CN103392632A - Half-smooth tonguefish fluorescence labeling method - Google Patents
Half-smooth tonguefish fluorescence labeling method Download PDFInfo
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- CN103392632A CN103392632A CN2013102521761A CN201310252176A CN103392632A CN 103392632 A CN103392632 A CN 103392632A CN 2013102521761 A CN2013102521761 A CN 2013102521761A CN 201310252176 A CN201310252176 A CN 201310252176A CN 103392632 A CN103392632 A CN 103392632A
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- 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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- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a half-smooth tonguefish fluorescence labeling method which comprises the steps of anesthesia and labeling. A composite fluorescence agent adopted in the method comprises, by weight, 30-50 parts of silica gel stuffing, 5-10 parts of cross-linking agent, 10-30 parts of colored fluorescence powder, and 0.1-1 part of cyclosporine A. The composite fluorescence agent further contains 0.01 microgram/milliliter-2 micrograms/milliliter of vitamin K and 0.5 microgram/milliliter-100 micrograms/milliliter of bile acid or glycocholic acid. The half-smooth tonguefish fluorescence labeling method is simple in operation, quick, low in cost and capable of meeting the requirement of labeling half-sooth tonguefish fry in batch, and fishes which are labeled are high in rejuvenation speed.
Description
Technical field
The present invention relates to a kind of releasing of aquatic animal labeling technique, relate in particular to a kind of method of fluorescence labeling Cynoglossus semilaevis.
Background technology
Cynoglossus semilaevis is one of Important Economic fingerling of CHINESE OFFSHORE bottom, and due to overfishing, resource reduces in a large number.Total man's work raising technology of Cynoglossus semilaevis breaks through at present, and in order to recover its fishery resources, coastal area has been carried out artificial seed's enhancement releasing successively., for better grasping the resource situation after Cynoglossus semilaevis is released, utilize the mark-technology of recapturing to carry out follow-up investigation to it.
Mark-the technology of recapturing is one of application technology in Fish behavior, biology, Ecological Basis research.It refers to and will catch in natural water area or the fish of artificial breeding is put back to natural water area after marking, can, according to relevant record and biological data, study the biological propertyes such as age, the growing state of its circuit that moves about, translational speed, distribution and this fingerling of growing, the characteristics of ingesting during recapture.
Present fish labeling technique both domestic and external mainly is divided into two large classes: external tracer method and inside and outside tracer method.External tracer method comprises listing mark, cuts the fin mark, the branding mark, and body internal labeling method comprises molecular labeling, coded wire tag, archive office's mark, biotelemetry mark, separate type satellite markers method etc.
Tongue sole build tongue sole side is flat, likes pasting ground swimming and moves ahead.Above-mentioned listing mark, not only larger to the injury of fish body, the notice plate during swimming on its fish body also can increase water resistance, affects its normal activity; The methods such as fin mark, branding mark of cutting are also larger to the injury of fish body, and are lower with the conservation rate of tense marker; Molecular labeling method, coded wire tag, archive office's mark, biotelemetry mark, separate type satellite markers etc. need to be purchased special marker instrument and fetch equipment, and be expensive, and complex operation.
Summary of the invention
A kind of method that the purpose of this invention is to provide fluorescence labeling Cynoglossus semilaevis, be intended to improve the mark Retention, makes that mark is more eye-catching is easy to identification, reduce simultaneously the injury to the mark fish, and the method method of operating is simple, and is with low cost.
The present invention is achieved in that a kind of method of fluorescence labeling Cynoglossus semilaevis, and it has following steps:
1. anaesthetize step: choose health, the Cynoglossus semilaevis that vigor is good, supported 1 to 2 day in first pond, not feeding; Then, Cynoglossus semilaevis being put into concentration is that the eugenol solution of 15mg/L～20mg/L soaks anesthesia; The anaesthetic treatment time is 1 minute～2 minutes; After soaking anesthesia in eugenol solution, fish is comparatively quiet, is convenient to marking operation, stops eating and can effectively improve survival rate.
2. markers step: the Cynoglossus semilaevis fish body after the anesthesia step process is fixed, and with syringe, composite fluorescence reagent is injected its epidermis and forms the fluorescence blob of viscose.Ciclosporin A has anti-rejection effect, can effectively prevent rejection, and tool improves and is marked as the effect of power.Silica filler and crosslinking agent can solidify the formation supporting construction, are convenient to visually observe and are convenient to take out.The discrimination of technical solution of the present invention is near 100%.In the situation that not by any identification facility, naked eyes can be seen color marker was significantly arranged daytime, had evening color marker can send light fluorescence; And under the ultraviolet lamp optical excitation, naked eyes can be seen obvious fluorescence.Adopt the vitamin K of this proportioning, can make experimental subjects have higher survival rate, and can prevent near the necrosis of organizing mark, improve the success rate of mark.The reagent of this proportioning is less to the degree of injury of fish body, does not affect the normal activity of Cynoglossus semilaevis after mark, and above the present invention of retention rate 98% of mark is simple to operate; fast; cost is low, can meet the needs of semi-smooth tongue sole offspring breed mark in batches, and the fish rejuvenation after mark is very fast.
