CN111838035A - Method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure - Google Patents
Method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure Download PDFInfo
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- CN111838035A CN111838035A CN201910938502.1A CN201910938502A CN111838035A CN 111838035 A CN111838035 A CN 111838035A CN 201910938502 A CN201910938502 A CN 201910938502A CN 111838035 A CN111838035 A CN 111838035A
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- 241000392375 Sinonovacula constricta Species 0.000 title claims abstract description 92
- 230000003204 osmotic effect Effects 0.000 title claims abstract description 44
- 238000012216 screening Methods 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 30
- 230000015784 hyperosmotic salinity response Effects 0.000 title claims abstract description 22
- 238000012258 culturing Methods 0.000 claims abstract description 6
- 210000000087 hemolymph Anatomy 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 238000002474 experimental method Methods 0.000 claims description 9
- 241000227752 Chaetoceros Species 0.000 claims description 7
- 241000206751 Chrysophyceae Species 0.000 claims description 7
- 238000005259 measurement Methods 0.000 claims description 7
- 210000003205 muscle Anatomy 0.000 claims description 5
- 238000012163 sequencing technique Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 4
- 238000007710 freezing Methods 0.000 claims description 4
- 241000195493 Cryptophyta Species 0.000 claims description 3
- 238000009360 aquaculture Methods 0.000 claims 1
- 244000144974 aquaculture Species 0.000 claims 1
- 238000009530 blood pressure measurement Methods 0.000 abstract 1
- 238000009395 breeding Methods 0.000 description 8
- 230000001488 breeding effect Effects 0.000 description 8
- 235000020639 clam Nutrition 0.000 description 4
- 235000015170 shellfish Nutrition 0.000 description 4
- 241000237852 Mollusca Species 0.000 description 2
- 241000237538 Solenidae Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 241000317942 Venturia <ichneumonid wasp> Species 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 241000894007 species Species 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/50—Culture of aquatic animals of shellfish
- A01K61/54—Culture of aquatic animals of shellfish of bivalves, e.g. oysters or mussels
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/90—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination
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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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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Zoology (AREA)
- Marine Sciences & Fisheries (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for screening sinonovacula constricta with strong salt tolerance by utilizing osmotic pressure, which comprises the steps of culturing sinonovacula constricta in the same culture environment, grouping salinity stress treatment, osmotic pressure measurement and final screening treatment. The sinonovacula constricta screening method can effectively screen out sinonovacula constricta with good salt tolerance, is simple, does not need complex equipment, has high screening efficiency and has high commercial value.
Description
Technical Field
The invention belongs to the technical field of new sinonovacula constricta species cultivation, and particularly relates to a method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure.
Background
Sinonovacula constricta Lamarck, commonly known as razor clam, is a shellfish of the order Venturia Bishell class, lives in the ocean, and is a common seafood food material. The shell is crisp, thin and in a shape of a long flat square, and an oblique concave ditch is formed from the top of the shell to the abdominal edge, so that the razor clam is named. It likes living in the gulf mud tidal flat and is a common mollusk in China coastal. The Sinonovacula constricta Lamarck belongs to the phylum mollusca, class Lamellibranchia, class Heteroodonta, order Veneruleales, family Solonovacula, and is one of four cultured shellfish in China. The method is widely distributed in China and the coastal areas of Japan, is mainly distributed in the coastal beaches of south and north China, and has quite rich yield. The Ningbo coastal zone has the advantage of being unique in cultivating razor clams, is a Ningbo popular sea special product, faces the three gulfs, has a large amount of fresh water injected throughout the year, is proper in salt water, is rich in bait, is mainly coated with silt, so that the razor clams grow fast, are large in individuals, tender and fat in meat, white in color and fresh in taste, and are recorded according to the Qing Ninghai county record: razor clam, genus Lamiophlomis, called razor clam field, a species of the field, is narrow and long like the middle finger, a Xishi tongue, also called Mei.
In the aspect of the existing technology for breeding new sinonovacula constricta species, due to the problem of individual characteristics of sinonovacula constricta, part of sinonovacula constricta cannot adapt to the change of salinity of a water body, so that the problem of great difficulty in breeding and breeding sinonovacula constricta is solved, a mature system is not provided in the aspect of screening sinonovacula constricta breeding varieties at present, and breeding is mostly carried out through complicated manual screening, so that the method is low in efficiency and general in twenty-effect.
