CN103231285B - A kind of preparation method of mandarin fish otolith cross section abrasive disc and application thereof - Google Patents
A kind of preparation method of mandarin fish otolith cross section abrasive disc and application thereof Download PDFInfo
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- CN103231285B CN103231285B CN201310147893.8A CN201310147893A CN103231285B CN 103231285 B CN103231285 B CN 103231285B CN 201310147893 A CN201310147893 A CN 201310147893A CN 103231285 B CN103231285 B CN 103231285B
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Abstract
The invention discloses a kind of preparation method and application of mandarin fish otolith cross section abrasive disc, belong to ecology of fishes field.The present invention chooses sagitta as research material, abrasive disc is carried out to its cross section, the otolith abrasive disc made is put under the microscope, under incident light irradiates, also measure the width between wheel line by the CCD software qualification age, observe the wheel line distribution characteristics of otolith abrasive disc, the age of qualification mandarin fish.Compared with conventional identification method, the method achieves takes turns the clear visual of line to mandarin fish otolith, and the side of otolith core deflection otolith abrasive disc, forms linear nucleus district; From otolith core, the opposite side towards otolith forms the zone that bright band and blanking bar interlock, and bright band is wider, formation time be spring to autumn, blanking bar is narrower, is formed at winter, is annual ring.The wheel line that this light and shade replaces, makes the age more easily distinguish and interpretation, thus substantially increases the accuracy of mandarin fish Age Determination.
Description
Technical field
The invention belongs to ecology of fishes field, be specifically related to a kind of preparation method of mandarin fish otolith cross section abrasive disc, also relate to the application of a kind of mandarin fish otolith cross section abrasive disc in Age Determination.
Background technology
The economic freshwater fish that Gui Shi China is important, because it is Fresh & Tender in Texture, nutritious, very popular without thorn between flesh again.Meanwhile, mandarin fish belongs to typical Piscivorous fishes in China's poisons in freshwater, is the top consumer in aquatic ecosystem, plays an important role adjustment and maintenance ecosystem balance.But in the last few years due to reasons such as overfishing, living environment deterioration and habitat habitat loss, mandarin fish natural resources were generally on a declining curve.The Age Determination of fish is bases of stock of fish Scientific evaluation, is also one of basic foundation of analysis and inspection dynamics of fish population trend, and the growth of fish individual, breeding, population supplement and the calculating of the death rate, all need to have accurate age data.Therefore, be a particularly important basic work to the precise Identification at mandarin fish age.
Fish Age Determination has longer research history, just starts as far back as 18th century scientist bony structures qualification fish age utilizing fish.The fish age is method the most traditional and conventional to utilize the wheel line in Yu Ti sclerous tissues to identify.At present, there are scale, otolith, vertebrae, nucleus, fin ray and cleithrum etc. in the hard tissue material being usually used in Age Determination, and wherein, the most frequently used is scale and otolith.Scale is considered to reliable Age Determination material for a long time, have draw materials simple, quantity is more, to advantages such as fish bulk damage are little, but because fish body has heavy absorbing phenomenon to scale, and scale is by body surface, wearing and tearing to a certain degree can be received in growth course, therefore for fish at advanced age, qualification result is often on the low side than actual age.Except annual ring mark, the scale of fish also there will be other and take turns line, as accessory whorl, children's wheel etc., interference can be caused to the qualification work at age.Otolith is present in Teleostei inner ear membranous labyrinth, plays the sclerous tissues of balance orientation and sense of hearing effect, form primarily of calcium carbonate.The general comparatively scale of the acquisition of otolith is complicated, but due to the continuation of otolith deposition, be present in the inner ear of fish in addition, be subject to the ectoskeletal protection of head, the uniformity of its growth and the complete performance of form are well protected.Therefore the annual ring of otolith and the sun can have recorded the fish course history of life exactly.At present, otolith is most widely used Age Determination material, is also the most accurate Age Determination material.
At present, the material mainly nucleus of mandarin fish Age Determination employing.Mandarin fish scale is tiny, close band and to dredge band not obvious, and often occur accessory whorl and reproduction wheel, winter wheel and false wheel are difficult to distinguish, and bring difficulty to Age Determination, therefore, scale is rarely used in the Age Determination of mandarin fish.Though annual ring is obvious on mandarin fish nucleus, there is dark-coloured herring bone in quite a few individuality in broadband, and the Besselian year layer that herring bone produces thus often can over-evaluate the real age of mandarin fish.
