CN114208736A - Method for improving artificial hatching rate of blood clam - Google Patents
Method for improving artificial hatching rate of blood clam Download PDFInfo
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- CN114208736A CN114208736A CN202111641175.7A CN202111641175A CN114208736A CN 114208736 A CN114208736 A CN 114208736A CN 202111641175 A CN202111641175 A CN 202111641175A CN 114208736 A CN114208736 A CN 114208736A
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- hatchability
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- 238000000034 method Methods 0.000 title claims abstract description 21
- 241001339782 Scapharca broughtonii Species 0.000 title claims description 30
- 230000012447 hatching Effects 0.000 title description 9
- 230000004720 fertilization Effects 0.000 claims abstract description 36
- 229910001424 calcium ion Inorganic materials 0.000 claims abstract description 19
- 241000224239 Tegillarca granosa Species 0.000 claims abstract description 18
- 239000013535 sea water Substances 0.000 claims abstract description 18
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910001425 magnesium ion Inorganic materials 0.000 claims abstract description 15
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 241000237506 Scapharca Species 0.000 claims abstract description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 12
- 210000004681 ovum Anatomy 0.000 claims description 11
- 102000002322 Egg Proteins Human genes 0.000 claims description 7
- 108010000912 Egg Proteins Proteins 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- 241000628923 Anadara sativa Species 0.000 claims description 4
- 230000001788 irregular Effects 0.000 claims description 4
- 235000015170 shellfish Nutrition 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 238000009287 sand filtration Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 13
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 abstract description 12
- 239000011575 calcium Substances 0.000 abstract description 12
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 2
- 230000012010 growth Effects 0.000 abstract description 2
- 235000013601 eggs Nutrition 0.000 description 13
- 239000000725 suspension Substances 0.000 description 7
- 238000009395 breeding Methods 0.000 description 5
- 230000001488 breeding effect Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- 230000004913 activation Effects 0.000 description 3
- 230000035558 fertility Effects 0.000 description 3
- 230000019100 sperm motility Effects 0.000 description 3
- 230000030120 acrosome reaction Effects 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000037427 ion transport Effects 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- CYYJCOXYBYJLIK-MCDZGGTQSA-L magnesium;[[[(2r,3s,4r,5r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-oxidophosphoryl] hydrogen phosphate Chemical compound [Mg+2].C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP([O-])(=O)OP(O)([O-])=O)[C@@H](O)[C@H]1O CYYJCOXYBYJLIK-MCDZGGTQSA-L 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000003359 percent control normalization Methods 0.000 description 1
- 210000004508 polar body Anatomy 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000008010 sperm capacitation Effects 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
Classifications
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention relates to the technical field of marine product production, and discloses a method for improving artificial hatchability of scapharca granosa, which comprises the steps of manually obtaining female and male gametes of scapharca granosa, carrying out artificial fertilization in a seawater environment added with calcium and magnesium ions, wherein the pH value of a water body is 8.0-8.2, the concentration of the added calcium ions is 200-200 mg/L, the concentration of the magnesium ions is 150-200mg/L, and the sperm-egg ratio is controlled to be 100:1-200: 1; the method has the advantages of simple operation, low cost, short operation time, and no influence on the development and growth of Arca granosa larvae.
Description
Technical Field
The invention relates to the field of marine product production, in particular to a method for improving artificial hatchability of blood clam.
Background
The Tegillarca granosa (Tegillarca granosa) is popular with consumers in China due to delicious meat quality and rich nutrition, and the artificial culture yield of the Tegillarca granosa is high, so that the Tegillarca granosa becomes an important mudflat culture economic shellfish in the southeast coastal area of China at present and is also an important component of a coastal mudflat marine ecosystem. However, as the tegillarca granosa adopts broadcast fertilization, the sperm and eggs are directly discharged into seawater in the breeding period and the whole processes of fertilization and growth and development are completed in an open water body, and the success rate of fertilization mainly depends on the interaction of gametes in the water body. In the 'influence of environmental factor change on sperm motility of blood clam', the Gejunquan et al disclose that sperm activation conditions affect fertility rate and hatchability, and preliminarily obtain the most suitable environmental conditions for blood clam sperm activation and movement as follows: the pH value is 8.5, the salinity is 25, and the water temperature is 30 ℃, and the water quality control method can be used as a selection and regulation reference for an optimal water environment factor for artificial induced spawning in industrial seedling production. Meanwhile, the ratio of sperms and eggs is also an important factor influencing the fertilization rate and the hatching rate; however, in natural environment, the fertilization rate and the hatching rate of the blood clam have been proved to be positively correlated with the ratio of the sperms and the eggs within certain limits.
At present, during industrial breeding of the tegillarca granosa, a higher sperm-egg ratio is often adopted to improve the fertility rate and the hatchability of the tegillarca granosa. However, the phenomenon of multifspermic fertilization can occur due to the overlarge proportion of sperms and ova, which can cause the fertilized ova to be incapable of normally developing, and on the other hand, the excessive sperms can also deteriorate the water quality, which causes the increase of the aberration rate, and the production of the offspring seeds is seriously restricted in the production practice.
