CN102273418A - Fluorescent labeling method of fish otolith - Google Patents
Fluorescent labeling method of fish otolith Download PDFInfo
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- CN102273418A CN102273418A CN2011101727109A CN201110172710A CN102273418A CN 102273418 A CN102273418 A CN 102273418A CN 2011101727109 A CN2011101727109 A CN 2011101727109A CN 201110172710 A CN201110172710 A CN 201110172710A CN 102273418 A CN102273418 A CN 102273418A
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Abstract
The invention discloses a novel fluorescent labeling method of fish otolith. According to the invention, the method comprises the steps of mixing an alizarin complex and a fish compound feed; making a mixture into a labeled feed; and then feeding a fish to be labeled with the labeled feed so that an identifiable fluorescent label can be detected on the labeled fish otolith. By using the novel fluorescent labeling method of the fish otolith disclosed by the invention, the defects of larger damage to fish body, high label loss ratio, high labeling price, low retention rate, limited label number and the like in other labeling methods are avoided. By using the method, waste and pollution problems of an alizarin complex indicator in a method for fluorescently labeling the fish otolith through dipping are solved; and the method has the advantages of simplicity in operation, low cost and easiness for promotion and is suitable for labeling the fish otolithes with different sizes and types, in particular for large-scale launching labeling.
Description
Technical field
The present invention relates to the fluorescently-labeled method of a kind of fish otolith.
Background technology
The otolith of fish is a kind of calcified tissue, is present in the labyrinthus membranaceus of inner ear, plays the tissue of the balance and the sense of hearing, and bony fish generally has sagitta, little otolith and 3 pairs of otoliths of star otolith, is present in respectively in sacculus, utricle and the lagena of inner ear.Fish are carrying out in the process of mass exchange with external environment, and the fish body that constantly is ingested of the chemical element in the environment through a series of metabolism, circulation, finally can deposit on the otolith in the fish body.Therefore, can make fish on otolith, form specific markers, to be different from the nature population by changing the environment of fish growth.
The fish multiplication recruitment evaluation of releasing adopts the sign method of recapturing usually, and this just need carry out mark to fish and distinguish release colony and wild population before releasing.Labeling method at present commonly used list four kinds of tracer methods, external tracer method, body internal labeling method, blotelemetry, wherein external tracer method comprises the fin method of cutting, branding method, pigment injection etc., but body internal labeling method comprises coding metal mark, passive integration radar mark, built-in sighting target and chemical labeling method.
The otolith fluorescence labeling belongs to a kind of of chemical labeling method, selects fluorescent dye usually for use, as alizarin complex compound and calcein etc., because they can combine with calcium, and under the deposition, can detect fluorescence such as redness, green under fluorescence microscope in otolith.This method has avoided other labeling method bigger to the injury of fish body, mark loss rate height, mark costs an arm and a leg, shortcomings such as conservation rate is low, mark limited amount, and this method is applicable to the fish colony of different sizes, has the labeling effciency height, advantages such as cost is low, and the mark retention time is long are the desirable means of extensive fry mark, the work such as recruitment evaluation of releasing for artificial releasing population identification and propagation provides new approaches, has application promise in clinical practice.
At present, the otolith fluorescence labeling adopts infusion method usually, promptly fluorescent dye is dissolved in the water, and puts into then and takes out after fry is raised a period of time, will form the fluorescence labeling ring on the fish otolith.The most frequently used fluorescent marker is alizarin complex compound and quadracycline.But studies show that, quadracycline solution has bigger toxicity to the fish body, and in various degree incompatible or dead can appear in fish when concentration was high, and the mark effect is relatively poor, therefore general not as the fluorescent marker of recommending, and the alizarin complex compound is proper fluorescent marker.Existing research report shows: could form mark wheel comparatively clearly after generally soaking with the above alizarin complex solution of 100mg/L on the fish otolith.How woods etc. is " soaking heavy mouthful of schizothoracin prelarva otolith of mark with fluorescent material " spring, 2008 the 3rd phases of Sichuan animal, pointed among the p331-314, soaking mark with fluorescent material counterweight mouth schizothoracin prelarva shows, the alizarin complex solution of 200～300mg/L soaks 12h counterweight mouth schizothoracin prelarva otolith good mark effect, all can detect tangible Marking ring under fluorescence and the visible light; The mark of 100～150mg/L can detect the fluorescence labeling ring, but Marking ring is fainter under the visible light.Identical soak time, along with the increase of soaking concentration, Marking ring intensity increases on the otolith; Identical soaking concentration, along with the increase of soak time, Marking ring intensity increases on the otolith.In three pairs of otoliths, little otolith and sagitta are responsive to the alizarin complex compound, and star otolith susceptibility is lower.The mark poor effect of quadracycline when soaking concentration is 50～160mg/L, all can not detect Marking ring on three pairs of otoliths, and the quadracycline soak of 110～160mg/L has higher causing to lose consciousness or lethal effect to prelarva simultaneously.Therefore, the alizarin complex compound is that counterweight mouth schizothoracin fry carries out the proper fluorescent material of chemical labeling, and quadracycline is not suitable for the otolith of this fish of mark.And for example pay from east etc. pointedly " soaking mark mullet son, postlarva otolith with fluorescent material ", the alizarin complex solution of 100～200mg/L soaks 24h has good mark effect to mullet son, postlarva otolith.
