AU2021101269A4 - Method for determining age of sepia with sepia bone - Google Patents
Method for determining age of sepia with sepia bone Download PDFInfo
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- AU2021101269A4 AU2021101269A4 AU2021101269A AU2021101269A AU2021101269A4 AU 2021101269 A4 AU2021101269 A4 AU 2021101269A4 AU 2021101269 A AU2021101269 A AU 2021101269A AU 2021101269 A AU2021101269 A AU 2021101269A AU 2021101269 A4 AU2021101269 A4 AU 2021101269A4
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- 241000238370 Sepia Species 0.000 title claims abstract description 105
- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 80
- 238000000034 method Methods 0.000 title claims abstract description 28
- 230000007773 growth pattern Effects 0.000 claims abstract description 34
- 238000001035 drying Methods 0.000 claims abstract description 7
- 238000005498 polishing Methods 0.000 claims abstract description 7
- 238000002791 soaking Methods 0.000 claims description 9
- 241001441745 Sepia esculenta Species 0.000 claims description 8
- 239000012535 impurity Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 239000004927 clay Substances 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 3
- 210000003097 mucus Anatomy 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 2
- 241000238366 Cephalopoda Species 0.000 description 8
- 241000894007 species Species 0.000 description 7
- 230000012010 growth Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 2
- HBMJWWWQQXIZIP-UHFFFAOYSA-N silicon carbide Chemical compound [Si+]#[C-] HBMJWWWQQXIZIP-UHFFFAOYSA-N 0.000 description 2
- 229910010271 silicon carbide Inorganic materials 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 241000237852 Mollusca Species 0.000 description 1
- 241000472272 Sepiida Species 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 210000003323 beak Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 230000002842 otolith Effects 0.000 description 1
- 210000001265 otolithic membrane Anatomy 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/4833—Physical analysis of biological material of solid biological material, e.g. tissue samples, cell cultures
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/32—Polishing; Etching
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/366—Moulds; Demoulding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
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- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Optics & Photonics (AREA)
- Hematology (AREA)
- Biophysics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Farming Of Fish And Shellfish (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The present disclosure relates to the field of aquatic sciences, and in particular to
a method for determining an age of Sepia with a Sepia bone which includes grinding
of a longitudinal section of an internal shell, and observing and counting growth
patterns. The method includes the following main steps: drying, marking with lines,
cutting, grinding, polishing, marking with points, fixing, and counting growth patterns.
In order to solve the problems that an internal shell is easy to be broken during
grinding and growth patterns are difficult to observe and count, the present disclosure
provides a method that can quickly and accurately grind a section to the optimal plane
while accurately determining the number of growth patterns.
Description
[0001] The present disclosure relates to the field of aquatic sciences, and to a method for observing growth patterns on Sepia bones, and specifically, a method for determining an age of Sepia with a Sepia bone.
[0002] Sepia, an offshore species, is mainly distributed in the offshore areas of various countries, including China. This species has an annual output accounting for about 50% that of all offshore cephalopods, which has become one of the important fishing species in China's inshore fishery. As an important economic cephalopod at present, Sepia has attracted more attention from researchers regarding to basic biological and ecological research on the age, growth, migration, food intake, and the like thereof.
[0003] Age and growth are not only important research contents in cephalopod fishery biology and ecology, but also a basis for resource conservation and management of cephalopods. Cephalopods have the characteristics of short life span, fast growth, long spawning phase, complex population structure, etc., which makes an age estimation result of cephalopods not accurate enough by a traditional body-length frequency method. One of the most effective methods is to use growth patterns on hard tissues to estimate ages of cephalopods.
[0004] Otoliths, beaks, etc. are considered to be reliable materials for determining an age of Sepia. In past studies, researchers had tried to use a growth pattern on a cuticle on the surface of a Sepia bone to determine an age of Sepia. However, as an early growth pattern is difficult to interpret, the cuticle cannot be used for age determination.
[0005] The present disclosure is intended to provide a method for determining an age of Sepia with a Sepia bone, mainly including the following steps: drying, marking with lines, cutting, grinding, polishing, marking with points, fixing, and counting growth patterns. A longitudinal section of the Sepia bone is ground and polished. A prepared Sepia bone is photographed with a stereoscopic microscope to ensure that the growth patterns can be read at any time in the future. The growth patterns are counted from bottom to top of the Sepia bone.
