CN116784264A - Induction method of total-female salmon strain - Google Patents
Induction method of total-female salmon strain Download PDFInfo
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Classifications
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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
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0278—Physical preservation processes
-
- 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)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Biophysics (AREA)
- Physiology (AREA)
- Marine Sciences & Fisheries (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention relates to the technical field of salmon strain induction, in particular to a method for inducing a full-female salmon strain, which comprises the following steps: cultivating and hastening parthenogenesis of pseudo-male parent fish, purchasing XX-type full-female eye spawn, hatching into a larval fish opening, then feeding androgens, inducing to obtain XX-type pseudo-male fish, screening sexual precocity individuals through molecular markers, and carrying out environment regulation and induction sexual maturity to obtain sperms; step two: female parent fish is cultivated and induced spawning is carried out, physiological female fish with age of more than 2 is purchased, after nutrition is enhanced for 6-8 months, the female parent fish is induced to mature through environmental regulation and control, ovum is obtained through artificial induced spawning, and fertilized ovum is prepared through artificial insemination; step three: and (3) preparing the full-female offspring seeds, and incubating the fertilized eggs in the second step to obtain the full-female offspring seeds. According to the invention, under the premise of not influencing sperm quality, the pseudo-male fish can be mature in advance through molecular marker screening and environmental regulation, so that the seed production period is shortened, and the seed production efficiency is improved.
Description
Technical Field
The invention relates to the technical field of salmon strain induction, in particular to a method for inducing a full-female salmon strain.
Background
Atlantic salmon, commonly known as salmon, is a worldwide farmed fish, cultivated in tens of countries worldwide, and is a major impetus for the growth of global fishery and aquaculture trade in recent decades. In almost all areas, the demand for atlantic salmon increases faster than other fish. Atlantic salmon cultivation by using various cold water resources (underground cold water, cold water clusters, power station cooling water and the like) is started and succeeded in China, and the deep-sea cultivation of the Atlantic salmon by using a deep-sea net cage in China is also successful at present, so that along with the improvement of the quantity of cultivated bodies in China, the demand of offspring seed is greatly increased, the self-sufficient call of the offspring seed of the Atlantic salmon in the industry is higher and higher, and the localization of offspring seed is imperative.
Atlantic salmon (salmon) is one of the main sea water consumption varieties in China, and a plurality of deep sea net cages and land-based industrial culture Atlantic salmon projects are developed in China at present, so that a full-female salmon strain induction method is needed.
Disclosure of Invention
The invention aims to provide a method for inducing a full-female salmon strain so as to solve the problems in the background technology.
In order to achieve the above purpose, the present invention provides the following technical solutions: a method for inducing a full-female salmon strain, comprising the following steps:
step one: cultivating and hastening parturition of pseudo-male parent fish, purchasing XX-type full-female eye spawn, feeding androgens after hatching into a larval fish opening, inducing to obtain XX-type pseudo-male fish, then silvering the pseudo-male fish individuals into sea salmon, cultivating the sea salmon to 2 years old, screening sexual precocity individuals by using molecular markers, inducing the pseudo-male fish to ripen through environmental regulation, artificially hastening parturition to obtain sperms for insemination or ultralow temperature freezing preservation, and cultivating the pseudo-female fish incapable of precocity to natural maturity;
step two: female parent fish is cultivated and induced spawning is carried out, physiological female fish with age of more than 2 is purchased, after nutrition is enriched for 6-8 months, the female parent fish is induced to mature through environmental regulation and control, ovum is obtained through artificial induced spawning, and fertilized ovum is obtained through artificial insemination by using fresh sperm or frozen sperm;
step three: and (3) preparing the full-female offspring seeds, incubating the fertilized eggs in the second step for 6 months, checking gonad development, and determining female proportion to obtain the full-female offspring seeds.