But according to physical property as can be known the alizarin complex compound be slightly soluble in water, but need to use other cosolvents just can reach the concentration of mark.And the use of cosolvent has certain side effect to fish.Therefore, use the method for soaking the mark otolith, have only the label of minute quantity can be deposited on the otolith, most of label or be present in the water, can't obtain utilizing, the availability of label is extremely low, exists alizarin complex compound usage amount big, waste problems such as serious, and can cause certain harmful effect environment.
Summary of the invention
The purpose of this invention is to provide a kind of novel fluorescently-labeled method of fish otolith.
The technical solution used in the present invention is:
The fluorescently-labeled method of a kind of fish otolith may further comprise the steps:
1) alizarin complex compound and fish meal are mixed, make and contain the alizarin complex compound in the feed, obtain the mark feed;
2) usage flag forage feed fish to be marked.
Preferably, the time of usage flag forage feed fish to be marked is not shorter than 24 hours.
Preferably, the content of alizarin complex compound accounts for 2%～10% of mark feed gross mass.
The invention has the beneficial effects as follows:
The inventive method avoided listing tracer method and external tracer method to the injury of fish body the big and high problem of mark loss rate, wait other body internal labeling method to compare with coding metal mark with passive integration radar mark, cheap and the suitable extensive mark of mark carries out the otolith fluorescence labeling with traditional infusion method and compares and more save the alizarin complex compound.The present invention can be according to fingerling class to be marked, size and quantity, the amount and the time of the mark feed of determining to throw something and feed, be a kind of can blanket labeling method.
In the inventive method, the alizarin complex compound is ingested in the fish body, does not almost have label to be dispersed in the water body, has improved the availability of label greatly.This has also reduced the potentially contaminated of label to water body simultaneously.
Description of drawings
Fig. 1 be Guangdong triangular bream sagitta green-yellow light, blue light and purple light excited under fluoroscopic image;
After Fig. 2 is to use high alizarin complex compound content feed (1:10) to feed, the little otolith of Guangdong triangular bream green-yellow light, blue light and purple light excited under fluoroscopic image;
After Fig. 3 is to use low alizarin complex compound content feed (1:40) to feed, the little otolith of Guangdong triangular bream green-yellow light, blue light and purple light excited under fluoroscopic image.
Embodiment
The fluorescently-labeled method of a kind of fish otolith may further comprise the steps:
1) alizarin complex compound and fish meal are mixed, make and contain the alizarin complex compound in the feed, obtain the mark feed;
2) usage flag forage feed fish to be marked.
Preferably, the time of usage flag forage feed fish to be marked is not shorter than 24 hours.If the content height of alizarin complex compound in the mark feed can suitably shorten the nursing time, otherwise, but proper extension.
Preferably, the content of alizarin complex compound accounts for 2%～10% of mark feed gross mass, is about to ratio that alizarin complex compound and manufactured feed be about 1:10～1:40 according to mass ratio and mixes and get final product.
Below in conjunction with embodiment, further specify the present invention.
Embodiment 1
1) selects for use the alizarin complex compound as fluorescent marker, it is mixed according to mass ratio 1:10 with compound feed used for fish, add low amounts of water, wetting feed and alizarin complex compound stir, and make whole feeds be aubergine, dry, grind the mark feed of making suitable particle size;
2) throw something and feed sufficient mark forage feed Guangdong triangular bream juvenile fish more than 24 hours.
Mark finishes, and takes out the otolith of Guangdong triangular bream juvenile fish, cleans otolith with absolute ethyl alcohol, removes the otolith surface impurity, observes otolith under fluorescence microscope, promptly as seen takes turns as the otolith mark of Fig. 1 and Fig. 2.Wherein: 1 is green-yellow light excites down the observation image; 2 are the blue-light excited image of observing down; 3 are the purple light excited image of observing down.
