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Calcein labeling method and detection method of Manson's needle-free cuttlefish otolith

A technology of needleless cuttlefish and calcein, which is applied in the direction of material analysis, measurement devices, and test sample preparation through optical means, and can solve problems such as inability to achieve release marking, damage to cuttlefish larvae, and difficulty in injection marking , to achieve the effect of convenient observation and comparison, simple marking method and quick operation

Active Publication Date: 2020-01-07
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the cuttlefish larvae are very small, and injection marking is very difficult, which will damage the cuttlefish larvae and affect their survival rate, and large-scale release and marking cannot be achieved.

Method used

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  • Calcein labeling method and detection method of Manson's needle-free cuttlefish otolith
  • Calcein labeling method and detection method of Manson's needle-free cuttlefish otolith

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] The calcein labeling method of man's needleless cuttlefish otolith comprises the following steps:

[0016] 1) Pretreatment of aquaculture water: Put the purified seawater into the slow-release labeled aquaculture box and slightly inflate it as the aquaculture water body, then add EDTA to the aquaculture water body, the amount of EDTA added per ton of aquaculture water is 6g, fully dissolved and then used , the EDTA in this step can undergo a complex reaction with free calcium ions in seawater to form a stable water-soluble complex, avoiding the loss of calcein used in the subsequent labeling step due to interfering ions, ensuring a stable labeling concentration and The stability of the marking effect;

[0017] 2) Preparation of marking soaking solution: add calcein to the breeding water to make a solution with a concentration of 300mg / L, expose to the sun for 12 hours, then add Miserin, the weight ratio of Calcein to Miserin is 1:0.25, mix well , that is, the marked so...

Embodiment 2

[0024] The calcein labeling method of man's needleless cuttlefish otolith comprises the following steps:

[0025] 1) Put the purified seawater into the slow-release marked culture box and slightly inflate it as the culture water body, then add EDTA to the culture water body, the amount of EDTA added per ton of culture water is 5g, fully dissolved and then set aside, in this step EDTA can undergo a complex reaction with free calcium ions in seawater to form a stable water-soluble complex, avoiding the loss of calcein used in the subsequent labeling step due to interfering ions, and ensuring stable labeling concentration and labeling effect;

[0026]2) Add calcein to the breeding water to make a solution with a concentration of 350mg / L, expose it to the sun for 13 hours, then add Miserin, the weight ratio of Calcein to Miserin is 1:0.3, mix well, and the mark is soaked In this step, Miserin can form a mixed complex with high-intensity fluorescence with calcein and calcified tiss...

Embodiment 3

[0030] 1. Marking effect:

[0031] Test group is embodiment 1; Blank group is not marked, and all the other steps are exactly the same as embodiment 1; The labeling of control group adopts the calcein solution that concentration is 300mg / L, does not add Miserin, all the other steps and embodiment 1 are complete same. The clarity of fluorescent markers is graded on a scale of 0–5, with grade 0: no markers visible under a fluorescent microscope; grade 1: faint markers visible under a fluorescent microscope; grade 2: markers easily discernible under a fluorescent microscope; Grade 3: Bright marks can be seen under a fluorescent microscope; Grade 4: Marks can be seen under natural transmitted light; Grade 5: Clear marks can be seen under natural transmitted light. If the marking level is ≥2, it can be considered as a good marking effect that can be detected. Eight otoliths of squid mansoni were selected in each group for detection, and the average value was taken. The test resul...

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Abstract

The invention discloses a marking method for calcein of sepiella maindroni otolith and a detection method thereof. The marking method comprises the following steps: adding calcein into aquaculture water, thereby preparing into a solution in concentration of 200-400mg / L; exposing to the sun; adding mianserin and uniformly mixing, thereby acquiring a marked soaking solution; hungering the to-be-marked sepiella maindroni and then soaking in the marked soaking solution; and soaking in fresh seawater after ending the soaking in the marked soaking solution, thereby acquiring the mark of the sepiella maindroni otolith. The invention has the beneficial effects that the marking method disclosed by the invention is simple, the purpose of nondestructive marking can be achieved and the large-scale release marking action can be realized at one time; the mixed compound with high-strength fluorescent light can be formed according to the marking method, the result observation and comparison become convenient and the marking time is long-lasting; the marking method is suitable for the sepiella maindroni in different stages; and an effective method is supplied for evaluating the proliferation release effect of the sepiella maindroni.

Description

technical field [0001] The invention relates to a labeling method for aquatic animals, in particular to a calcein labeling method and a detection means for manson's needle-free cuttlefish otolith. Background technique [0002] Sepiella japonica was once one of the four major fisheries in my country's fishing grounds. It has strong adaptability to temperature and is widely distributed in my country's coastal areas. It is a traditional fishery species with high economic value. Since the 1970s, a large number of juvenile squid and overwintering squid have been caught by various fishing gears such as trawl nets and stretch nets, resulting in the transition from growth-type overfishing to supplementary-type overfishing, which has destroyed the ecological balance of fishery resources, resulting in The resources of Mansoni squid in Zhejiang fishing grounds are almost exhausted. Since 2006, large-scale breeding and release of fertilized eggs and larvae of Squid mansoni have been ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64G01N1/28G01N1/30A01K61/00
CPCA01K61/00G01N1/2813G01N1/286G01N1/30G01N21/6402G01N21/6428G01N21/6458G01N2001/2866G01N2021/6441Y02A40/81
Inventor 龚理吕振明窦灿丰刘立芹王天明杨静文吴常文
Owner ZHEJIANG OCEAN UNIV
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