CN114258878A - In-situ spawning induction method for tridacna parents - Google Patents
In-situ spawning induction method for tridacna parents Download PDFInfo
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- 238000011065 in-situ storage Methods 0.000 title claims abstract description 63
- 241000512272 Tridacna Species 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 30
- 230000006698 induction Effects 0.000 title claims abstract description 28
- 235000020639 clam Nutrition 0.000 claims abstract description 75
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- XNOPRXBHLZRZKH-UHFFFAOYSA-N Oxytocin Natural products N1C(=O)C(N)CSSCC(C(=O)N2C(CCC2)C(=O)NC(CC(C)C)C(=O)NCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(CCC(N)=O)NC(=O)C(C(C)CC)NC(=O)C1CC1=CC=C(O)C=C1 XNOPRXBHLZRZKH-UHFFFAOYSA-N 0.000 description 2
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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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Abstract
The invention discloses an in-situ spawning method of tridacna parents. The method comprises the technical links of parent selection, in-situ induced spawning, larva inspection, induced spawning effect evaluation and the like. The method is different from a fully artificial breeding method of giant clams, the field semi-closed lagoons and other environments are screened as spawning inducing fields, in-situ spawning induction is carried out at night, the production process of artificial seedlings is simulated as much as possible, risks and enemies are avoided, and artificial spawning inducing type seedlings consistent with wild seedlings are obtained in batches; and (3) carrying out paternity recognition by combining a modern molecular marker technology, and evaluating the in-situ induced spawning effect. According to the giant clam larva breeding method, the floating characteristics of giant clams in the larva stage are fully utilized, large-area diffusion is realized by means of sea waves, ocean currents and the like, a part of giant clam larva gathering areas are formed, sufficient parents are provided for giant clam breeding in future, and technical support is provided for quick repair of giant clam resources in south China sea. The invention has the advantages of simple and easy operation, obvious effect and the like.
Description
Technical Field
The invention belongs to the technical field of shellfish breeding and breeding in marine agriculture, and particularly relates to an in-situ spawning method of tridacna parents.
Background
Giant clams are a general term for giant clams of the class of lamellibranchia of the phylum mollusca, and 12 species of giant clams are widely distributed in tropical coral reef sea areas from western pacific ocean to east coast of indian ocean africa. 8 types of Tridacna are distributed in China, and are Tridacna (Tridacan gigas), scale-free Tridacna (Tridacna derasa), scale Tridacna (Tridacna squamosa), long Tridacna (Tridacna maxima), nova Tridacna (Tridacna nova), safranina (Tridacna crocea) and trioyster (Hippopus) of Tridacna and tridactylus tridatus, respectively.
The lagoon, the fortunes and the reef plates of the reef island are original distribution sea areas of giant clams, the giant clams are very easy to catch due to the habit that the giant clams are fixedly living in the shallow transparent sea area of the tropical zone, the environment is deteriorated, people are caught, the number of giant clams is reduced sharply, the living giant clams are almost completely killed, scale-free giant clams are basically and functionally killed, and other giant clams are rare. In contrast, in 2016, the south sea ocean research institute of the academy of sciences breaks through the artificial breeding technology of tridacna, the healthy fries of 5 tridacna and 2 tridacna are cultivated, the proliferation and releasing of tridacna in the south sea of China are developed, and the increase of giant clam resource amount is realized. However, the quantity of artificial bottom sowing proliferation hardly meets the recovery demand of giant clam resources in south China, and therefore, development of an efficient spawning promotion technology is urgently needed, large-area in-situ proliferation of giant clam fries is realized by means of the forces of sea waves, ocean currents and the like, and the purpose of recovering giant clam resources is achieved.
Disclosure of Invention
The invention aims to provide an in-situ spawning method of tridacna parents, which is simple, convenient and easy to implement and has a remarkable effect.
The in-situ spawning induction method of tridacna parents is characterized by comprising the following steps:
a. gonad monitoring: finding a distribution group of wild or artificially transplanted giant clams by diving in a 5-30 m water-depth wave-free area environment of fields such as south-sea field reef sea areas, lagoons, ring reefs and the like in China, positioning, recording the site position of a parent group, and regularly observing the gonad development condition;
b. in-situ induced spawning: when the gonads mature, diving to a giant clam group distribution area, and injecting an oxytocic into a mantle for in-situ oxytocic;
c. larva examination: after the in-situ production promotion of the parents, collecting plankton in water at multiple points in the waters of lagoons, circular reefs and the like, and filtering large individual impurities; after fixing with alcohol, observing the number of larvae; then detecting the abundance and the number of the podosphaeria pan larvae; identifying whether giant clam larvae are giant clam offspring or not in-situ induced spawning offspring;
d. and (3) evaluating the spawning induction efficiency: after 6-9 months of induced spawning, checking whether giant clam seedlings of about 10-30mm are around lagoons and circular reefs where the in-situ induced spawning sites are located, taking back a plurality of individuals, carrying out paternity test, detecting the in-situ induced spawning efficiency, and calculating the number of offspring obtained by induced spawning.
