Method for breeding seedling of jinhu hybrid spot

Abstract

The application provides a breeding method of golden tiger hybrid striped bass fry, which comprises the following steps: selecting a breeding environment; cultivating and maturing female parents; controlling before spawning; insemination; hatching of fertilized eggs; and fry breeding. In the step of cultivating and maturing female parents, specific biological bait is fed in stages to promote the feeding of parent fish, accelerate the growth, create favorable conditions for subsequent spawning by using specific mixed feed in the step of controlling before spawning, improve the quality and quantity of eggs, introduce low-frequency ultrasonic assisted fertilization in the step of artificial insemination to significantly improve the activity of eggs and sperm and improve the fertilization rate, supplement intermittent ultrasonic waves in the step of hatching of fertilized eggs to improve the hatching rate and the yield of golden tiger hybrid striped bass fry, and release beneficial bacteria in specific breeding ponds after hatching to improve the adaptability of fry, adsorb harmful substances, reduce pathogenic bacteria, stabilize water quality, promote the growth of fry, improve the survival rate, and reduce the rate of deformity.

Description

Technical Field

[0001] This invention relates to the field of aquaculture technology, and in particular to a method for cultivating seedlings of the Golden Tiger Hybrid. Background Technology

[0002] Grouper is an important marine fish germplasm resource in my country, playing a pivotal role in the industry, with an annual demand of 1 billion fry. However, the prevalence of diseases and low survival rates among grouper fry have become bottlenecks for the industry's development. By developing grouper breed improvement technologies and cultivating fast-growing, resilient, and superior germplasm, the problems of disease prevalence and low survival rates can be solved. The "Golden Tiger Hybrid Grouper" is a hybrid breed obtained by breeding a brown-spotted grouper as the mother and a blue-bodied large-spotted grouper as the father. It features rapid growth, a smaller coefficient of variation in body length and weight, and stable heritability of traits. The Golden Tiger Hybrid Grouper will provide a high-quality, suitable species for modern factory farming and deep-sea aquaculture in my country, and will strongly promote innovation in marine fisheries seed industry and sustainable development.

[0003] Currently, the artificial breeding technology for this hybrid variety is not yet mature. The parent fish have difficulty laying eggs, the quality of sperm and eggs is poor, the hatching rate of fertilized eggs is low, the survival rate of fry is low, some fry are deformed, and the disease resistance is poor. This has a certain impact on the yield and quality of golden tiger hybrid grouper fry, and greatly restricts the large-scale breeding process of large grouper.

[0004] Therefore, it is necessary to further optimize the grouper fry breeding process to improve the yield and quality of golden tiger hybrid grouper fry, promote the healthy development of the grouper industry, and advance the large-scale breeding of grouper, which is of practical significance. Summary of the Invention

[0005] Therefore, this invention proposes a method for cultivating seedlings of the Golden Tiger hybrid.

[0006] The technical solution of this invention is implemented as follows:

[0007] The seedling cultivation method for Golden Tiger Hybrid Spotted Varieties includes the following steps:

[0008] S1. Optimize the breeding environment: Select a pond with a water temperature of 22-25℃ and a water depth of 2-3 meters, and release female parent brown-spotted grouper.

[0009] S2. Female parent breeding and maturation: In the first stage, feed biological feed A at a rate of 4%-6% of the female parent's body weight, 3-4 times a day for 4-6 days. In the second stage, feed a mixture of biological feed B and nutritional preparations at a rate of 5%-7% of the female parent's body weight, 2-3 times a day for 3-5 days.

[0010] S3. Pre-spawning management: 5-7 days before the female parent lays eggs, raise the water temperature to 27-28℃ and feed them with formulated feed at a rate of 1%-3% of the female parent's body weight, twice a day.

[0011] S4. Fertilization: After the female parent lays eggs, the eggs are fertilized with sperm from the blue-bodied large-spotted grouper under low-frequency ultrasonic field conditions to obtain fertilized eggs.