Preferably, the specific steps of breeding and spawning induction of the pseudo-male parent fish are as follows:
(1) Purchasing a full female diploid ophthalmonogy egg, and incubating to obtain XX-type larva fish;
(2) Feeding 17 alpha-methyltestosterone to induce reverse rotation after feeding artificial compound feed to the opening of the fish fry to obtain XX-type pseudo-male fish; silver treatment is carried out when the fish fry grows to 20 g, and 100g of the young fish is transferred into sea water for cultivation;
(3) Checking gonad morphology, determining induction effect, simplifying genome sequencing or genome resequencing for pseudo-male fish with normal development of testis, pseudo-male fish with dysgenesis of testis and intermittent individuals, developing molecular markers related to successful induction character, and selecting pseudo-female fish for production;
(4) After the selected pseudo-female fish is cultured in the sea water for 12-14 months, the gonadal development is checked periodically during the culture period, when GSI is more than 0.3 and a large number of sperm cells are observed in a tissue section, the developed molecular markers are utilized to screen individuals which can mature in advance, the pseudo-male fish is cultured in fresh water, the light control temperature is controlled to promote sexual maturation, and the rest pseudo-male fish is continuously cultured in the sea water;
(5) 7-14 days after the second sex characteristics of the pseudo-male fish cultivated in the fresh water appear, carrying out artificial induced spawning by utilizing LHRHA, carrying out anesthesia by using MS-222 before induced spawning, and carrying out injection metering at 500 mu l/kg, wherein the consumption of the LHRHA is 5 mu g/kg, and carrying out back intramuscular injection;
(6) After spawning is induced for 4-7 days, killing male, taking sperms, cutting spermary, filtering with gauze, taking a small amount of semen, activating and microscopic examination, and then using the semen for artificial insemination or preserving the semen in a liquid nitrogen tank for later use;
(7) And (3) continuously culturing the rest non-desalted cultured pseudo-male fishes in seawater, periodically checking gonad development condition in the culturing process, desalting and culturing the pseudo-male fishes after culturing for 6-12 months, controlling light temperature to promote sexual maturity, and repeating the operations of (5) and (6).
Preferably, the specific method for cultivating and hastening parturition of female parent fish comprises the following steps:
(1) Purchasing physiological female fishes with ages of more than 2 in different batches, and carrying out nutrition enhancement cultivation on the physiological female fishes by utilizing fish oil or astaxanthin, wherein the crude protein content of the feed is more than or equal to 42%, and the daily feeding amount is 1% of the weight;
(2) After the female fish is subjected to operation and culture reinforcement for 6-8 months, checking gonadal development, culturing the female fish into fresh water when GSI is more than 0.6% and the egg diameter is more than 5 mm, and controlling light control temperature to promote sexual maturity;
(3) 7-14 days after the female parent fish has the second sex characteristics, utilizing LHRHA to carry out artificial induced spawning, and carrying out anesthesia by MS-222 before induced spawning;
(4) After spawning induction for 4-7 days, checking spawning conditions of female parent fishes, picking spawnable parent fishes in time for artificial insemination, checking 1 time every 7 days, and selecting spawnable parent fishes, wherein spawning induction groups can complete spawning within 1 month;
(5) And (3) continuously culturing the rest non-desalted female fishes in seawater, periodically checking gonad development condition in the culturing process, desalting and culturing the female fishes after culturing for 6-12 months, controlling light temperature to promote sexual maturity, and repeating the operations of (3) and (4).
Preferably, the specific method for preparing the total female offspring seed comprises the following steps:
(1) Dry insemination, wherein the ratio of male to female is 2:1, and hatching in an incubator after insemination, cleaning and disinfection;
(2) Hatching fertilized eggs, breeding young fish and silvering young fish;
(3) Counting the sex proportion of the salmon with age of more than 6 months, and determining the female proportion.
Preferably, the induction amount of 17 alpha-methyltestosterone is 5mg/kg, and the induction time is 900 days; the water temperature of the juvenile fish transferred into mariculture is not more than 18 ℃, preferably 14-16 ℃, and the illumination is 1000lux and 24L:0D.
Preferably, the desalination method for the pseudo-male fish dilution comprises the following steps: the salinity is reduced by 3 per mill to 5 per mill every 3 days, the water temperature is reduced by 1 ℃ to below 10 ℃ every day before desalination, preferably 5 to 8 ℃ and the illumination is 200 to 500lux and 12L:12D.
Preferably, vitamin E and ascorbic acid are added into the sperm preservation solution to improve sperm motility.