Preferably, in the step a, the distribution group of tridacna can be a distribution group in a wild state or a distribution group of tridacna after artificial transplantation, but the tridacna must reach sexual maturity age to ensure that sperm eggs can be produced.
Preferably, in the step b, the oxytocin is a serotonin solution, and the dosage of the oxytocin is within the range of 0.3-0.5mg/kg of total weight of serotonin/tridacna.
Preferably, in the step c, the specific molecular marker refers to a composite PCR primer combination capable of distinguishing more than 3-5 giant clams simultaneously, and can effectively distinguish larva compositions and parent sources thereof. See, for example, Ma, H, Feng, H, Zhang, Y, Qin, Y, Xiao, Z, Li, J, Zhang, Y, Yu, Z, multiple specific PCR identification of a natural viral amplification in the South China Sea: a useful tool for application in viral storage management and for information identification.
Preferably, in the step d, the feasibility of the in-situ spawning induction efficiency extremely for resource recovery is evaluated through the number and variety of tridacna per unit area and a parent-child recognition technology.
Preferably, the step a is positioning by using GPS.
Preferably, the regular observation of the gonadal development is realized by a method combining visual observation and microsampling.
Preferably, in the step b, during in-situ catalysis, only one tridacna is selected for injection each time in order to obtain pure tridacna resources; in order to guarantee the mating efficiency of gametes, the spawning induction time is controlled within 1 h; in order to reduce the eating probability of wild enemy gamete, the spawning induction time is selected to be 8-10 points at night; in order to improve the use efficiency of the eggs, the discharged eggs are collected firstly and then scattered to be combined with newly discharged sperms, so that fertilized eggs are promoted to drift along with sea waves.
Preferably, the larva inspection of step c is: after the parent is induced to spawn in situ for 24 hours, collecting plankton in water at multiple points in lagoons, surrounding reefs and other sea areas by using a 300-mesh silk net, and filtering large individual impurities by using a large mesh; after fixing with alcohol, observing the number of larvae; after 7d of in-situ induced spawning, detecting the abundance and the number of the larvae of the instep tray again; in order to determine whether giant clam larvae are giant clam offspring or not in-situ spawning offspring, the giant clam larvae need to be identified by specific molecular markers.
Preferably, in the step d, a plurality of individuals are retrieved for paternity test, the in-situ induced spawning efficiency is detected, the quantity of the offspring obtained by induced spawning is calculated by retrieving the plurality of individuals for paternity test, DNA of the induced spawning parents and the field offspring seeds is used as a material, the in-situ induced spawning efficiency is detected by using markers such as microsatellites and the like, and the quantity of the offspring obtained by induced spawning is calculated.
The method is different from a fully artificial breeding method of giant clams, the field semi-closed lagoons and other environments are screened as spawning inducing fields, in-situ spawning induction is carried out at night, the production process of artificial seedlings is simulated as much as possible, risks and enemies are avoided, and artificial spawning inducing type seedlings consistent with wild seedlings are obtained in batches; and (3) carrying out paternity recognition by combining a modern molecular marker technology, and evaluating the in-situ induced spawning effect. According to the giant clam larva breeding method, the floating characteristics of giant clams in the larva stage are fully utilized, large-area diffusion is realized by means of sea waves, ocean currents and the like, a part of giant clam larva gathering areas are formed, sufficient parents are provided for giant clam breeding in future, and technical support is provided for quick repair of giant clam resources in south China sea. The invention has the advantages of simple and easy operation, obvious effect and the like.