[0012] S5. Fertilized egg hatching: Fertilized eggs are placed in an incubator and subjected to intermittent ultrasonic treatment to hatch into fish fry;

[0013] S6. Fish fry rearing: Select a rearing pond, add probiotic magnetic chitosan microspheres, and transfer the fish fry to the rearing pond for initial rearing.

[0014] Furthermore, in step S1, the female parent brown-spotted grouper is selected from healthy brown-spotted groupers that are over 6 years old and free from disease, with a stocking density of 100-200 fish / m². 2 .

[0015] Furthermore, in step S2, the first stage feeds biological bait A, which, by weight, includes the following ingredients: 30-40 parts fishmeal, 5-10 parts rotifers, 1-3 parts vitamins, 1-3 parts amino acids, 4-6 parts green fruit extract, 1-3 parts nightshade polysaccharide, 4-6 parts Dendrobium nobile stem extract, and 2-4 parts coconut oil.

[0016] Furthermore, the *Cephalotaxus fortunei* extract is a water extract and / or an ethanol extract of *Cephalotaxus fortunei*; the *Dendrobium nobile* stem extract is an ethanol extract and / or an ethyl acetate extract of *Dendrobium nobile* stem.

[0017] Furthermore, in step S2, the second stage of feeding biological feed B includes the following ingredients by weight: 20-30 parts fish meal, 4-6 parts shrimp meal, 1-3 parts vitamins, 1-3 parts tea oil polysaccharide, 1-3 parts shiitake mushroom polysaccharide, 1-2 parts Dioscorea bulbifera fruit extract, 3-5 parts bamboo orchid extract, and 2-4 parts blended vegetable oil.

[0018] Furthermore, the Dioscorea opposita fruit extract is a water extract and / or an ethanol extract of Dioscorea opposita fruit; the Oryza sativa extract is a water extract and / or an ethanol extract of Oryza sativa.

[0019] Furthermore, in step S2, the nutrient preparation comprises, by weight, the following raw materials: 3-5 parts earthworm fermentation liquid and 3-5 parts chitin fermentation liquid.

[0020] The preparation method of the nutrient preparation is as follows: earthworm fermentation liquid and chitin fermentation liquid are treated twice by a dynamic high-pressure micro-jet homogenizer at 80-120MPa, and then mixed to obtain the nutrient preparation.

[0021] Furthermore, the earthworm fermentation liquid is prepared by pulverizing earthworms and then ultrasonically stirring with lipase; the chitin fermentation liquid is prepared by crushing shrimp shells and then fermenting them with Trichoderma harzianum and Bacillus spp., with the preferred mass ratio of Trichoderma harzianum to Bacillus spp. being 1:1, and the preferred fermentation temperature being 35-45℃.

[0022] Furthermore, in step S3, the compound feed comprises, by weight, the following ingredients: 10-20 parts shrimp powder, 5-8 parts clam meat paste, 2-4 parts oxalic acid nutrient solution, 3-4 parts wild Ganoderma lucidum extract, 1-3 parts perilla leaf extract, 1-3 parts Morinda officinalis fruit extract, and 1-3 parts longan polysaccharide.

[0023] Furthermore, the wild Ganoderma lucidum extract is a wild Ganoderma lucidum aqueous extract and / or wild Ganoderma lucidum ethanol extract; the perilla leaf extract is a perilla leaf aqueous extract and / or perilla leaf ethanol extract; and the Morinda officinalis fruit extract is a Morinda officinalis fruit ethanol extract and / or Morinda officinalis fruit ethyl acetate extract.

[0024] Furthermore, in step S4, the low-frequency ultrasound field processing specifically involves: performing ultrasound at 20-30 kHz and 10-15 W for 20-30 seconds, followed by a 30-second interval, and then continuing ultrasound for another 20-30 seconds. This process is repeated until fertilization is completed.

[0025] Furthermore, in step S5, the intermittent ultrasound treatment involves first ultrasounding at 20-30 kHz and 20-30 W for 30-60 seconds, followed by a 3-minute interval, then ultrasounding at 50-60 kHz and 50-60 W for 20-40 seconds, followed by a 5-minute interval, and repeating the above process 3-5 times.