Preferably, when the female parent fish has secondary sex characteristics for induced spawning, back intramuscular injection is adopted, the induction of spawning injection is metered to be 500 mu l/kg, and the LHRHA dosage is 10 mu g/kg.
Compared with the prior art, the invention has the beneficial effects that: the method for inducing the full-female salmon strain prepares the full-female offspring seeds of the Atlantic salmon by utilizing the methods of sex reversal technology, molecular marking, environmental regulation, ultralow temperature cryopreservation of sperms and the like, and has high seed production efficiency and short seed production period; screening related molecular markers of the induced success character (sex reversal and normal development of spermary), establishing a quick detection method of the pseudo-male fish, screening the pseudo-male fish meeting production requirements for cultivation, and saving cultivation cost; moreover, aiming at the phenomena that the Atlantic salmon pseudo-male fish cannot discharge sperms, the male and female sexual maturation time is asynchronous and the female fish spawning time is long, the ultra-low temperature freezing technology is utilized to store the sperm of the pseudo-male fish, the activity of the sperm is stored for a long time, and the utilization rate of the pseudo-male fish is improved; the parent fish cultivation, fertilized egg incubation, offspring seed cultivation and other processes are all carried out in a circulating water system, and environmental factors such as temperature, illumination, salinity and the like can be accurately controlled, so that optimal conditions are ensured; the technique for producing the Atlantic salmon total female offspring seeds can realize annual production without season limitation, and meets offspring seed requirements of different areas and different times in China.
Drawings
FIG. 1 is a flow chart of the present invention.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and all other embodiments obtained by those skilled in the art without making creative efforts based on the embodiments of the present invention are included in the protection scope of the present invention.
The structure of the induction method of the total-female salmon strain provided by the invention is shown in figure 1, and the induction method comprises the following steps:
step one: culturing and hastening parthenogenesis of pseudo-male parent fish, purchasing XX-type full-female eye spawn, feeding androgens after hatching into a larval fish opening, inducing to obtain XX-type pseudo-male fish, then silver-forming the pseudo-male individual into sea salmon, culturing in sea water to 2 years old, screening sexual precocity individual by molecular markers, inducing the pseudo-male fish to mature through environmental regulation (salinity reduction, light control and temperature control), artificially hastening parthenogenesis to obtain sperms for insemination or ultralow temperature freezing preservation, and culturing the pseudo-female fish incapable of precocity to natural maturity;
further, the specific steps of breeding and spawning induction of the pseudo-male parent fish are as follows:
(1) Purchasing a full female diploid (XX) ophthalmonogy egg, and incubating to obtain an XX-type larva fish;
(2) Feeding 17 alpha-methyltestosterone (17-alpha MT) after feeding artificial compound feed to the opening of the fish fry, and performing induced inversion to obtain XX-type pseudo-male fish, wherein the induction measurement is 5mg/kg, and the induction time is 900 days; silvering when the fish fry grows to 20 g, transferring the young fish into sea water for cultivation when 100g, and culturing at the water temperature of 14-16 ℃ under the illumination of 1000lux and 24L:0D;
(3) Checking gonad morphology, determining induction effect, simplifying genome sequencing or genome resequencing for pseudo-male fish with normal development of spermary, pseudo-male fish with abnormal development of spermary and intermittent individuals, developing molecular markers related to successful induction character (sex reversal and normal development of gonad), and selecting pseudo-female fish for production;
(4) After the selected pseudo-female fish is cultured in the sea water for 12-14 months, the gonadal development is checked periodically during the culture period, when GSI is more than 0.3 and a large number of sperm cells are observed in a tissue section, the developed molecular markers are utilized to screen individuals which can mature in advance, the pseudo-male fish is cultured in fresh water, the light control temperature is controlled to promote sexual maturation, the rest pseudo-male fish is continuously cultured in the sea water, and the method for desalting the pseudo-male fish comprises the following steps: the salinity is reduced by 3 per mill to 5 per mill every 3 days, the water temperature is reduced by 1 ℃ to below 10 ℃ every day before desalination, preferably 5 to 8 ℃ and the illumination is 200 to 500lux and 12L:12D;