[0006] In order to achieve the above objective, the present disclosure provides a method for determining an age of Sepia with a Sepia bone, including the following steps:
[0007] (1) drying: taking out a Sepia bone and using a hot air gun to blow dry a surface of the Sepia bone;
[0008] (2) marking with lines: marking a center line on the Sepia bone and a parallel line 1 cm on the right of the center line;
[0009] (3) cutting: cutting the Sepia bone longitudinally along the marked line on the right side;
[0010] (4) grinding: grinding an obtained longitudinal section of the Sepia bone with a grinder, where, coarse grinding is conducted first and fine grinding is then conducted until the section is in alignment with the center line;
[0011] (5) polishing: polishing a ground longitudinal section of the Sepia bone to remove oil and impurities;
[0012] (6) marking with points: marking the surface of the Sepia bone from bottom to top;
[0013] (7) fixing: putting the polished Sepia bone into a rectangular mold with white super light clay for fixing; and
[0014] (8) observation of growth patterns: counting growth patterns from bottom to top of the Sepia bone until all growth patterns are counted.
[0015] Further, before the drying in step (1), the Sepia bone may be pre-treated by the following steps:
[0016] A. collection: using a scalpel and scissors to carefully taking out an internal shell in Sepia, where, a Sepia bone is prevented from being broken and fractured;
[0017] B. soaking: soaking an obtained Sepia bone in alcohol to remove mucus and impurities on a surface of the Sepia bone;
[0018] C. cleaning: using a brush and water to clean the surface of the Sepia bone; and
[0019] D. storage: storing a cleaned Sepia bone in a refrigerator at -20°C.
[0020] Further, the counting growth patterns in step (8) includes:
[0021] I. photographing: photographing the growth patterns with a CCD under 1x objective lens of a stereoscopic microscope, where, marks spaced by 2 cm on the
Sepia bone are photographed;
[0022] II. image stitching: using image processing software to stitch all images acquired for the Sepia bone together, where, two adjacent images are stitched together according to repeated marks on the two images to ensure accuracy of stitching; and
[0023] III. counting: using image processing software on a computer to open a stitched image, and counting according to the image.
[0024] Further, in the soaking step, the soaking may be conducted for 60 min with a % alcohol solution.
[0025] Further, in the washing step, the washing may be conducted until a resulting liquid after washing is no longer turbid.
[0026] Further, the marking with points in step (6) may refer to giving a point mark every 2 cm.
[0027] Further, the Sepia maybe Sepia esculenta from West Africa.
[0028] Definitions:
[0029] "Sepia" is a general term of mollusca, cephalopods, and other species of the order Sepiida, which is one of the important fishing species in China and an important marine economic species, and plays an important role in the ecosystem. It should be understood that the present disclosure is not limited to specific species of Sepia, for example, Sepia esculenta from West Africa.
[0030] "Growth pattern" refers to fine wavy horizontal patterns, which can increase with the growth.
[0031] "Image processing software" is a general term for various application software used to process image information. Professional image processing software includes Adobe's photoshop. Those skilled in the art can select appropriate image processing software according to actual conditions, and the appropriate image processing software is also included in the protection scope of the present disclosure. The image processing software of the present disclosure is not limited to photoshop.
[0032] The present disclosure has the following advantages:
[0033] 1. The method for determining an age of Sepia with a Sepia bone provided in the present disclosure can achieve easy collection of an internal shell, convenient grinding, and stable information storage. Moreover, the method for determining an age of Sepia with a Sepia bone is direct, reliable, and accurate.
[0034] 2. The present disclosure selects a longitudinal section of a Sepia bone, which improves the readability of growth patterns and the accuracy of counting.
[0035] 3. In the method of the present disclosure, a photographing system is adopted for counting growth patterns, which greatly improves the convenience of counting growth patterns; and a prepared section is photographed with a stereoscopic microscope to ensure that the growth patterns can be read at any time in the future.
[0036] 4. The present disclosure solves the problem that growth patterns on a Sepia bone are difficult to observe and count (a large number of growth patterns at an initial stage of growth are under a cuticle), and provides a method that can quickly and accurately grind a section to the optimal plane while ensuring a high success rate of Sepia bone grinding, and can also accurately determine the number of growth patterns.
