). The young clams are restored and maintained for 69 days, and the clear and bright green fluorescent mark can be generated by the red rapana venosa dipped with 100mg/L Cal. Neither the exhaust group nor the control group died in the 72h exhaust test. The rapana venosa is recovered for 58 days, and the average survival rate of the test group is more than 80 percent. The rapana venosa is restored and continuously nourished for 69 days, and the result of one-factor anova shows that the difference of the shell length of each experimental group and the shell length of the control group is not significant (P is more than 0.05). Thus, the fluorescent substance had no effect on the growth and survival of the rapana venosa.
In conclusion, the marking method disclosed by the invention can provide a lasting fluorescent mark for the shells of the shellfish under the condition of not damaging the physiology of the shellfish seedlings, and has the advantages of clear mark, low cost and simple operation.
In practical applications, a person skilled in the art can reasonably select other parameters within the technical solution of the present invention, but the parameters are substantially the same as the technical solution protected by the present invention and still fall within the protection scope of the present invention.
Claims (3)
1. A shell Cal marking method applied to living bivalve shellfish is characterized by comprising the following steps:
(1) selecting shellfish seeds to be marked, wherein the seed shells of the selected shellfish seeds have the length of 1-6 mm;
(2) dissolving calcein in the shellfish culture water to be marked, wherein the concentration is 100 mg/L;
(3) dipping and dyeing the shellfish offspring seeds in the culture water with calcein dissolved in the step (2) for 72h, normally feeding microalgae, detecting whether the mark is successful by using blue light excitation light, and displaying a green mark to indicate that the shellfish mark is successful;
(4) the marked shellfish is normally fed, and then the mark is checked by blue light exciting light, and the green mark shows success except a newly grown part.
2. The method for shell Cal labeling of living bivalve shellfish as claimed in claim 1, wherein in said step (3), said shellfish seed dip-dyeing density is 1000-4000 particles/m.
3. A conch Cal marking method applied to living bivalve shellfish according to claim 1 characterized in that in said step (3) said shellfish offspring seed is shellfish with calcareous shells, such as seawater or freshwater bivalves, gastropods.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210231473.7A CN114868682A (en) | 2022-03-09 | 2022-03-09 | Shell Cal marking method applied to living bivalve shellfish |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210231473.7A CN114868682A (en) | 2022-03-09 | 2022-03-09 | Shell Cal marking method applied to living bivalve shellfish |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114868682A true CN114868682A (en) | 2022-08-09 |
Family
ID=82667882
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210231473.7A Withdrawn CN114868682A (en) | 2022-03-09 | 2022-03-09 | Shell Cal marking method applied to living bivalve shellfish |
Country Status (1)
| Country | Link |
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| CN (1) | CN114868682A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993016351A1 (en) * | 1992-02-17 | 1993-08-19 | Reson System A/S | System and method for testing and/or identification of objects including especially living fish-objects |
| JP2010213602A (en) * | 2009-03-16 | 2010-09-30 | Nagasaki Prefecture | Mixed feed for initial seedling of useful herbivorous shellfish, and method for feeding the same |
| CN102742532A (en) * | 2012-07-31 | 2012-10-24 | 中国海洋大学 | Cultivation method of novel strain of crassostrea gigas with pure purple left shells |
| CN106614216A (en) * | 2016-11-30 | 2017-05-10 | 中国水产科学研究院南海水产研究所 | Method for marking marine shellfishes by means of waterproof labels |
| CN107300546A (en) * | 2017-07-21 | 2017-10-27 | 浙江海洋大学 | The calcein labeling method and its detection means of Sepiella maindroni otolith |
| CN111727915A (en) * | 2020-06-30 | 2020-10-02 | 自然资源部第四海洋研究所(中国—东盟国家海洋科技联合研发中心) | Method for tracing heavy metal enrichment way of hanging-cultured bivalve shellfish |
-
2022
- 2022-03-09 CN CN202210231473.7A patent/CN114868682A/en not_active Withdrawn
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1993016351A1 (en) * | 1992-02-17 | 1993-08-19 | Reson System A/S | System and method for testing and/or identification of objects including especially living fish-objects |
| JP2010213602A (en) * | 2009-03-16 | 2010-09-30 | Nagasaki Prefecture | Mixed feed for initial seedling of useful herbivorous shellfish, and method for feeding the same |
| CN102742532A (en) * | 2012-07-31 | 2012-10-24 | 中国海洋大学 | Cultivation method of novel strain of crassostrea gigas with pure purple left shells |
| CN106614216A (en) * | 2016-11-30 | 2017-05-10 | 中国水产科学研究院南海水产研究所 | Method for marking marine shellfishes by means of waterproof labels |
| CN107300546A (en) * | 2017-07-21 | 2017-10-27 | 浙江海洋大学 | The calcein labeling method and its detection means of Sepiella maindroni otolith |
| CN111727915A (en) * | 2020-06-30 | 2020-10-02 | 自然资源部第四海洋研究所(中国—东盟国家海洋科技联合研发中心) | Method for tracing heavy metal enrichment way of hanging-cultured bivalve shellfish |
Non-Patent Citations (3)
| Title |
|---|
| 周珊珊 等: "贝类标志技术的研究进展", 《浙江海洋学院学报(自然科学版)》 * |
| 周珊珊: "魁蚶幼贝生境选择及标志技术研究", 《中国博士学位论文全文数据库 农业科技辑》 * |
| 耿倩 等: "荧光标记技术在增殖放流中的应用现状", 《水产科学》 * |
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Application publication date: 20220809 |
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| WW01 | Invention patent application withdrawn after publication |