The Optimized Measures of taking comprises: composite fluorescence reagent contains 30～50 parts of silica fillers, 5～10 parts of crosslinking agents, 10 to 30 parts of colored fluorescent powder, 0.1 to 1 part of Ciclosporin A by weight.Ciclosporin A has anti-rejection effect, can effectively prevent rejection, and tool improves and is marked as the effect of power.Silica filler and crosslinking agent can solidify the formation supporting construction, are convenient to visually observe and are convenient to take out.The discrimination of technical solution of the present invention is near 100%.In the situation that not by any identification facility, naked eyes can be seen color marker was significantly arranged daytime, had evening color marker can send light fluorescence; And under the ultraviolet lamp optical excitation, naked eyes can be seen obvious fluorescence.
Composite fluorescence reagent also contains the vitamin K of 0.01ug/ml～2ug ug/ml.Adopt the vitamin K of this proportioning, can make experimental subjects have higher survival rate, and can prevent near the necrosis of organizing mark, improve the success rate of mark.The reagent of this proportioning is less to the degree of injury of fish body, does not affect the normal activity of Cynoglossus semilaevis after mark, and the retention rate of mark is more than 98%.Simultaneously, experiment showed, that vitamin K antagonism rejection effect has certain synergy.
Composite fluorescence reagent also contains bile acid or glycocholic acid 0.5ug/ml～100ug/ml.The present invention is simple to operate, and fast, cost is low, can meet the needs of semi-smooth tongue sole offspring breed mark in batches.It also has the effect of wound repairing, and makes the fish rejuvenation after mark very fast.
The fluorescent glue block length is 5mm approximately, and width is 2mm approximately.In markers step, the syringe needle of syringe and Cynoglossus semilaevis fish body are 5 ° ~ 10 ° angles and thrust near the epidermis undertissue of fish body dorsal fin base portion, Yi Bian then extract slowly syringe needle, Yi Bian composite fluorescence reagent is injected epidermis undertissue.
The syringe needle of syringe was sterilized 10 ~ 15 minutes before use in 90 ~ 95% alcohol; After completing, markers step, with the potassium permanganate sea water solution that concentration is 20mg/L～50mg/L, fish body wound fluid is carried out soaking disinfection, soak time 1 ~ 2 minute, then the mark fish is put into clean sea water recover.
Compared with prior art, the invention has the advantages that:
L, owing to adopting the medical silica-gel agent that serves as a mark, less to the degree of injury of fish body, do not affect the normal activity of Cynoglossus semilaevis after mark, the retention rate of mark is more than 98%;
2, discrimination is near 100%.In the situation that do not borrow any identification facility, naked eyes can be seen color marker was significantly arranged daytime, had evening color marker can send light fluorescence; And under the ultraviolet lamp optical excitation, naked eyes can be seen obvious fluorescence;
3, the present invention is simple to operate, fast, can meet the needs of semi-smooth tongue sole offspring breed mark in batches;
4, tagged fluorescent agent cost of the present invention is lower, if disposable mark 10000 tail Cynoglossus semilaevis, the cost of every fish is very low.
Description of drawings
Fig. 1 is the operation photo of the embodiment of the present invention;
Fig. 2 is the effect photo of the embodiment of the present invention.
Embodiment
Below in conjunction with attached embodiment, the present invention is described in further detail.
Embodiment 1: a kind of method of fluorescence labeling Cynoglossus semilaevis, and it has following steps:
1. anaesthetize step: choose health, the Cynoglossus semilaevis that vigor is good, supported 2 days in first pond, not feeding; Then, Cynoglossus semilaevis being put into concentration is that the eugenol solution of 20mg/L soaks anesthesia; The anaesthetic treatment time is 2 minutes; After soaking anesthesia in eugenol solution, fish is comparatively quiet, is convenient to marking operation, stops eating and can effectively improve survival rate.