[0026] Further, in step S6, the preparation method of the probiotic magnetic chitosan microspheres includes: adding chitosan to water, transferring it to a high-pressure reactor, carrying out a hydrothermal reaction at 160-200℃ for 12-24 hours, cooling, washing and drying, dispersing the product in deionized water, adding ferric chloride and PVP and stirring, adding concentrated ammonia at 70-80℃, continuing to stir for 1-2 hours, cooling, centrifuging to separate the microspheres, washing and drying to obtain magnetic chitosan microspheres, and encapsulating the probiotics in the magnetic chitosan microspheres by a swelling-adsorption method;

[0027] The solid-liquid ratio of chitosan to water is 1:10-20 g / mL, and the solid-liquid ratio of the product to deionized water is 1:10-20 g / mL; the mass ratio of ferric chloride, PVP, and the product is 1-2:0.5-0.7:5; the amount of concentrated ammonia added is 0.2-0.4 times the mass of the product, and the concentration is 20-30 wt%; the loading of probiotics on the magnetic chitosan microspheres is 20%-30%.

[0028] Furthermore, in step S6, during the initial cultivation, the salinity of the culture pond is controlled at 15-25‰, the temperature at 25-28℃, the pH value at 7.5-8.0, and the dissolved oxygen at greater than 6.0 mg / L.

[0029] Compared with the prior art, the beneficial effects of the present invention are:

[0030] 1. In the female parent fish rearing and maturation stage of the present invention, by controlling the feeding of specific biological feed in stages, it can not only promote the feeding of parent fish and accelerate growth, but also, in conjunction with the specific formulated feed in the pre-spawning control stage, create favorable conditions for subsequent spawning and improve the quality and quantity of eggs.

[0031] 2. This invention introduces low-frequency ultrasound to assist fertilization during the artificial insemination stage, which can significantly improve the vitality of eggs and sperm and increase the fertilization rate; during the hatching stage of fertilized eggs, intermittent ultrasound can be used to improve the hatching rate and increase the yield of golden tiger hybrid seedlings.

[0032] 3. After hatching, the fish fry of this invention are transferred to a rearing pond containing probiotic magnetic chitosan microspheres. The probiotics are released slowly, reducing loss and improving the adaptability of the fish fry. At the same time, the probiotics can adsorb harmful substances, reduce pathogens, stabilize water quality, promote the growth of fish fry, improve the survival rate, reduce the deformity rate, and improve the quality of golden tiger hybrid fry. In addition, the magnetic field can be used to control the release location of probiotics, so that the probiotics can be evenly distributed. Detailed Implementation

[0033] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0034] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0035] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0036] The preparation of biological feed B of the present invention is to mix the various raw materials evenly, puff and granulate, grind and sieve; the preparation of biological feed A is to mix the various raw materials evenly, puff and granulate, grind and sieve; the preparation of compound feed is to mix the various raw materials evenly, puff and granulate, grind and sieve.

[0037] The extracts of the present invention are: * **Cephalotaxus fortunei** (a type of fruit) aqueous extract and / or * **Cephalotaxus fortunei** ethanol extract; * **Dendrobium nobile** (a type of herb) stem extract is * **Dendrobium nobile** stem ethanol extract and / or * **Dendrobium nobile** stem ethyl acetate extract; * **Dioscorea opposita** (a type of fruit) fruit extract is * **Dioscorea opposita** fruit aqueous extract and / or * **Dioscorea opposita** fruit ethanol extract; * **Dendrobium nobile** (a type of herb) extract is * **Dendrobium nobile** herb aqueous extract and / or * **Dendrobium nobile** herb ethanol extract; * **Ganoderma lucidum** (a type of mushroom) extract is * **Ganoderma lucidum** herb aqueous extract and / or * **Ganoderma lucidum** herb ethanol extract; * **Perilla frutescens** (a type of herb) leaf extract is * **Perilla frutescens** leaf aqueous extract and / or * **Perilla frutescens** leaf ethanol extract; * **Morinda officinalis** (a type of fruit) fruit extract is * **Morinda officinalis** fruit ethanol extract and / or * **Morinda officinalis** fruit ethyl acetate extract.