The specific implementation mode is as follows:
the following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1:
a. parent ripening: in 3 middle of 2020 th month, in the 10m water depth non-wave zone environment of the lagoon Yongxing island lagoon in Xisha Taoist in China, 2 divers investigate giant clam resources and find 13 adult Nowa giant clams (with the shell length of 20-30cm) distributed in 3m2Within the area of (2), the station is positioned by using the GPS; through visual observation, the gonad part is found to be orange red, which indicates that the gonad is not mature, a little glandular part tissue is extracted by using an injector, and a small part of sperms can jump, the diameter of the ova is 80-90 mu m, and the ova are not mature; until 5 months and 1 day, the site is found through GPS positioning, the gonads of the giant clams are found to be brownish yellow after diving, which indicates that the giant clams are mature, and further microscope detection is adopted, so that the giant clams are found to be active in sperms, the diameter of each ovum is 100 micrometers, the color of each yolk is dark, and the giant clams indicate that the ovum is mature;
b. in-situ induced spawning: after the gonads of the giant clam are matured, a diver dives at 8 o' clock at night with a waterproof flashlight to dive at night, after giant clams are found, a serotonin solution is injected into mantle of 10 giant clam according to the dose of 0.3mg/kg by using an injector, 2-3min later, the giant clam begins to discharge sperms, lasts for about 15min, stops for about 10min, and begins to discharge eggs; the diver collects eggs with a 400-mesh bag prepared in advance; then, the other 3 giant clams are induced to spawn, when sperms are sprayed and put in the period, the collected ova are scattered in the vaporific sperms, the fertilization effect is realized, and then fertilized ova flow along with sea waves; the whole induced spawning process lasts for about 60min, and then divers return to the side of the lagoons;
c. larva examination: at 8 nights in 2 days after 5 months and 2 days, a diver collects plankton in water in lagoons by using a 300-mesh silk net, then filters by using a 100-mesh silk net, and collects the plankton which is fixed by 95% alcohol; then brought back to the laboratory of the western sand ocean observation station of Chinese academy of sciences, and a certain number of D-shaped larvae with the density of about 25 larvae per m in lagoons are discovered by using a microscope3The water depth of lagoons is estimated to be 1000 mu x 667 square meters x 3 metersAbout 5002.5 ten thousand D-shaped larvae exist in the whole process as 200.1 ten thousand water bodies; in order to determine whether the larvae are giant clam larvae or induced spawning larvae, molecular identification is carried out (see Ma, H., Feng, H., Zhang, Y., Qin, Y., Xiaoing, Z., Li, J., Zhang, Y., Yu, Z. multiple species-specific PCR identification of natural parent larvae in the South China Sea: a useful toy for application in the field of clinical stock management and sensory identification. Aquacucu 2021,531: 735991.), and about 50% of the larvae in the batch are larvae and are offspring of the parents, namely 2500 ten thousand larvae are obtained by the in-situ induction; in the morning of 5 months and 9 days, a diver collects plankton in the lagoons by using a 200-mesh silk net, then filters the plankton in the lagoons by using a 80-mesh silk net, and the collected plankton is fixed by using 95% alcohol; then brought back to the laboratory of the western sand ocean observation station of Chinese academy of sciences, and a certain number of face plate larvae with the density of 3/m are found in lagoons by using a microscope3According to the estimation of water bodies of lagoons, about 600 million podium disc larvae exist in the whole process; in order to determine whether the larvae are giant clam larvae or induced spawning larvae, molecular identification is carried out, and it is proved that about 30% of larvae in the batch are larvae in the batch and are offspring of the parents, that is to say, 180 ten thousand of instep pan larvae are obtained by in-situ induced spawning;
d. and (3) evaluating the spawning induction efficiency: checking whether newborn tridacna seedlings exist around lagoons in 12-10 days in 2020 by a diving mode, finding wild tridacna seedlings (the shell length is 10-15mm) by a section method, wherein the number of the wild tridacna seedlings is about 50 per hundred square meters, and estimating about 3.335 ten thousand of newborn tridacna seedlings according to the condition that 1000 mu multiplied 667 square meters are 66.7 ten thousand square meters; in order to determine whether the seedlings are obtained by in-situ induced spawning, molecular identification is carried out, and about 50% of larvae in the batch are filial generations of in-situ induced spawning parents, namely 16675 tridacna seedlings are obtained by the in-situ induced spawning.
The above results illustrate that: the norva tridacna parent in-situ induced spawning technology is effective and feasible, most importantly, the obtained offspring seeds are fixed on corallite and are drilled into a pit, so that the offspring seeds are well protected, and the in-situ induced spawning type offspring seeds are the same as wild offspring seeds and have better environmental adaptability. Most importantly, due to the influence of sea waves and ocean currents, the method for obtaining the giant clams forms a part of wild giant clams inhabitation area, provides sufficient parents for future reproduction and growth, and is beneficial to the rapid repair of giant clams in south China sea.