Therefore, finding an efficient and convenient method for improving the artificial hatchability of the scapharca granosa is one of the key problems to be solved in the industrial breeding of the scapharca granosa.
Disclosure of Invention
Aiming at the defects that the conventional methods for breeding the blood clam of the blood clam easily cause the increase of the polyspermia fertilization rate of the blood clam and limit the hatchability of the blood clam, the invention provides a method for efficiently, conveniently and cheaply improving the artificial hatchability of the blood clam.
In order to solve the technical problem, the invention is solved by the following technical scheme:
a method for improving artificial hatchability of scapharca granosa comprises the following steps:
(1) shellfish cleaning: cleaning the surface dirt of the blood clam shells by sand filtration seawater;
(2) preparing a solution: respectively preparing a calcium chloride solution and a magnesium sulfate solution;
(3) obtaining an arca subcrenata gamete: acquiring gametes of the blood clam processed in the step (1), performing microscopic examination on the sperm-egg concentration, and discarding gametes with low sperm activity and irregular ovum shape;
(4) artificial fertilization: adding the calcium chloride solution and the magnesium sulfate solution prepared in the step (2) into filtered seawater; adding Arca granosa sperm and egg, and controlling sperm-egg ratio at 100:1-200:1
Preferably, the pH value of the seawater is 8.0-8.2.
Preferably, the concentration of the calcium chloride solution prepared in the step (2) is 10g/L, and the concentration of the magnesium sulfate solution is 10 g/L.
Preferably, the concentration of the calcium ions added in the step (4) is controlled to be 200-300mg/L, and the concentration of the magnesium ions is controlled to be 150-200 mg/L.
Preferably, the method for obtaining the tegillarca granosa gamete in the step (3) is drying in the shade, and specifically, the tegillarca granosa gamete treated in the step (1) is dried in the shade and exposed in the air at 23 ℃ for 12 hours, then, each tegillarca granosa individual is subjected to artificial induced spawning independently, and a male-female gamete suspension is collected; finally, the gamete quality is checked under a microscope, the sperm and egg concentration is measured, and the gamete suspension with low sperm motility and irregular ovum shape is discarded.
The acrosome reaction of the sperms depends on the internal flow of calcium ions outside the cells, and the capacitation of the sperms of the blood clams and the acrosome reaction can be promoted by increasing the concentration of the calcium ions in the water body; the increase of magnesium ions can increase the activity of calcium and magnesium ATP enzyme of sperms and increase the transport efficiency of sperms to calcium ions.
In addition, the increased efficiency of calcium ion transport within an egg cell may facilitate the release of cortical particles during fertilization, preventing multiple sperm from entering the egg. On the other hand, calcium and magnesium ions are used as messengers to participate in the formation of calcium waves in the egg cells in the fertilization process, and the calcium and magnesium ion concentration of the water body is increased to promote the activation of the egg cells.
Compared with the prior art, the invention has the beneficial effects that:
(1) the method is simple to operate, low in cost and more economical and practical;
(2) the fertilization rate of the blood clam is improved, simultaneously, the overhigh sperm-egg proportion is avoided, the multi-sperm fertilization rate and the abnormal rate are reduced, and the hatching rate is increased;
experiments prove that the artificial fertilization of the tegillarca granosa is carried out by adopting the method, the fertilization rate of the tegillarca granosa can be improved by 5-15% under the condition that the sperm-egg ratio is 100:1-200:1, the polyspermia fertilization rate can be reduced by 50-60%, and the overall hatchability is improved by 15-30%.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1: experiment for influence of calcium and magnesium ions in water body on fertilization of blood clam
Time: 7 months 2019, location: qingjiang base of research institute of marine aquaculture in Zhejiang province.
About 500 sexually mature blood clam shells (about 9g in weight) are taken, washed clean and then dried in the shade and exposed in the air at 23 ℃ for 12 hours. And then, independently placing each blood clam individual into a beaker filled with 500mL of filtered seawater for artificial spawning induction, and collecting the gamete suspension. Checking gamete quality under microscope, measuring sperm and egg concentration, and discarding gamete suspension with low sperm motility and irregular ovum shape. As shown in table 1, 15 experimental groups and 1 control group were set up in total to compare the effects of different calcium and magnesium ion concentrations on fertilization rate of blood clam, fertilization rate of sperm, and hatching rate (10 replicates per group).