Mandarin fish sagitta is creamy white, translucent.Whole otolith is flat peltate, and interior lateral surface is uneven, and medial surface is outwardly, and lateral surface caves inward; Core is near lateral surface, be in the centre position of notopodium and siphonal lobe, comparatively obvious groove is together had between medial surface notopodium and siphonal lobe, above-mentioned contour structures causes extreme difficulties to Age Determination, the core of otolith and the wheel line of surrounding often can not intactly be presented on same abrasive disc, accurately cannot carry out Age Determination.Therefore, this method is by choosing mandarin fish sagitta as Age Determination material, abrasive disc is carried out to its cross section, ensure that otolith core and surrounding wheel line present on same abrasive disc, by wheel line distribution characteristics chequered with black and white on interpretation otolith abrasive disc, overcome the interference of Besselian year layer to annual ring, improve definition and the identification degree of annual ring, thus reach the object at precise Identification mandarin fish age.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of mandarin fish otolith cross section abrasive disc, the abrasive disc this method solving the uneven otolith of interior lateral surface makes difficult point, ensure that otolith core and wheel line intactly appear on abrasive disc, effectively improve definition and the identification degree of mandarin fish annual ring, thus reach the object at precise Identification mandarin fish age.
A further object of the invention there are provided the application of a kind of mandarin fish otolith cross section abrasive disc in Age Determination.
In order to achieve the above object, the present invention takes following technical measures:
A kind of preparation method of mandarin fish otolith cross section abrasive disc: the steps include:
1. otolith cleaning: take out after otolith, first use distilled water flushing, then put into percent by volume be 75% alcohol clean up, be placed on after drying in dimethylbenzene and soak 24 hours, make it transparent.
2. determine center line: after otolith cleans up, under being placed on microscopic transmission light, find mandarin fish Nucleus in otolith, then draw through centronucleus the line that is parallel to otolith minor axis with pencil, as center line, when being convenient to grind, observe progress.
3. prune: carefully otolith two ends are pruned smooth with little scissors, make itself and centerline parallel, to ensure fastness when otolith is fixed.
4. roughly grind: clamp otolith with otolith fixture (patent No. ZL 201220386074.X), otolith two ends that 600# sand paper is polished, make flour milling and centerline parallel, two ends flour milling separately distance center line 1.0-1.5mm time, corase grind completes.5. fix first: get a fritter PUR (705A), be placed on the central authorities of slide one end, slowly heat with alcolhol burner and make it melt, clamp otolith with tweezers and it is inserted in PUR perpendicular to slide.PUR solidifies rear otolith and can be fixed on slide.
6. fine grinding: hold slide, be stained with otolith one faces down, and forefinger withstands the fixing position of otolith at slide another side, first uses 600
#silicon carbide paper polishing, sand paper water-wet when polishing, become grind limit observation.Probably only have during 0.5mm select 1000 gradually wait grinding distance center line
#, 2000
#with 3000
#sand papering, until polishing to using polishing paper polishing during 0.2-0.3mm, one end grinds complete.
7. unload: first end grind complete after, slide is cleaned out, slowly adds thermoset PUR with alcolhol burner, take out otolith with tweezers, wash do not grind one end attachment PUR.
8. fix for two times: again heat (180 ~ 200 DEG C) slide and PUR is melted, clamp with tweezers one end that otolith do not grind, one end PUR ground vertically is sticked together on slide.
9. two polishings: polish according to the method for step (6), limit polishing is just placed on basis of microscopic observation, and after otolith core and wheel line are clear, use polishing paper polishing, buffed surface cleans out rear neutral gum mounting, obtains mandarin fish otolith cross section abrasive disc.
10. observe qualification: put under the microscope by the otolith abrasive disc made, observe and take turns line feature by the omnipotent video imaging apparatus (LY-WN-HP SUPER CCD) be connected with microscope and identify the age under incident light irradiates.
The application of mandarin fish otolith cross section abrasive disc in Age Determination, the steps include:
The otolith abrasive disc made is put under the microscope, observed by the CCD be connected with microscope under incident light irradiates and take turns line feature, record wheel line distance, take pictures and the interpretation age.According to the actual size of otolith cross section, otolith abrasive disc is amplified 10-40 doubly.Under incident light irradiates, otolith core is black, is in the center of otolith abrasive disc side (lateral surface), forms linear nucleus district.From otolith core, form the bright band zone staggered with blanking bar towards the medial surface of otolith, notopodium and siphonal lobe side, bright band is wider, the brown color be translucent, formation time be spring to autumn, being referred to as the summer takes turns 8; Blanking bar is narrower, in opaque black, is formed at winter, is referred to as winter wheel 7.Summer wheel 8 and winter wheel 7 alternately occur, forms a complete annual ring, count winter wheel 7, on abrasive disc, winter wheel 7 counts and is age size.