1) the alizarin complex compound is mixed in the ratio of 1:40 with fish meal, make the mark feed;
2) throw something and feed sufficient mark forage feed Guangdong triangular bream juvenile fish more than 24 hours.
Mark finishes, and takes out the otolith of Guangdong triangular bream juvenile fish, cleans otolith with absolute ethyl alcohol, removes the otolith surface impurity, observes otolith under fluorescence microscope, finds to be formed with on little otolith mark wheel clearly.Its result as shown in Figure 3.Wherein: 1 observes image for green-yellow light excites down; 2 are the blue-light excited image of observing down; 3 are the purple light excited image of observing down.
The mark feed
The mark feed contains the alizarin complex compound.Preferably, for guaranteeing normally searching for food of fish, the alizarin complex compound accounts for about 2～10% of feed.Can be by with alizarin complex compound and common fish meal, mix as the extensive stock feed and to obtain.
The ratio of alizarin complex compound and compound feed used for fish can be adjusted accordingly according to kind and the difference in size thereof of fish to be marked.The mark feed that is used for the fish of easy mark, the addition of alizarin complex compound can be low slightly; Want the fish of mark big more, the addition of alizarin complex compound can be high slightly.
For obtaining good mark effect, preferably much want 24 hours with the time of mark forage feed fish to be marked, for obtaining more significantly mark wheel, can prolong time with the mark forage feed.
Claims (3)
1. fluorescently-labeled method of fish otolith may further comprise the steps:
1) alizarin complex compound and fish meal are mixed, make and contain the alizarin complex compound in the feed, obtain the mark feed;
2) usage flag forage feed fish to be marked.
2. the fluorescently-labeled method of a kind of fish otolith according to claim 1, it is characterized in that: the time of usage flag forage feed fish to be marked is not shorter than 24 hours.
3. the fluorescently-labeled method of a kind of fish otolith according to claim 1, it is characterized in that: the content of alizarin complex compound accounts for 2%～10% of mark feed gross mass.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960274A (en) * | 2012-11-20 | 2013-03-13 | 中国长江三峡集团公司 | Method for marking fish otolith by fluorescent substances |
| CN103181352A (en) * | 2013-04-10 | 2013-07-03 | 中国水产科学研究院长江水产研究所 | Fish branchiostegite marking method |
| CN103875573A (en) * | 2014-04-11 | 2014-06-25 | 浙江省海洋水产研究所 | Method for marking juvenile sciaenidae |
| CN104106481A (en) * | 2014-06-23 | 2014-10-22 | 中国水产科学研究院东海水产研究所 | Marking method for proliferation and releasing of large yellow croakers |
| 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 |
| 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 |
| CN112852408A (en) * | 2020-12-30 | 2021-05-28 | 西藏自治区农牧科学院水产科学研究所 | Labeling reagent and labeling method for lasalosijiri fish |
| CN115281140A (en) * | 2022-08-12 | 2022-11-04 | 复旦大学 | Aquatic product biological marker culture method |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102960274A (en) * | 2012-11-20 | 2013-03-13 | 中国长江三峡集团公司 | Method for marking fish otolith by fluorescent substances |
| CN103181352A (en) * | 2013-04-10 | 2013-07-03 | 中国水产科学研究院长江水产研究所 | Fish branchiostegite marking method |
| CN103181352B (en) * | 2013-04-10 | 2015-03-04 | 中国水产科学研究院长江水产研究所 | Fish branchiostegite marking method |
| CN103875573A (en) * | 2014-04-11 | 2014-06-25 | 浙江省海洋水产研究所 | Method for marking juvenile sciaenidae |
| CN103875573B (en) * | 2014-04-11 | 2016-05-25 | 浙江省海洋水产研究所 | A kind of method that indicates Shishou section juvenile fish |
| CN104106481A (en) * | 2014-06-23 | 2014-10-22 | 中国水产科学研究院东海水产研究所 | Marking method for proliferation and releasing of large yellow croakers |
| CN104106481B (en) * | 2014-06-23 | 2016-03-02 | 中国水产科学研究院东海水产研究所 | A kind of method of large yellow Crocker enhancement releasing mark |
| 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 |
| 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 |
| CN112852408A (en) * | 2020-12-30 | 2021-05-28 | 西藏自治区农牧科学院水产科学研究所 | Labeling reagent and labeling method for lasalosijiri fish |
| CN112852408B (en) * | 2020-12-30 | 2024-01-23 | 西藏自治区农牧科学院水产科学研究所 | Marking reagent and marking method for Lassa nojirimi |
| CN115281140A (en) * | 2022-08-12 | 2022-11-04 | 复旦大学 | Aquatic product biological marker culture method |
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