Disclosure of Invention
The invention aims to provide a method for screening sinonovacula constricta with strong salt tolerance by utilizing osmotic pressure, which solves the problems of low breeding efficiency and poor effect in breeding new sinonovacula constricta species at present.
In order to solve the problems, the invention provides a method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure, which mainly comprises the following steps:
(1) pretreatment:
taking healthy Sinonovacula constricta adult shells without damage on the surfaces, temporarily culturing in a circulating culture aquarium for one week, continuously inflating to adapt to the environment, and feeding mixed algae of chrysophyceae and chaetoceros 2 times every morning and evening.
(2) Grouping salinity stress treatment:
setting a plurality of experimental groups with different water body salinity, putting 50 sinonovacula constricta treated in the step (1) into each group, and performing a salinity stress experiment;
(3) Osmolarity measurement
After the experiment stress, detecting the sinonovacula constricta in each group at 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96h respectively, quickly extracting hemolymph from the adductor muscle of the sinonovacula constricta by using a 1ml sterile syringe, moving the hemolymph into a sample tube, measuring the hemolymph osmotic pressure by using an osmometer, and recording the time point when the hemolymph osmotic pressure in each sinonovacula constricta reaches balance;
(4) sinonovacula constricta Lamarck screening method with strong salt tolerance
And (4) sequencing the time points of the sinonovacula constricta osmotic pressure reaching the equilibrium point recorded in the step (3), selecting the sinonovacula constricta with the speed of 10 times before reaching in each group, and taking out the sinonovacula constricta, thereby finishing the screening.
Preferably, in the step (1), the salinity of the water body in the circulating culture aquarium is 20.47 +/-0.94, the pH is 8.1 +/-0.2, and the water temperature is 25.12 +/-3.86 DEG C
Preferably, in the step (1), the feeding amount of the chrysophyceae and the chaetoceros is 3-5% of the total mass of the sinonovacula constricta.
Preferably, in the step (2), the number of the experimental groups is 5, and the salinity of each experimental group is 5, 15, 20, 25 and 35.
Preferably, in the step (3), the osmometer is a Gootec 3000 freezing-point osmometer available from Gootec GmbH of Germany.
The technical scheme has the following beneficial technical effects:
1. the method is simple to operate, does not need complicated steps and complex equipment, has low requirements on operators, and is suitable for most kinds of sinonovacula constricta;
2. the sinonovacula constricta salt tolerance screened by the method is strong, is suitable for farmers needing new sinonovacula constricta species cultivation, and has strong adaptability;
3. compared with the traditional screening method, the method provided by the invention has higher speed, is also suitable for rapid breeding and cultivation, can more scientifically and rapidly screen the sinonovacula constricta with strong environmental adaptability, and has sufficient practical value and commercial value.
Drawings
FIG. 1 is a graph of osmotic pressure versus time for each group in example 1;
in the figure, the horizontal axis represents time, and the vertical axis represents osmotic pressure.
Detailed description of the preferred embodiments
The invention is further described with reference to specific examples.
The following examples are not provided to limit the scope of the present invention, nor are the steps described to limit the order of execution. Modifications of the invention which are obvious to those skilled in the art in view of the prior art are also within the scope of the invention as claimed.
Example one
The invention discloses a method for screening sinonovacula constricta with strong salt tolerance by utilizing osmotic pressure, which comprises the following steps
(1) Pretreatment:
taking healthy Sinonovacula constricta adult shells without damage on the appearance, temporarily culturing the Sinonovacula constricta adult shells in a circulating culture aquarium for one week respectively, continuously aerating the Sinonovacula constricta adult shells to adapt to the environment, and feeding the Sinonovacula constricta adult shells with the golden algae and the Chaetoceros for 2 times every morning and evening, wherein the input amount is 5 percent of the mass of the Sinonovacula constricta adult shells;
(2) grouping salinity stress treatment:
setting 5 experimental groups of 5, 15, 20, 25 and 35 water salinity, putting 50 sinonovacula constricta treated in the step (1) into each experimental group, and performing a salinity stress experiment;
(3) osmolarity measurement
After the experimental stress, the sinonovacula constricta in each group is detected at 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96h respectively, hemolymph is quickly extracted from the adductor muscle of the sinonovacula constricta by using a 1ml sterile syringe, the hemolymph is moved into a sample tube, the hemolymph osmotic pressure is measured by using a Gootec 3000 freezing point osmometer which is produced by the Germany Gootec GmbH company, and the time point when the hemolymph osmotic pressure in each sinonovacula constricta reaches balance is recorded;
(4) sinonovacula constricta Lamarck screening method with strong salt tolerance
And (4) sequencing the time points of the sinonovacula constricta osmotic pressure reaching the equilibrium point recorded in the step (3), selecting and removing the sinonovacula constricta with the speed of 10 before the sinonovacula constricta osmotic pressure reaches the equilibrium point in each group, and finishing the screening.
