Fluorescence labeling method for sea cucumber larvas
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A technology for fluorescent labeling and larvae, which can be used in fish farming, application, climate change adaptation, etc., and can solve problems such as prone to inflammatory reactions and inability to achieve effective labeling
Inactive Publication Date: 2011-08-31
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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The body of sea cucumbers contains high levels of autolytic enzymes. Domestic and foreign studies have shown that traditional physical markers, such as punctures on the body surface and
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Embodiment 1
[0017] Get the rough sea cucumber larvae whose individual average weight is 4.50g, and take the soaking mode to mark, and the specific implementation method is as follows:
[0018] 1. Prepare calcein (Calcein) with sterilized water to make a mother solution with a concentration of 200mg / ml, and store it in the dark at 4°C.
[0019] 2. Collect the sea cucumber larvae to be marked, put them in a 50L opaque open basin, add 10L of seawater, and inflate continuously to make the dissolved oxygen in the water reach 5.0mg / L.
[0020] 3. Add 10ml of calcein mother liquor to the seawater in the open basin to make the final concentration 200mg / L, stir gently to make the mother liquor evenly distributed.
[0021] 4. Shade the open basin with a shading cloth, and keep inflating continuously to make the dissolved oxygen in the water reach 5.0mg / L, and leave it for 22 hours.
[0022] 5. After standing for 22 hours, add 10L of fresh seawater to the basin, add once every 8 hours, and add 3 ti...
Embodiment 2
[0027] Take sea cucumber larvae with an average individual weight of 15.40g and mark them by injection. The specific implementation method is as follows:
[0028] 1. Prepare calcein (Calcein) with sterilized water to make a mother solution with a concentration of 200mg / ml, and store it in the dark at 4°C.
[0029] 2. Collect the sea cucumber larvae to be marked, put them in a 50L opaque open basin, add 40L of seawater, and inflate continuously to make the dissolved oxygen in the water reach 5.0mg / L.
[0030] 3. In a dark environment, take 0.1ml of calcein mother solution in a clean beaker, dilute the mother solution with 99.9ml of sterilized seawater to make the final concentration 200mg / L, and store in the dark.
[0031] 4. Take the diluted calcein diluted solution with a 2ml sterile syringe, and inject it into the body cavity of the sea cucumber at a dose of 0.3ml per tail, and the injection site is the side of the rear third of the sea cucumber body.
[0032] 5. Shade the ...
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Abstract
The invention provides a fluorescence labeling method for sea cucumber larvas. Sea cucumber is one of rare marine products, and wild sea cucumber resources are increasingly deficient. Artificial propagation and releasing is an important method for restoring the wild sea cucumber resources. In order to assess propagation and releasing effects, released individuals need to be regularly captured back, so that how to effectively distinguish artificially propagated individuals from wild individuals and establish a labeling method becomes an urgent need in the industry. A lot of sclerites are distributed at the body wall of the sea cucumber, and a main ingredient of the sclerite is calcium carbonate. Aiming at the characteristic, the invention provides a fluorescence labeling method for sea cucumber larvas, which comprises the following steps: soaking the sea cucumber larvas for 20-24 hours in a dark environment by using 200 mg/L calcein, or injecting the 200 mg/L calcein into sea cucumber coelomic cavities for labeling through an injection way. After labeling, more than 50% of the sclerites in the body wall are infected with the calcein, and fluorescence detection rate of the sclerites labeling the individuals is still above 5% after breeding for a year. The fluorescence labeling method can not affect the ingestion and growth of the sea cucumber larvas, and the marker calcein can be thoroughly removed out of the body through metabolism of the sea cucumber two years later.
Description
technical field [0001] The invention belongs to the field of marking methods for marine organisms, in particular to a fluorescent marking method for sea cucumber larvae. Background technique [0002] Sea cucumber is one of the precious seafood, and wild sea cucumber resources are increasingly scarce. Releasing artificially is an important method to restore wild sea cucumber resources. In order to evaluate the effect of multiplication and release, it is necessary to capture and release individuals regularly, so it is necessary to effectively distinguish artificially multiplied individuals from wild individuals. The body of sea cucumbers contains high levels of autolytic enzymes. Domestic and foreign studies have shown that traditional physical markers, such as punctures on the body surface and subcutaneous implantation, are prone to inflammatory reactions at the puncture or implantation sites, and the attachments are quickly removed. less than the purpose of effective marki...
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IPC IPC(8): A01K61/00
CPCY02A40/81
Inventor 张吕平胡超群夏建军赵哲
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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