2. markers step: the Cynoglossus semilaevis fish body after the anesthesia step process is fixed, and with syringe, composite fluorescence reagent is injected its epidermis and forms the fluorescence blob of viscose.Ciclosporin A has anti-rejection effect, can effectively prevent rejection, and tool improves and is marked as the effect of power.Silica filler and crosslinking agent can solidify the formation supporting construction, are convenient to visually observe and are convenient to take out.The discrimination of technical solution of the present invention is near 100%.In the situation that not by any identification facility, naked eyes can be seen color marker was significantly arranged daytime, had evening color marker can send light fluorescence; And under the ultraviolet lamp optical excitation, naked eyes can be seen obvious fluorescence.Adopt the vitamin K of this proportioning, can make experimental subjects have higher survival rate, and can prevent near the necrosis of organizing mark, improve the success rate of mark.The reagent of this proportioning is less to the degree of injury of fish body, does not affect the normal activity of Cynoglossus semilaevis after mark, and above the present invention of retention rate 98% of mark is simple to operate; fast; cost is low, can meet the needs of semi-smooth tongue sole offspring breed mark in batches, and the fish rejuvenation after mark is very fast.
The Optimized Measures of taking comprises: composite fluorescence reagent contains 30ug silica filler, 10ug crosslinking agent, 30 ug colored fluorescent powder, 0.1 ug Ciclosporin A by weight.Approximately the colloidal solution of 1ml is stand-by to make volume.Marks as a large amount of in needs, double to enlarge on this basis, and formula expansion mode hereinafter repeats no more.Ciclosporin A has anti-rejection effect, can effectively prevent rejection, and tool improves and is marked as the effect of power.Silica filler and crosslinking agent can solidify the formation supporting construction, are convenient to visually observe and are convenient to take out.The discrimination of technical solution of the present invention is near 100%.In the situation that not by any identification facility, naked eyes can be seen color marker was significantly arranged daytime, had evening color marker can send light fluorescence; And under the ultraviolet lamp optical excitation, naked eyes can be seen obvious fluorescence.
Every milliliter of composite fluorescence reagent also contains the vitamin K of 0.01ug.Adopt the vitamin K of this proportioning, can make experimental subjects have higher survival rate, and can prevent near the necrosis of organizing mark, improve the success rate of mark.The reagent of this proportioning is less to the degree of injury of fish body, does not affect the normal activity of Cynoglossus semilaevis after mark, and the retention rate of mark is more than 98%.
Every milliliter of composite fluorescence reagent also contains bile acid or glycocholic acid 50ug.The present invention is simple to operate, and fast, cost is low, can meet the needs of semi-smooth tongue sole offspring breed mark in batches, and the fish rejuvenation after mark is very fast.
The fluorescent glue block length is 5mm approximately, and width is 2mm approximately.In markers step, the syringe needle of syringe and Cynoglossus semilaevis fish body are 5 ° ~ 10 ° angles and thrust near the epidermis undertissue of fish body dorsal fin base portion, Yi Bian then extract slowly syringe needle, Yi Bian composite fluorescence reagent is injected epidermis undertissue.
The syringe needle of syringe was sterilized 10 ~ 15 minutes before use in 90 ~ 95% alcohol; After completing, markers step, with the potassium permanganate sea water solution that concentration is 20mg/L～50mg/L, fish body wound fluid is carried out soaking disinfection, soak time 1 ~ 2 minute, then the mark fish is put into clean sea water recover.
Embodiment 2: test procedure is as follows:
1, the preparation of tagged fluorescent agent: adopt bicomponent normal temperature medical grade silicon rubber (model is SY-1, by The Fourth Military Medical University's development), colored fluorescent powder and Ciclosporin A, medical grade silicon rubber is comprised of silica filler and crosslinking agent.It is characterized in that tagged fluorescent agent is that silica filler, crosslinking agent, colored fluorescent powder and Ciclosporin A evenly mix by the weight ratio of 50:10:30:1 the coloured silica gel one fluorescence thickness colloid that is mixed with.To configure tagged fluorescent agent and inject in the 1ml injector for medical purpose, the needle gauge of syringe is selected diameter 0. 45mm.
Above-mentioned crosslinking agent composition is two propylene triamines [DPTA, molecular formula is H2N(CH2) 3 NH(CH2) 3NH2]; Colored fluorescent powder is that Mitsubishi chemical company produces, minute three kinds of colors: salmon pink fluorescent material (model BR-102F), green emitting phosphor (model BG-201B) and yellow fluorescent powder (model BY-201A), can select as required suitable color; The Ciclosporin A molecular formula is C
62H
111N
11O
12, to be produced by Zhongmei Huadong Pharmaceutical Co., Ltd. Hangzhou, Main Function is for increasing the anti-rejection of fish body.
2, the anesthesia of mark fish: choose health, Cynoglossus semilaevis 200 tails that vigor is good, the long 8cm that surpasses of body; Cynoglossus semilaevis must be supported 1-2 days, not feeding in first pond; Cynoglossus semilaevis is put into eugenol solution that concentration is 15～20mg/L etc. soak anesthesia, anesthesia duration is 1 ~ 2 minute.