(5) 7-14 days after the second sex characteristics of the pseudo-male fish cultivated in the fresh water appear, carrying out artificial induced spawning by utilizing LHRHA, carrying out anesthesia by using MS-222 (50-100 ppm) before induced spawning, measuring 500 mu l/kg by injection, and carrying out back intramuscular injection by using 5 mu g/kg of LHRHA;
(6) After 4-7 days of induced spawning, killing male, taking sperms, cutting sperms, filtering with gauze, taking a small amount of semen, performing activation microscopic examination, and using the semen for artificial insemination or preserving the semen in a liquid nitrogen tank for later use, wherein vitamin E (0.1 mM) and ascorbic acid (1 mM) are added into the semen preserving liquid, so that the sperm motility is improved;
(7) Continuously culturing the rest non-desalted cultured pseudo-male fish in sea water, periodically checking gonad development condition in the culturing process, desalting and culturing the pseudo-male fish after culturing for 6-12 months, controlling light control temperature to promote sexual maturity, and repeating the operations of (5) and (6);
step two: female parent fish is cultivated and induced spawning, physiological female fish with age of more than 2 is purchased, nutrition is enriched for 6-8 months, and then induced sexual maturity is achieved through environmental regulation (salinity reduction, light control and temperature control), ovum is obtained through artificial induced spawning, and fertilized ovum is obtained through artificial insemination by utilizing fresh sperm or frozen sperm.
Further, the specific method for cultivating and hastening parturition of female parent fish comprises the following steps:
(1) Purchasing physiological female fishes with ages of more than 2 in different batches, and carrying out nutrition enhancement cultivation on the physiological female fishes by utilizing fish oil or astaxanthin, wherein the crude protein content of the feed is more than or equal to 42%, and the daily feeding amount is 1% of the weight; the water temperature is 14-16 ℃, and the illumination is 1000lx and 24L to 0D;
(2) After the female fish is enriched for 6-8 months, checking gonadal development, and when GSI is more than 0.6% and the egg diameter is more than 5 mm, promoting the female fish to mature in fresh water, and the method for desalting the female fish comprises the following steps: the salinity is reduced by 3 per mill to 5 per mill every 3 days, the water temperature is reduced by 1 ℃ to below 10 ℃ every day before desalination, preferably 5 to 8 ℃ and the illumination is 200 to 500lux and 12L:12D;
(3) 7-14 days after the female parent fish has the second sex characteristics, carrying out artificial induced spawning by utilizing LHRHA, and carrying out anesthesia by using MS-222 (50-100 ppm) before induced spawning, wherein the injection quantity for induced spawning is 500 mu l/kg, the dosage of LHRHA is 10 mu g/kg, and the injection is carried out on back muscle;
(4) After spawning induction for 4-7 days, checking spawning conditions of female parent fishes, picking spawnable parent fishes in time for artificial insemination, checking 1 time every 7 days, and selecting spawnable parent fishes, wherein spawning induction groups can complete spawning within 1 month;
(5) Continuously culturing the rest non-desalted female fishes in sea water, checking gonad development condition periodically in the culturing process, desalting and culturing the female fishes after culturing for 6-12 months, controlling light temperature to promote sexual maturity, and repeating the operations of (3) and (4);
step three: and (3) preparing the full-female offspring seeds, incubating the fertilized eggs in the second step for 6 months, checking gonad development, and determining female proportion to obtain the full-female offspring seeds.
Further, the specific method for preparing the full female offspring seed comprises the following steps:
(1) Dry insemination, wherein the ratio of male to female is 2:1, and hatching in an incubator after insemination, cleaning and disinfection;
(2) Hatching fertilized eggs, breeding young fish and silvering young fish;
(3) Counting the sex proportion of the salmon with age of more than 6 months, and determining the female proportion.