[0037] With reference to the accompanying drawings and specific examples of the present disclosure, those skilled in the art will better understand the above and other objectives, advantages, and features of the present disclosure.
[0038] FIG. 1 is a schematic diagram illustrating the morphology of a bone of Sepia esculenta from West Africa according to the present disclosure.
[0039] FIG. 2 is a schematic stitched image illustrating growth patterns on the bone of Sepia esculenta from West Africa according to the present disclosure.
[0040] The raw materials and devices used in the examples are well known to those skilled in the art, which are all commercially available or easily obtained or prepared. The specific implementations provided in the present disclosure will be further described in detail below with reference to examples.
[0041] Example: Method for determining an age of Sepia esculenta from West Africa with a bone thereof
[0042] As shown in FIG. 1 and FIG. 2, FIG. 1 is a schematic diagram illustrating the morphology of a bone of Sepia esculenta from West Africa,
[0043] FIG. 2 is a schematic stitched image illustrating growth patterns on the bone of Sepia esculenta from West Africa.
[0044] Operation steps in this example were as follows:
[0045] (1) collection: a scalpel and scissors were used to carefully take out an internal shell in Sepia, where, a Sepia bone was prevented from being broken and fractured;
[0046] (2) soaking: an obtained Sepia bone was soaked in alcohol to remove mucus and impurities on a surface of the Sepia bone;
[0047] (3) cleaning: a brush and water were used to clean the surface of the Sepia bone;
[0048] (4) storage: a cleaned Sepia bone was stored in a refrigerator at -20°C;
[0049] (5) drying: a Sepia bone was taken out and a surface of the Sepia bone was blow dried with a hot air gun;
[0050] (6) marking with lines: the Sepia bone was marked with a center line and a parallel line 1 cm on the right of the center line;
[0051] (7) cutting: the Sepia bone was cut longitudinally along the marked line on the right side;
[0052] (8) grinding: an obtained longitudinal section of the Sepia bone was ground with a grinder, where, during a grinding process, the section was ground successively with 240, 600, 1,200, 2,500 silicon carbide paper until the section was in alignment with the center line;
[0053] (9) polishing: a ground surface was polished with a flannel added with a 0.05 m alumina agent and water to remove surface scratches and excess impurities caused by the silicon carbide paper, thus completing the grinding on the longitudinal section of the Sepia bone;
[0054] (10) marking with points: marking the surface of the Sepia bone from bottom to top at an interval of 2 cm;
[0055] (11) fixing: the polished Sepia bone was put into a rectangular mold with white super light clay for fixing;
[0056] (12) photographing: the growth patterns were photographed with a CCD under 1x objective lens of a stereoscopic microscope, where, marks spaced by 2 cm on the Sepia bone were photographed;
[0057] (13) image stitching: image processing software was used to stitch all images acquired for the Sepia bone together, where, two adjacent images were stitched together according to repeated marks on the two images to ensure accuracy of stitching; and
[0058] (14) counting: image processing software on a computer was used to open a stitched image, and growth patterns were counted according to the image.
[0059] Through the above steps, the age information of a Sepia individual can be effectively studied and read. In the method of the present disclosure, a Sepia bone that is stable in shape, easy to collect, and easy to grind is used to determine an age of Sepia. The method includes the following steps: preparing a Sepia bone, and observing and counting growth patterns on an internal shell. Reading the number of growth patterns on a longitudinal section of a Sepia bone to determine an age of Sepia is a new method for directly determining an age of Sepia.
[0060] The preferred examples of the present disclosure have been described in detail above, but the present disclosure is not limited to these examples. Those skilled in the art can make various equivalent variations or substitutions without departing from the spirit of the present disclosure, and these equivalent variations or substitutions are all included in the scope defined by the claims of this application.
Claims (7)
- CLAIMS: 1. A method for determining an age of Sepia with a Sepia bone, comprising the following steps: (1) drying: taking out a Sepia bone and using a hot air gun to blow dry a surface of the Sepia bone; (2) marking with lines: marking a center line on the Sepia bone and a parallel line 1 cm on the right of the center line; (3) cutting: cutting the Sepia bone longitudinally along the marked line on the right side; (4) grinding: grinding an obtained longitudinal section of the Sepia bone with a grinder, wherein, coarse grinding is conducted first and fine grinding is then conducted until the section is in alignment with the center line; (5) polishing: polishing a ground longitudinal section of the Sepia bone to remove oil and impurities; (6) marking with points: marking the surface of the Sepia bone from bottom to top; (7) fixing: putting the polished Sepia bone into a rectangular mold with white super light clay for fixing; and (8) observation: counting growth patterns from bottom to top of the Sepia bone until all growth patterns are counted.