Example two:
the invention discloses a method for screening sinonovacula constricta with strong salt tolerance by utilizing osmotic pressure, which comprises the following steps
(1) Pretreatment:
taking healthy Sinonovacula constricta adult shells without damage on the appearance, temporarily culturing the Sinonovacula constricta adult shells in a circulating culture aquarium for one week respectively, continuously aerating the Sinonovacula constricta adult shells to adapt to the environment, and feeding the Sinonovacula constricta adult shells with the golden algae and the Chaetoceros for 2 times every morning and evening, wherein the input amount is 5 percent of the mass of the Sinonovacula constricta adult shells;
(2) grouping salinity stress treatment:
setting 5 experimental groups of 5, 15, 20, 25 and 35 water salinity, putting 50 sinonovacula constricta treated in the step (1) into each experimental group, and performing a salinity stress experiment;
(3) osmolarity measurement
After the experimental stress, the sinonovacula constricta in each group is detected at 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96h respectively, hemolymph is quickly extracted from the adductor muscle of the sinonovacula constricta by using a 1ml sterile syringe, the hemolymph is moved into a sample tube, the hemolymph osmotic pressure is measured by using a Gootec 3000 freezing point osmometer which is produced by the Germany Gootec GmbH company, and the time point when the hemolymph osmotic pressure in each sinonovacula constricta reaches balance is recorded;
(4) sinonovacula constricta Lamarck screening method with strong salt tolerance
And (4) sequencing the time points of the sinonovacula constricta osmotic pressure reaching the equilibrium point recorded in the step (3), selecting and removing the sinonovacula constricta with the speed of 10 before the sinonovacula constricta osmotic pressure reaches the equilibrium point in each group, and finishing the screening.
Example three:
the invention discloses a method for screening sinonovacula constricta with strong salt tolerance by utilizing osmotic pressure, which comprises the following steps
(1) Pretreatment:
taking healthy Sinonovacula constricta adult shellfish without damage on the surface, temporarily culturing in a circulating culture aquarium for one week, continuously aerating at the water temperature of 28.98 ℃ and the salinity of the water body of 21.41 and the pH of 8.3 to adapt to the environment, and feeding the mixed algae of chrysophyceae and chaetoceros 2 times every morning and evening, wherein the input amount is 5 percent of the mass of the Sinonovacula constricta adult shellfish;
(2) grouping salinity stress treatment:
setting 5 experimental groups of 5, 15, 20, 25 and 35 water salinity, putting 50 sinonovacula constricta treated in the step (1) into each experimental group, and performing a salinity stress experiment;
(3) osmolarity measurement
After the experimental stress, the sinonovacula constricta in each group is detected at 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96h respectively, hemolymph is quickly extracted from the adductor muscle of the sinonovacula constricta by using a 1ml sterile syringe, the hemolymph is moved into a sample tube, the hemolymph osmotic pressure is measured by using a Gootec 3000 freezing point osmometer which is produced by the Germany Gootec GmbH company, and the time point when the hemolymph osmotic pressure in each sinonovacula constricta reaches balance is recorded;
(4) sinonovacula constricta Lamarck screening method with strong salt tolerance
And (4) sequencing the time points of the sinonovacula constricta osmotic pressure reaching the equilibrium point recorded in the step (3), selecting and removing the sinonovacula constricta with the speed of 10 before the sinonovacula constricta osmotic pressure reaches the equilibrium point in each group, and finishing the screening.
The time between the internal osmotic pressure and the external osmotic pressure of the sinonovacula constricta in 5 experimental groups in example 1 was measured, and when the salinity was 35, the average time of osmotic pressure equilibrium was 6.5 hours, and when the salinity was 5, the average time of osmotic pressure equilibrium was 30 hours, and when the salinity was 15, 20, and 25, the average time of osmotic pressure equilibrium was 2 hours because the difference in osmotic pressure from the interior of the sinonovacula constricta was small.
After the experiment, the osmolarity of 10 sinonovacula constricta lamarck obtained by screening in example 1 was measured again under the condition that the experiment was stressed in the water body with salinity of 5, 15, 20, 25 and 35, and the osmolarity was measured in 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96 hours, respectively, and the average results of the measurements are shown in fig. 1.