3, fish body tag: postanesthetic Cynoglossus semilaevis lies on the workbench that is covered with wet towel, a hand is worn wet yarn gloves, gently the fish body is fixed, the syringe needle of above-mentioned small-sized syringe and fish body are 5 ° ~ 10 ° angles thrust near fish body dorsal fin base portion epidermis undertissue, then extract slowly syringe needle on one side, on one side tagged fluorescent agent is injected epidermis, make it formation and be about 5mm, coloured fluorescence blob of viscose of wide approximately 2mm, see accompanying drawing 1.
The syringe needle of syringe was sterilized 10 ~ 15 minutes before use in 90 ~ 95% alcohol; With concentration, being that the potassium permanganate seawater of 50mg/L is molten after mark carries out soaking disinfection to fish body wound fluid, soak time 1 ~ 2 minute, then the mark fish is put into clean sea water recover.
200 tail tag note fishes are supported temporarily in Zhoushan Landtek marine organisms research institute fishpond and observe, find after 10 days that all are remarkable by Tagging fish sign effect, the marking fluid of fish surface is high-visible, sees accompanying drawing 2.Continue to observe through the fish of mark, mark after 1 month survival rate reach 99%, the mark discrimination reaches 100%.
Although in conjunction with preferred embodiment, described the present invention; so it is not in order to limit the present invention; any those skilled in the art; without departing from the spirit and scope of the present invention; can implement to the theme of listing displacement and the modification of various changes, coordinate here, so protection scope of the present invention is as the criterion when looking the claim restricted portion that proposes.
Claims (7)
1. the method for a fluorescence labeling Cynoglossus semilaevis is characterized in that having following steps:
1. anaesthetize step: choose health, the Cynoglossus semilaevis that vigor is good, supported 1 to 2 day in first pond, not feeding; Then, described Cynoglossus semilaevis being put into concentration is that the eugenol solution of 15mg/L～20mg/L soaks anesthesia; The described anaesthetic treatment time is 1 minute～2 minutes;
2. markers step: the Cynoglossus semilaevis fish body after the anesthesia step process is fixed, and with syringe, described composite fluorescence reagent is injected its epidermis and forms the fluorescence blob of viscose.
2. the method for a kind of fluorescence labeling Cynoglossus semilaevis according to claim 1, it is characterized in that: described composite fluorescence reagent contains 30～50 parts of silica fillers, 5～10 parts of crosslinking agents, 10 to 30 parts of colored fluorescent powder, 0.1 to 1 part of Ciclosporin A by weight; Described crosslinking agent is two propylene triamines.
3. the method for a kind of fluorescence labeling Cynoglossus semilaevis according to claim 2, it is characterized in that: described composite fluorescence reagent also contains the vitamin K of 0.01ug/ml～2ug ug/ml.
4. the method for according to claim 2 or 3 described a kind of fluorescence labeling Cynoglossus semilaevis, it is characterized in that: described composite fluorescence reagent also contains bile acid or glycocholic acid 0.5ug/ml～100ug/ml.
5. the method for a kind of fluorescence labeling Cynoglossus semilaevis according to claim 1, it is characterized in that: described fluorescent glue block length is 5mm approximately, and width is 2mm approximately.
6. the method for a kind of fluorescence labeling Cynoglossus semilaevis according to claim 1, it is characterized in that: in markers step, the syringe needle of described syringe and described Cynoglossus semilaevis fish body are 5 ° ~ 10 ° angles and thrust near the epidermis undertissue of fish body dorsal fin base portion, then extract slowly syringe needle on one side, on one side composite fluorescence reagent is injected epidermis undertissue.
7. the method for a kind of fluorescence labeling Cynoglossus semilaevis according to claim 1, it is characterized in that: the syringe needle of described syringe was sterilized 10 ~ 15 minutes before use in 90 ~ 95% alcohol; After completing, markers step, with the potassium permanganate sea water solution that concentration is 20mg/L～50mg/L, fish body wound fluid is carried out soaking disinfection, soak time 1 ~ 2 minute, then the mark fish is put into clean sea water recover.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310252176.1A CN103392632B (en) | 2013-06-24 | 2013-06-24 | Half-smooth tonguefish fluorescence labeling method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310252176.1A CN103392632B (en) | 2013-06-24 | 2013-06-24 | Half-smooth tonguefish fluorescence labeling method |
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| CN103392632A true CN103392632A (en) | 2013-11-20 |
| CN103392632B CN103392632B (en) | 2015-04-29 |
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| CN111011286A (en) * | 2019-12-02 | 2020-04-17 | 中国水产科学研究院黑龙江水产研究所 | Acipenser schrenki and huso dauricus juvenile individual marking method |
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| CN113080111A (en) * | 2021-04-15 | 2021-07-09 | 山东龙昌动物保健食品有限公司 | Application of bile acid in improving intestinal health and cultivation survival rate of cynoglossus semilaevis |
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