- 2. The method for determining an age of Sepia with a Sepia bone according to claim 1, wherein, before the drying in step (1), the Sepia bone is pre-treated by the following steps: A. collection: using a scalpel and scissors to carefully taking out an internal shell in Sepia, wherein, a Sepia bone is prevented from being broken and fractured; B. soaking: soaking an obtained Sepia bone in alcohol to remove mucus and impurities on a surface of the Sepia bone; C. cleaning: using a brush and water to clean the surface of the Sepia bone; and D. storage: storing a cleaned Sepia bone in a refrigerator at -20°C.
- 3. The method for determining an age of Sepia with a Sepia bone according to claim 1, wherein, the counting growth patterns in step (8) comprises: I. photographing: photographing the growth patterns with a CCD under 1x objective lens of a stereoscopic microscope, wherein, marks spaced by 2 cm on the Sepia bone are photographed; II. image stitching: using image processing software to stitch all images acquired for the Sepia bone together, wherein, two adjacent images are stitched together according to repeated marks on the two images to ensure accuracy of stitching; and III. counting: using image processing software on a computer to open a stitched image, and counting according to the image.
- 4. The method for determining an age of Sepia with a Sepia bone according to claim 2, wherein, in the soaking step, the soaking is conducted for 60 min with a 75% alcohol solution.
- 5. The method for determining an age of Sepia with a Sepia bone according to claim 2, wherein, in the washing step, the washing is conducted until a resulting liquid after washing is no longer turbid.
- 6. The method for determining an age of Sepia with a Sepia bone according to claim 1, wherein, the marking with points in step (6) refers to giving a point mark every 2 cm.
- 7. The method for determining an age of Sepia with a Sepia bone according to claim 1, wherein, the Sepia is Sepia esculenta from West Africa.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010506013.1A CN111693527A (en) | 2020-06-05 | 2020-06-05 | Method for identifying day age of cuttlefish by using cuttlefish bone |
| CN202010506013.1 | 2020-06-05 |
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| Publication Number | Publication Date |
|---|---|
| AU2021101269A4 true AU2021101269A4 (en) | 2021-05-06 |
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| AU2021101269A Ceased AU2021101269A4 (en) | 2020-06-05 | 2021-03-11 | Method for determining age of sepia with sepia bone |
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| Country | Link |
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| CN (1) | CN111693527A (en) |
| AU (1) | AU2021101269A4 (en) |
| LU (1) | LU102652B1 (en) |
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| CN112825797B (en) * | 2020-12-31 | 2022-12-30 | 上海海洋大学 | Construction method of iris cuttlefish horny jaw growth model based on marine environmental factors |
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| CN105738363B (en) * | 2014-12-12 | 2018-05-25 | 上海海洋大学 | A kind of method using Angle Overlap Model identification siphonopods age in days |
| CN104931665A (en) * | 2015-05-26 | 2015-09-23 | 上海海洋大学 | Construction method of vertebral growth characteristic analytical model for mackerel pike |
| CN106970079B (en) * | 2017-02-27 | 2019-12-27 | 上海海洋大学 | Method for identifying day age of teleost by using inner shell |
| CN107279017B (en) * | 2017-07-12 | 2020-04-07 | 上海海洋大学 | Method for researching growth of squid individual by using inner shell forenail |
| CN107328778A (en) * | 2017-07-12 | 2017-11-07 | 上海海洋大学 | A kind of method that utilization gastric mill identifies the Shrimp waste age |
| CN109588348A (en) * | 2018-12-19 | 2019-04-09 | 上海海洋大学 | A method of measuring and calculating South China Sea Symeplectoteuthis oualaniensis individual growth and maturity based on Angle Overlap Model form |
| CN109864020B (en) * | 2019-01-16 | 2021-05-04 | 上海海洋大学 | Method for measuring and calculating age and individual growth size of south sea iris cuttlefish based on external form of otolith |
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| Publication number | Publication date |
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| CN111693527A (en) | 2020-09-22 |
| LU102652B1 (en) | 2021-09-13 |
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