Compared with prior art, the present invention has the following advantages:
Mandarin fish otolith profile is obviously different from other most of fish, and its main feature is that interior lateral surface is uneven, and medial surface is outwardly, and lateral surface caves inward; Core, near lateral surface, is in the centre position of notopodium and siphonal lobe, has together comparatively significantly groove between medial surface notopodium and siphonal lobe.Above-mentioned contour structures causes extreme difficulties to Age Determination, and the core of otolith and the wheel line of surrounding often can not intactly be presented on same abrasive disc, therefore accurately cannot carry out Age Determination.The abrasive disc that the invention solves the uneven otolith of interior lateral surface makes difficult point, ensures that otolith core and wheel line intactly appear on abrasive disc, provides a kind of new method for otolith presents interior lateral surface rough fish Age Determination.The method has the advantage of more clear, the annual ring of wheel line more readability compared with conventional method; Secondly, otolith abrasive disc makes simple, and can complete an abrasive disc for every 4-6 minute, and otolith core and surrounding wheel line present on same abrasive disc to take the production method of cross section abrasive disc to ensure, are convenient to interpretation annual ring; Again, on cross section abrasive disc, the side of otolith core deflection abrasive disc, presents from otolith core the wheel line that light and shade replaces, and summer wheel and winter take turns high-visible, can overcome the interference of Besselian year layer to annual ring, ensure that the accuracy that annual ring is identified.
Accompanying drawing explanation
Fig. 1 is a kind of medial and lateral facial contour schematic diagram of mandarin fish sagitta.
Wherein, 1 is otolith notopodium, and 2 is otolith siphonal lobe, and 3 is the groove on otolith medial surface, and 4 is otolith core, and 5 is otolith center line, and 6 is the thickness of otolith abrasive disc.
Fig. 2 is total mandarin fish sagitta cross section abrasive disc after being.
Wherein, 7 is winter wheel (arrowband), represents annual ring, and 8 is summer wheel (broadband).
Detailed description of the invention
Be described in detail to of the present invention below in conjunction with concrete diagram:
Embodiment 1: a kind of preparation method of mandarin fish otolith cross section abrasive disc: the steps include:
1. otolith cleaning: the brainpan opening mandarin fish, from sacculus, sagitta is taken out with tip tweezers, be placed on and fill in the culture dish of clear water, the mucous membrane on otolith top layer is peeled gently off with dissecting needle, being placed on percent by volume after cleaning up is clean to remove the grease on otolith top layer in the alcohol of 75%, be placed on after drying in dimethylbenzene and soak 24 hours, make it transparent.
2. determine center line: after 24 hours, the otolith soaked is taken out from xylene solution, dry for subsequent use after rinsing out with clear water the dimethylbenzene that otolith shows to remain.Then be placed on microscope carrier by otolith, lateral surface, finds mandarin fish Nucleus in otolith 4 upward in transmitted light, then draws through centronucleus the center line 5 that is parallel to otolith minor axis with pencil, to observe progress when grinding.
3. prune: carefully otolith two ends are pruned smooth with little scissors, make it parallel with center line 5, to ensure fastness when otolith is fixed.
4. roughly grind: clamp otolith, 600 with otolith fixture
#sand paper to be polished otolith two ends, makes flour milling and centerline parallel, two ends flour milling separately distance center line 1.0-1.5mm time, corase grind completes.
5. fix first: get a fritter PUR, be placed on the central authorities of slide one end, slowly heat with alcolhol burner and make it melt, clamp the otolith pruned with tweezers, it is inserted in PUR perpendicular to slide.PUR solidifies rear otolith and can be fixed on slide.
6. polish: hold slide, be stained with otolith one faces down, forefinger withstands the fixing position of otolith at slide another side, first uses 600
#silicon carbide paper polishing, sand paper water-wet when polishing, become grind limit observation.Probably only have during 0.5mm select 1000 successively wait grinding distance center line
#, 2000
#with 3000
#sand papering, until polishing to 6 using polishing paper polishing during 0.2-0.3mm, one end grinds complete.
7. unload: first end grind complete after, slide is cleaned out, slowly adds thermoset PUR with alcolhol burner, take out otolith with tweezers, cleaning do not grind one end attachment PUR.
8. fix for two times: again heat slide and PUR is melted, clamp with tweezers one end that otolith do not grind, one end PUR ground vertically is sticked together on slide.
9. two polishings: polish according to the method for step (6), limit polishing is just placed on basis of microscopic observation, after otolith core and wheel line are clear, uses polishing paper polishing, buffed surface cleans out rear neutral gum mounting, obtains a kind of mandarin fish otolith cross section abrasive disc.