[0038] The earthworm fermentation liquid of the present invention is obtained by pulverizing earthworms and then ultrasonically stirring with lipase; the chitin fermentation liquid is obtained by crushing shrimp shells and then fermenting them with Trichoderma harzianum and Bacillus spp. The preferred mass ratio of Trichoderma harzianum to Bacillus spp. is 1:1, and the preferred fermentation temperature is 35-45℃.

[0039] Example 1

[0040] The seedling cultivation method for Golden Tiger Hybrid Spotted Varieties includes the following steps:

[0041] S1. Optimize the breeding environment: Select a pond with a water temperature of 23℃ and a water depth of 2 meters, and stock it with healthy, disease-free brown-spotted grouper that are over 6 years old, at a density of 150 fish / m³. 2 ;

[0042] S2. Female parent breeding and maturation: In the first stage, feed biological feed A at a rate of 5% of the female parent's body weight, 3 times a day for 5 days. In the second stage, feed a mixture of biological feed B and nutritional preparations at a rate of 6% of the female parent's body weight, 3 times a day for 4 days.

[0043] Biological feed A, by weight, includes the following ingredients: 35 parts fish meal, 8 parts rotifers, 2 parts vitamins, 2 parts amino acids, 5 parts green fruit extract, 2 parts nightshade polysaccharide, 5 parts Dendrobium nobile stem extract, and 3 parts coconut oil.

[0044] Biological feed B, by weight, includes the following ingredients: 25 parts fish meal, 5 parts shrimp meal, 2 parts vitamins, 2 parts tea oil polysaccharide, 2 parts shiitake mushroom polysaccharide, 1.5 parts Dioscorea opposita fruit extract, 4 parts bamboo orchid extract, and 3 parts blended vegetable oil.

[0045] The nutrient preparation comprises the following raw materials in parts by weight: 4 parts earthworm fermentation liquid and 4 parts chitin fermentation liquid; the nutrient preparation is prepared by treating the earthworm fermentation liquid and chitin fermentation liquid twice at 100 MPa using a dynamic high-pressure micro-jet homogenizer and then mixing them.

[0046] S3. Pre-spawning management: Six days before the female parent lays eggs, raise the water temperature to 27°C and feed them with formulated feed at a rate of 2% of the female parent's body weight, twice a day.

[0047] The compound feed, by weight, includes the following ingredients: 15 parts shrimp powder, 7 parts clam meat paste, 3 parts oxalic acid nutrient solution, 3.5 parts wild Ganoderma lucidum extract, 2 parts perilla leaf extract, 2 parts Morinda officinalis fruit extract, and 2 parts longan polysaccharide.

[0048] S4. Fertilization: After the female parent lays eggs, the eggs are fertilized with sperm from the blue-bodied large-spotted grouper under low-frequency ultrasonic field conditions to obtain fertilized eggs.

[0049] The specific conditions for the low-frequency ultrasound field are as follows: ultrasound at 25kHz and 13W for 25 seconds, pause for 30 seconds, and then continue ultrasound for 25 seconds. Repeat the above process until fertilization is completed.

[0050] S5. Hatching of fertilized eggs: Place the fertilized eggs in an incubator and sonicate them at 25 kHz and 25 W for 40 seconds with a 3-minute interval. Then sonicate them at 55 kHz and 55 W for 30 seconds with a 5-minute interval. Repeat the above process 4 times to hatch the fry.

[0051] S6. Fry rearing: Select a rearing pond with a salinity of 20‰, a temperature of 27℃, a pH of 7.7±0.2, and dissolved oxygen greater than 6.0mg / L. Add probiotic magnetic chitosan microspheres and transfer the fry to the rearing pond for initial rearing.