The invention prepares the Atlantic salmon total female offspring seeds by utilizing the methods of sex reversal technology, molecular marking, environmental regulation, sperm ultralow temperature cryopreservation and the like, and has high seed production efficiency and short seed production period; screening related molecular markers of the induced success character (sex reversal and normal development of spermary), establishing a quick detection method of the pseudo-male fish, screening the pseudo-male fish meeting production requirements for cultivation, and saving cultivation cost; moreover, aiming at the phenomena that the Atlantic salmon pseudo-male fish cannot discharge sperms, the male and female sexual maturation time is asynchronous and the female fish spawning time is long, the ultra-low temperature freezing technology is utilized to store the sperm of the pseudo-male fish, the activity of the sperm is stored for a long time, and the utilization rate of the pseudo-male fish is improved; the parent fish cultivation, fertilized egg incubation, offspring seed cultivation and other processes are all carried out in a circulating water system, and environmental factors such as temperature, illumination, salinity and the like can be accurately controlled, so that optimal conditions are ensured; the technique for producing the Atlantic salmon total female offspring seeds can realize annual production without season limitation, and meets offspring seed requirements of different areas and different times in China.
The method comprises the following steps: because the inherited sex type of Atlantic salmon is XX/XY type, the total male breeding technique method utilizes androgens to induce XX type female fish to sex reversal to XX type pseudo-male fish, the XX type pseudo-male fish is hybridized with XX type female fish to obtain XX type total female offspring, individual difference exists in induction effect of the pseudo-male fish, and molecular markers related to induction success characters (sex reversal and normal gonad development) can be developed to rapidly screen the pseudo-male fish meeting requirements for breeding.
In addition, sex maturation related molecular markers are used for screening sex early-maturing pseudo-male fishes, the sex maturation of the sex early-maturing pseudo-male fishes is promoted by environmental regulation, the seed production time is shortened, the female fishes generally need to be sexually mature of more than 3 years, the sex early-maturing can be regulated by the environment, but the premature maturation of small individual female fishes (such as 1 year) can cause poor ovum quality to influence the hatching rate and the survival rate of young fishes, therefore, the female fishes of more than 2 years old are selected as female parents for carrying out operation and cultivation to strengthen the female fishes to 3 years old, and the sperm is preserved by using an ultralow temperature freezing method aiming at the problems of asynchronous female male maturation time (male pre-maturing) and long spawning period of the female fishes.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (8)
1. The induction method of the total-female salmon strain is characterized by comprising the following steps of:
step one: cultivating and hastening parturition of pseudo-male parent fish, purchasing XX-type full-female eye spawn, feeding androgens after hatching into a larval fish opening, inducing to obtain XX-type pseudo-male fish, then silvering the pseudo-male fish individuals into sea salmon, cultivating the sea salmon to 2 years old, screening sexual precocity individuals by using molecular markers, inducing the pseudo-male fish to ripen through environmental regulation, artificially hastening parturition to obtain sperms for insemination or ultralow temperature freezing preservation, and cultivating the pseudo-female fish incapable of precocity to natural maturity;
step two: female parent fish is cultivated and induced spawning is carried out, physiological female fish with age of more than 2 is purchased, after nutrition is enriched for 6-8 months, the female parent fish is induced to mature through environmental regulation and control, ovum is obtained through artificial induced spawning, and fertilized ovum is obtained through artificial insemination by using fresh sperm or frozen sperm;
step three: and (3) preparing the full-female offspring seeds, incubating the fertilized eggs in the second step for 6 months, checking gonad development, and determining female proportion to obtain the full-female offspring seeds.
2. A method for inducing a strain of whole female salmon according to claim 1, wherein: in the first step, the specific steps of breeding and spawning induction of the pseudo-male parent fish are as follows:
(1) Purchasing a full female diploid ophthalmonogy egg, and incubating to obtain XX-type larva fish;
(2) Feeding 17 alpha-methyltestosterone to induce reverse rotation after feeding artificial compound feed to the opening of the fish fry to obtain XX-type pseudo-male fish; silver treatment is carried out when the fish fry grows to 20 g, and 100g of the young fish is transferred into sea water for cultivation;
(3) Checking gonad morphology, determining induction effect, simplifying genome sequencing or genome resequencing for pseudo-male fish with normal development of testis, pseudo-male fish with dysgenesis of testis and intermittent individuals, developing molecular markers related to successful induction character, and selecting pseudo-female fish for production;
(4) After the selected pseudo-female fish is cultured in the sea water for 12-14 months, the gonadal development is checked periodically during the culture period, when GSI is more than 0.3 and a large number of sperm cells are observed in a tissue section, the developed molecular markers are utilized to screen individuals which can mature in advance, the pseudo-male fish is cultured in fresh water, the light control temperature is controlled to promote sexual maturation, and the rest pseudo-male fish is continuously cultured in the sea water;
(5) 7-14 days after the second sex characteristics of the pseudo-male fish cultivated in the fresh water appear, carrying out artificial induced spawning by utilizing LHRHA, carrying out anesthesia by using MS-222 before induced spawning, and carrying out injection metering at 500 mu l/kg, wherein the consumption of the LHRHA is 5 mu g/kg, and carrying out back intramuscular injection;
(6) After spawning is induced for 4-7 days, killing male, taking sperms, cutting spermary, filtering with gauze, taking a small amount of semen, activating and microscopic examination, and then using the semen for artificial insemination or preserving the semen in a liquid nitrogen tank for later use;
(7) Continuously culturing the rest non-desalted cultured pseudo-male fish in seawater, periodically checking gonad development condition during culturing, culturing for 6-12 months, culturing the pseudo-male fish in fresh water, controlling light temperature to promote sexual maturity, and repeating the operations of (5) and (6).