Example 2:
a. parent ripening: in the 3 rd month beginning of 2021 year, in the 10m water depth non-wave zone environment of Yoxing island lagoon in Xisha Islands in China, 30 tridacna parents with shells of 30-40cm are placed by 2 divers, and the tridacna parents are placed in 5m2Forming a thought giant clam parent group, firstly protecting the giant clam parent group by using a net cage, regularly observing the survival state, opening the net cage after completely adapting to the environment in the current period, and positioning the station by using a GPS (global positioning system); through visual observation, the gonad part is found to be orange red, which indicates that the gonad is not mature, a little glandular part tissue is extracted by using an injector, and a small part of sperms can jump, the diameter of the ova is 80-90 mu m, and the ova are not mature; until 5 months and 7 days, the site is found through GPS positioning, the gonads of the giant clams are found to be brownish yellow after diving, which indicates that the giant clams are mature, and further microscope detection is adopted, so that the giant clams are found to be active in sperms, the diameter of each ovum is 100 microns, the color of each yolk is dark, and the giant clams indicate that the ovum is mature;
b. in-situ induced spawning: after the gonads of the giant clams mature, a diver carries a waterproof flashlight to dive at 8 o' clock at night, after finding giant clams, the diver utilizes an injector to inject a serotonin solution into mantle of 25 giant clams according to a dose of 0.5mg/kg, after 3-5min, the giant clams begin to discharge sperms, the duration lasts for about 12min, about 8min is stopped, and the ova begin to be discharged; the diver collects eggs with a 400-mesh bag prepared in advance; then, hastening parturition on the other 5 giant clams, scattering the collected eggs in fog-like sperms when a large number of sperms are sprayed in the period, so that fertilization is realized, and then fertilized eggs flow along with sea waves; the whole induced spawning process lasts for about 60min, and then divers return to the side of the lagoons;
c. larva examination: at 8 nights in 5 months and 8 days, divers collect plankton in lagoons by using 300-mesh silk net, and then benefitFiltering with 100 mesh silk net, collecting organisms, and fixing with 95% ethanol; then brought back to the laboratory of the western sand ocean observation station of Chinese academy of sciences, and a certain number of D-shaped larvae with the density of about 60 larvae per m are found in lagoons3According to the water depth of lagoons, 1000 mu multiplied by 667 square meters multiplied by 3 meters is estimated to be 200.1 ten thousand of water, and about 1.2 hundred million D-shaped larvae exist in the whole process; in order to determine whether the larvae are giant clam larvae or induced spawning larvae, molecular identification is carried out (see Ma, H., Feng, H., Zhang, Y., Qin, Y., Xiaoing, Z., Li, J., Zhang, Y., Yu, Z. multiple species-specific PCR identification of natural parent larvae in the South China Sea: a useful toy toxin for application in the parent livestock storage and sensory identification. Aquacucu 2021,531: 735991), and about 60% of the larvae in the batch are larvae and are offspring of the parents, namely, 7200 larvae are obtained by the in situ induction; in the morning of 5 months and 15 days, a diver collects plankton in water in lagoons by using a 200-mesh silk net, then filters the plankton in the lagoons by using a 80-mesh silk net, and the collected plankton is fixed by using 95% alcohol; then brought back to the laboratory of the western sand ocean observation station of Chinese academy of sciences, and a certain number of face plate larvae with the density of about 8/m are found in lagoons by using a microscope3According to the estimation of water bodies of lagoons, about 1600-thousand-footed face plate larvae exist in the whole process; in order to determine whether the larvae are giant clam larvae or not and whether the larvae are induced spawning larvae or not, molecular identification is carried out, and it is proved that about 50% of the larvae in the batch are larvae in the batch and are offspring of the parents, namely 800 ten thousand of instep pan larvae are obtained by in-situ induced spawning;
d. and (3) evaluating the spawning induction efficiency: in 2021, 10 and 20 days, checking whether newborn tridacna seedlings are around lagoons through a diving mode, finding wild tridacna seedlings (the shell length is 10-30mm) by a section method, wherein the number of the wild tridacna seedlings is about 20 per hundred square meters, and estimating about 13.34 ten thousand of the newborn tridacna seedlings according to the condition that 1000 mu of the wild tridacna seedlings are 66.7 ten thousand square meters; in order to determine whether the seedlings are obtained by in-situ induced spawning, molecular identification is carried out, and it is proved that about 45% of larvae of the batch are offspring of in-situ induced spawning parents, namely about 6 ten thousand tridacna seedlings are obtained by the in-situ induced spawning.