Embodiment 2: the application of a kind of mandarin fish otolith cross section abrasive disc in Age Determination, the steps include:
The otolith abrasive disc made is put under the microscope, observed by the CCD be connected with microscope under incident light irradiates and take turns line feature, record wheel line distance, take pictures and the interpretation age.According to the actual size of otolith cross section, otolith abrasive disc is amplified 10-40 doubly.Under incident light irradiates, otolith core is black, is in the center of otolith abrasive disc side (lateral surface), forms linear nucleus district.From otolith core, form the bright band zone staggered with blanking bar towards the medial surface of otolith, notopodium and siphonal lobe side, bright band is wider, the brown color be translucent, formation time be spring to autumn, being referred to as the summer takes turns 8; Blanking bar is narrower, in opaque black, is formed at winter, is referred to as winter wheel 7.Summer wheel 8 and winter wheel 7 alternately occur, forms a complete annual ring, count winter wheel 7, on abrasive disc, winter wheel 7 counts and is age size.
Otolith cross section abrasive disc wheel line is very clear, be easy to the interpretation age, and sentence read result accurately and reliably.Carry out Age Determination by said method to marking the 56 tail mandarin fishes (knowing age size accurately) of releasing, identical rate reaches 100%, further demonstrate that the accuracy at the inventive method qualification mandarin fish age.
Claims (2)
1. a preparation method for mandarin fish otolith cross section abrasive disc, the steps include:
1). otolith cleans: take out after otolith, first use distilled water flushing, then put into percent by volume be 75% alcohol clean up, be placed on after drying in dimethylbenzene and soak 24 hours;
2). determine center line: after otolith cleans up, under being placed on microscopic transmission light, find mandarin fish Nucleus in otolith, then draw the line that is parallel to otolith minor axis, as center line with pencil through centronucleus;
3). prune: otolith two ends are pruned smooth, makes itself and centerline parallel;
4). corase grind: clamp otolith, otolith two ends that sand paper is polished, make flour milling and centerline parallel, two ends flour milling separately distance center line 1.0-1.5mm time, corase grind completes;
5). fix first: by a little for hot melt rubber stick cutting, be placed on the central authorities of slide one end, make it melt with alcolhol burner heating, clamp otolith with tweezers and it inserted in PUR perpendicular to slide, PUR solidifies rear otolith and is fixed on slide;
6). fine grinding: hold slide, be stained with otolith one faces down, and forefinger withstands the fixing position of otolith at slide another side, first uses 600
#silicon carbide paper polishing, sand paper water-wet when polishing, grinds while observe; Probably only have during 0.5mm select 1000 gradually wait grinding distance center line
#, 2000
#with 3000
#sand papering, until polishing to using polishing paper polishing during 0.2-0.3mm, one end grinds complete;
7). unloading: first end grind complete after, slide is cleaned out, adds thermoset PUR with alcolhol burner, take out otolith with tweezers, wash do not grind one end attachment PUR;
8). secondary is fixed: again heat slide and PUR is melted, clamp one end that otolith do not grind, vertically sticked together on slide by one end PUR ground with tweezers;
9). secondary polish: according to step 6) method polishing, use polishing paper polishing, buffed surface cleans out rear neutral gum mounting.
2. the application of the otolith obtained by preparation method in mandarin fish Age Determination of a kind of mandarin fish otolith cross section abrasive disc according to claim 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103518655A (en) * | 2013-10-14 | 2014-01-22 | 中国水产科学研究院长江水产研究所 | Method for manufacturing section of xenophysogobio boulengeri otolith |
| CN103843708A (en) * | 2014-03-17 | 2014-06-11 | 上海海洋大学 | Method for constructing saury otolith physiological state analyzing model |
| CN104374615A (en) * | 2014-09-16 | 2015-02-25 | 中国水产科学研究院黑龙江水产研究所 | Schizothoracinae otolith slice production method |
| CN105028281B (en) * | 2015-08-17 | 2018-03-09 | 江苏省海洋水产研究所 | A kind of method that spindle and flat-sided shape fish quickly win sagitta |
| CN106813968B (en) * | 2017-02-09 | 2018-10-12 | 中国水产科学研究院黄海水产研究所 | A kind of preparation method of hairtail sagitta cross section |
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2013
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Patent Citations (8)
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| CA2088903A1 (en) * | 1993-02-04 | 1994-08-05 | Robert Brown | Method for identifying marked fish |
| CN1361475A (en) * | 2000-12-29 | 2002-07-31 | 中国科学院水生生物研究所 | Schlieren image processing process |
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