3. A method for inducing a strain of whole female salmon according to claim 1, wherein: in the second step, the specific method for cultivating and hastening parturition of the female parent fish comprises the following steps:
(1) Purchasing physiological female fishes with ages of more than 2 in different batches, and carrying out nutrition enhancement cultivation on the physiological female fishes by utilizing fish oil or astaxanthin, wherein the crude protein content of the feed is more than or equal to 42%, and the daily feeding amount is 1% of the weight;
(2) After the female fish is subjected to operation and culture reinforcement for 6-8 months, checking gonadal development, culturing the female fish into fresh water when GSI is more than 0.6% and the egg diameter is more than 5 mm, and controlling light control temperature to promote sexual maturity;
(3) 7-14 days after the female parent fish has the second sex characteristics, utilizing LHRHA to carry out artificial induced spawning, and carrying out anesthesia by MS-222 before induced spawning;
(4) After spawning induction for 4-7 days, checking spawning conditions of female parent fishes, picking spawnable parent fishes in time for artificial insemination, checking 1 time every 7 days, and selecting spawnable parent fishes, wherein spawning induction groups can complete spawning within 1 month;
(5) And (3) continuously culturing the rest non-desalted cultured female fishes in seawater, checking gonad development condition periodically in the culturing process, controlling the temperature of the desalted female fishes to promote sexual maturity after culturing for 6-12 months, and repeating the operations of (3) and (4).
4. A method for inducing a strain of whole female salmon according to claim 1, wherein: in the third step, the specific method for preparing the total female offspring seed comprises the following steps:
(1) Dry insemination, wherein the ratio of male to female is 2:1, and hatching in an incubator after insemination, cleaning and disinfection;
(2) Hatching fertilized eggs, breeding young fish and silvering young fish;
(3) Counting the sex proportion of the salmon with age of more than 6 months, and determining the female proportion.
5. A method for inducing a strain of whole female salmon according to claim 2, wherein: the induction amount of 17 alpha-methyltestosterone is 5mg/kg, and the induction time is 900 days; the water temperature of the juvenile fish transferred into mariculture is not more than 18 ℃, preferably 14-16 ℃, and the illumination is 1000lux and 24L:0D.
6. A method for inducing a strain of whole female salmon according to claim 2, wherein: the desalination method for the pseudo-male fish dilution comprises the following steps: the salinity is reduced by 3 per mill to 5 per mill every 3 days, the water temperature is reduced by 1 ℃ to below 10 ℃ every day before desalination, preferably 5 to 8 ℃ and the illumination is 200 to 500lux and 12L:12D.
7. A method for inducing a strain of whole female salmon according to claim 2, wherein: vitamin E and ascorbic acid are added into the sperm preservation solution, so that sperm motility is improved.
8. A method of inducing a full-female salmon strain according to claim 3, wherein: when the female parent fish has secondary sex characteristics for induced spawning, back intramuscular injection is adopted, the induced spawning injection is metered to be 500 mu l/kg, and the LHRHA dosage is 10 mu g/kg.
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| CN110771536A (en) * | 2019-11-21 | 2020-02-11 | 浙江省海洋水产研究所 | Large-scale cultivation method of all-female spotted maigre |
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