TABLE 1 Experimental group settings
After filtering the collected gamete suspension of 300 blood clam shells (150 blood clam shells each), dividing each egg cell suspension into 16 parts, transferring 1 part into a common seawater bucket containing 10L, and transferring the other 15 parts into 15 experimental water buckets containing 10L (calcium and magnesium ions are added into seawater according to the table 1). Sperm and egg are added according to the sperm-egg ratio of 200:1 to carry out fertilization experiments. After gamete mixing for 1 hour, collect the ovum or zygote in each experimental group, count the zygote number (ovum is in the polar body discharge or cell division state) under the microscope, calculate the fertilization rate (zygote number/ovum cell number x 100%) and the polyspermia fertilization rate (polyspermia fertilization egg number/ovum cell number x 100%), count the hatchability rate (D-type larva number/fertilized ovum number x 100%) after 8 hours, and the experimental results are shown in table 2.
TABLE 2 fertilization rate, fertilization rate of polyspermy and hatching rate of two groups of Arca granosa
Note: the data are mean ± variance of the data sets.
This embodiment shows that, calcium and magnesium ions are added simultaneously, the concentration of calcium ions is controlled to be 200-.
Example 2: experiment for influence of calcium and magnesium ions in water body on fertilization of blood clam
Time: month 8 in 2021, site: qingjiang base of research institute of marine aquaculture in Zhejiang province.
About 500 Scapharca subcrenata (about 8g in weight) raised in acidified seawater (pH7.8) for 30 days was exposed to air drying at 23 ℃ for 12 hours after cleaning the surface dirt of the shell. And (3) independently placing the tegillarca granosa individuals into a beaker filled with 500mL of acidified seawater (pH7.8) for spawning induction, and collecting high-quality gamete suspension.
An experimental group (with calcium and magnesium ions added) and a control group were set to compare the difference in fertilization rates of arca subcrenata under two fertilization conditions (10 replicates per group).
Each collected gamete suspension (20 males and females) was divided into two parts, one part was transferred to a bucket containing 50L of acidified seawater (pH7.8), and the other part was transferred to a bucket containing 50L of experimental water (300 mg/L calcium ions and 150mg/L magnesium ions were added to seawater, pH 7.8). Fertilization experiments were performed with a sperm-egg ratio of 150: 1. The fertilization rate of the blood clam under the acidified seawater condition, the polyspermia fertilization rate and the hatching rate after 8 hours after 1 hour were counted under a microscope, and the results are shown in table 3.
TABLE 3 fertilization rate, fertilization rate of polyspermy and hatchability rate of two groups of Arca granosa under the acidified seawater condition
| Group of | Fertilization rate/%) | Rate of polyspermia/%) | Hatching rate/% |
| Control group | 63.7±8.3 | 18.3±4.3 | 34.6±2.1 |
| Treatment group | 74.7±5.6 | 10.6±3.1 | 43.5±1.8 |
Note: the data are mean ± variance of the data sets.
The fertility rate of the blood clam under the condition of the acidified seawater can be improved by 17.3%, the polyspermia fertilization rate can be reduced by 42.1%, and the hatchability rate can be remarkably increased by 25.7% by adding the calcium and magnesium ions, so that the method is proved to have an excellent effect of improving the hatchability rate of the artificial breeding of the blood clam.
The foregoing is a preferred embodiment of the present invention, and it will be apparent to those skilled in the art that various modifications and improvements can be made without departing from the spirit of the invention, and the scope of the invention shall be limited only by the appended claims.
Claims (6)
1. A method for improving artificial hatchability of blood clam is characterized by comprising the following steps:
(1) shellfish cleaning: cleaning the surface dirt of the blood clam shells by sand filtration seawater;
(2) preparing a solution: respectively preparing a calcium chloride solution and a magnesium sulfate solution;
(3) obtaining an arca subcrenata gamete: acquiring gametes of the blood clam processed in the step (1), performing microscopic examination on the sperm-egg concentration, and discarding gametes with low sperm activity and irregular ovum shape;
(4) artificial fertilization: adding the calcium chloride solution and the magnesium sulfate solution prepared in the step (2) into filtered seawater; adding Arca granosa sperm and egg, and controlling the ratio of sperm to egg to be 100:1-200: 1.
2. The method for improving artificial hatchability of scapharca granosa according to claim 1, wherein the seawater has a pH of 8.0-8.2.
3. The method for improving artificial hatchability of scapharca granosa according to claim 1 or 2, wherein the concentration of the calcium chloride solution prepared in the step (2) is 10g/L, and the concentration of the magnesium sulfate solution is 10 g/L.
4. The method for improving the artificial hatchability of the blood clam according to claim 3, wherein the method for obtaining the blood clam gametes in the step (3) is drying in the shade.
5. The method for improving artificial hatchability of scapharca granosa according to claim 3, wherein the concentration of the added calcium ions in the step (4) is controlled to be 200-300 mg/L.
6. The method for improving artificial hatchability of scapharca granosa according to claim 3, wherein the concentration of the magnesium ions added in the step (4) is controlled to be 150-200 mg/L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115088657A (en) * | 2022-07-29 | 2022-09-23 | 杭州师范大学 | Method for improving fertility rate and hatchability of old litopenaeus vannamei |
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