As shown in figure 1, the sinonovacula constricta screened by the method only takes 4 hours in a high-salinity environment (with the salinity of 35) so as to facilitate the external osmotic pressure to reach balance, and in the measurement of example 1, the average time for the sinonovacula constricta to reach the osmotic pressure in the water body with the salinity of 35 is 6.5 hours; the method only takes 24 hours in the environment with low salinity (the salinity is 5), and the average time is greatly shortened compared with that in the example 1, so that the sinonovacula constricta salt tolerance screened by the method is better, and the method is simpler.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes or modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.
Claims (5)
1. A method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure is characterized by comprising the following steps:
(1) pretreatment:
taking healthy Sinonovacula constricta adult shells without damage on the surfaces, temporarily culturing in a circulating culture aquarium for one week, continuously inflating to adapt to the environment, and feeding mixed algae of chrysophyceae and chaetoceros 2 times every morning and evening;
(2) grouping salinity stress treatment:
setting a plurality of experimental groups with different water body salinity, putting 50 sinonovacula constricta treated in the step (1) into each group, and performing a salinity stress experiment;
(3) osmolarity measurement
After the experiment stress, detecting the sinonovacula constricta in each group at 0, 0.5, 1, 1.5, 2, 4, 8, 12, 24, 48, 72 and 96h respectively, quickly extracting hemolymph from the adductor muscle of the sinonovacula constricta by using a 1ml sterile syringe, moving the hemolymph into a sample tube, measuring the hemolymph osmotic pressure by using an osmometer, and recording the time point when the hemolymph osmotic pressure in each sinonovacula constricta reaches balance;
(4) Sinonovacula constricta Lamarck screening method with strong salt tolerance
And (4) sequencing the time points of the sinonovacula constricta osmotic pressure reaching the equilibrium point recorded in the step (3), selecting and removing the sinonovacula constricta with the speed of 10 before the sinonovacula constricta osmotic pressure reaches the equilibrium point in each group, and finishing the screening.
2. The method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure as claimed in claim 1, wherein in the step (1), the salinity of the water body in the circulating aquaculture aquarium is 20.47 ± 0.94, the pH is 8.1 ± 0.2, and the water temperature is 25.12 ± 3.86 ℃.
3. The method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure as claimed in claim 1, wherein in the step (1), the feeding amount of the chrysophyceae and the chaetoceros sp is 3% -5% of the total mass of the sinonovacula constricta.
4. The method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure as claimed in claim 1, wherein in the step (2), the number of experimental groups is 5, and the salinity of each experimental group is 5, 15, 20, 25 and 35 respectively.
5. The method for screening sinonovacula constricta with strong salt tolerance by using osmotic pressure as claimed in claim 1, wherein in the step (3), the osmometer is a Gootec 3000 freezing point osmometer available from Gootec GmbH of Germany.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113412804A (en) * | 2021-07-23 | 2021-09-21 | 上海海洋大学 | Method for judging survival of young eriocheir sinensis |
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| CN103548726A (en) * | 2013-10-23 | 2014-02-05 | 上海海洋大学 | Establishment and stock breeding method for sinonovacula constricta families |
| CN108323462A (en) * | 2018-02-08 | 2018-07-27 | 中国水产科学研究院黄海水产研究所 | A kind of breeding method of resistance to less salt chief blood clam seed |
| CN108812465A (en) * | 2018-07-02 | 2018-11-16 | 上海海洋大学 | One kind is hung razor clam seedling desalination method |
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- 2019-09-30 CN CN201910938502.1A patent/CN111838035A/en active Pending
Patent Citations (4)
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|---|---|---|---|---|
| CN101664007A (en) * | 2009-09-21 | 2010-03-10 | 宁波大学 | Sinonouacula hybridization breeding method |
| CN103548726A (en) * | 2013-10-23 | 2014-02-05 | 上海海洋大学 | Establishment and stock breeding method for sinonovacula constricta families |
| CN108323462A (en) * | 2018-02-08 | 2018-07-27 | 中国水产科学研究院黄海水产研究所 | A kind of breeding method of resistance to less salt chief blood clam seed |
| CN108812465A (en) * | 2018-07-02 | 2018-11-16 | 上海海洋大学 | One kind is hung razor clam seedling desalination method |
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| CN113412804A (en) * | 2021-07-23 | 2021-09-21 | 上海海洋大学 | Method for judging survival of young eriocheir sinensis |
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Application publication date: 20201030 |