The above results illustrate that: the in-situ spawning induction technology of tridacna parents is effective and feasible, the most important thing is that the obtained fries are fixed on corallite and are drilled into a pit, so that the tridacna parents can well protect the tridacna parents, and the in-situ spawning induction type fries are the same as wild fries and have better environmental adaptability. Most importantly, due to the influence of sea waves and ocean currents, the method for obtaining the giant clams forms a part of wild giant clams inhabitation area, provides sufficient parents for future reproduction and growth, and is beneficial to the rapid repair of giant clams in south China sea.
Claims (9)
1. An in-situ spawning induction method of tridacna parents is characterized by comprising the following steps:
a. gonad monitoring: finding a distribution group of wild or artificially transplanted giant clams by diving in a 5-30 m water-depth wave-free area environment of a south-sea field island reef sea area, a lagoon or a ring reef in China, positioning, recording the site position of a parent group, and regularly observing the gonad development condition;
b. in-situ induced spawning: when the gonads mature, diving to a giant clam group distribution area, and injecting an oxytocic into a mantle for in-situ oxytocic;
c. larva examination: after the in-situ spawning of the parents is promoted, plankton in water is collected at multiple points in the island reef sea area, the lagoons or the ring reef sea area, and large individual impurities are filtered; after fixing with alcohol, observing the number of larvae; then detecting the abundance and the number of the podosphaeria pan larvae; identifying whether giant clam larvae are giant clam offspring or not in-situ induced spawning offspring;
d. and (3) evaluating the spawning induction efficiency: and after 6-9 months of induced spawning, checking whether newly born giant clam seedlings of about 10-30mm are around island reef sea areas, lagoons and surrounding reefs where the in-situ induced spawning sites are located, taking back a plurality of individuals, performing paternity test, detecting the in-situ induced spawning efficiency, and calculating the number of offspring obtained by induced spawning.
2. The in-situ spawning method of tridacna sp parent according to claim 1, wherein in the step b, the spawning agent is a serotonin solution, and the dosage of the spawning agent is within the range of 0.3-0.5mg/kg of total weight of serotonin/tridacna sp.
3. The in-situ spawning induction method of tridacna sp parent according to claim 1, characterized in that in the step c, the specific molecular marker is a composite PCR primer combination capable of distinguishing more than 3-5 species of tridacna sp at the same time, so that larva compositions and parent sources thereof can be effectively distinguished.
4. The in-situ spawning induction method of tridacna sp parent according to claim 1, characterized in that in the step d, the feasibility of in-situ spawning induction efficiency extremely recovering resources is evaluated through the number and variety of tridacna sp per unit area and a parent-offspring recognition technology.
5. The in-situ spawning induction method of tridacna sp parent according to claim 1, wherein the step a is performed by positioning with a GPS.
6. The in-situ spawning induction method of tridacna sp parent according to claim 1, wherein the step a of periodically observing gonad development is to periodically observe gonad development by a method combining visual observation and microscopic sampling.
7. The in-situ spawning promotion method of tridacna parents according to claim 1, characterized in that in the step b, only one tridacna is selected for injection each time during in-situ spawning promotion; the spawning induction time is controlled within 1 h; the spawning induction time is 8-10 o' clock at night; collecting the discharged eggs, then dispersing and combining with newly discharged sperms to promote the fertilized eggs to drift along with sea waves.
8. The method for in-situ spawning induction of tridacna sp parent according to claim 1, wherein the larva examination in step c is as follows: after the parent is induced to spawn in situ for 24 hours, collecting plankton in water at multiple points in lagoons, surrounding reefs and other sea areas by using a 300-mesh silk net, and filtering large individual impurities by using a large mesh; after fixing with alcohol, observing the number of larvae; after 7d of in-situ induced spawning, detecting the abundance and the number of the larvae of the instep tray again; in order to determine whether giant clam larvae are giant clam offspring or not in-situ spawning offspring, the giant clam larvae need to be identified by specific molecular markers.
9. The in-situ spawning induction method of tridacna sp parent according to claim 1, wherein in the step d, a plurality of individuals are retrieved for paternity test to detect the in-situ spawning induction efficiency, the quantity of the spawns obtained by calculating the induced spawning is retrieved for a plurality of individuals for paternity test, the DNA of the induced spawning parent and the field offspring seeds is used as a material, the in-situ spawning induction efficiency is detected by using markers such as microsatellites and the like, and the quantity of the spawns obtained by inducing spawning is calculated.
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