[0052] The preparation of probiotic magnetic chitosan microspheres involved the following steps: Chitosan was added to water at a solid-liquid ratio of 1:15 g / mL, then transferred to a high-pressure reactor and subjected to a hydrothermal reaction at 180℃ for 12 hours. After cooling, the microspheres were washed and dried. The product was then dispersed in deionized water at a solid-liquid ratio of 1:15 g / mL. Ferric chloride and PVP were added, with a mass ratio of ferric chloride, PVP, and product of 1:0.6:5. The mixture was stirred, and 25 wt% concentrated ammonia was added at 75℃. The amount of concentrated ammonia added was 0.3 times the mass of the product. The mixture was stirred for another 1.5 hours, cooled, and the microspheres were separated by centrifugation, washed, and dried to obtain magnetic chitosan microspheres. Probiotics were then encapsulated in the magnetic chitosan microspheres using a swelling-adsorption method, with a probiotic loading of 25% on the magnetic chitosan microspheres.

[0053] Example 2

[0054] The seedling cultivation method for Golden Tiger Hybrid Spotted Varieties includes the following steps:

[0055] S1. Optimize the breeding environment: Select a pond with a water temperature of 22℃ and a water depth of 2 meters, and stock it with healthy, disease-free brown-spotted grouper that are over 6 years old, at a density of 100 fish / m³. 2 ;

[0056] S2. Female parent breeding and maturation: In the first stage, feed biological feed A at a rate of 4% of the female parent's body weight, 3 times a day for 4 days. In the second stage, feed a mixture of biological feed B and nutritional preparations at a rate of 5% of the female parent's body weight, 2 times a day for 3 days.

[0057] Biological feed A, by weight, includes the following ingredients: 30 parts fish meal, 5 parts rotifers, 1 part vitamins, 1 part amino acids, 4 parts green fruit extract, 1 part nightshade polysaccharide, 4 parts Dendrobium nobile stem extract, and 2 parts coconut oil.

[0058] Biological feed B, by weight, includes the following ingredients: 20 parts fish meal, 4 parts shrimp meal, 1 part vitamin, 1 part tea oil polysaccharide, 1 part shiitake mushroom polysaccharide, 1 part Dioscorea opposita fruit extract, 3 parts bamboo orchid extract, and 2 parts blended vegetable oil.

[0059] The nutrient preparation comprises the following raw materials in parts by weight: 3 parts earthworm fermentation liquid and 3 parts chitin fermentation liquid; the nutrient preparation is prepared by treating the earthworm fermentation liquid and chitin fermentation liquid twice at 80 MPa using a dynamic high-pressure micro-jet homogenizer and then mixing them.

[0060] S3. Pre-spawning management: Five days before the female parent lays eggs, raise the water temperature to 27°C and feed them with formulated feed at a rate of 1% of the female parent's body weight, twice a day.

[0061] The compound feed, by weight, includes the following ingredients: 10 parts shrimp powder, 5 parts clam meat paste, 2 parts sarsaparilla acid nutrient solution, 3 parts wild Ganoderma lucidum extract, 1 part perilla leaf extract, 1 part Morinda officinalis fruit extract, and 1 part longan polysaccharide.

[0062] S4. Fertilization: After the female parent lays eggs, the eggs are fertilized with sperm from the blue-bodied large-spotted grouper under low-frequency ultrasonic field conditions to obtain fertilized eggs.

[0063] The specific conditions for the low-frequency ultrasound field are as follows: ultrasound at 20kHz and 10W for 20 seconds, pause for 30 seconds, and then continue ultrasound for 20 seconds. Repeat the above process until fertilization is completed.

[0064] S5. Hatching of fertilized eggs: Place the fertilized eggs in an incubator and sonicate them at 20kHz and 20W for 30 seconds with a 3-minute interval. Then sonicate them at 50kHz and 50W for 20 seconds with a 5-minute interval. Repeat the above process 3 times to hatch the fry.

[0065] S6. Fry rearing: Select a rearing pond with a salinity of 15‰, a temperature of 25℃, a pH of 7.7±0.2, and dissolved oxygen greater than 6.0mg / L. Add probiotic magnetic chitosan microspheres and transfer the fry to the rearing pond for initial rearing.

[0066] The preparation of probiotic magnetic chitosan microspheres involved the following steps: Chitosan was added to water at a solid-liquid ratio of 1:10 g / mL, then transferred to a high-pressure reactor and subjected to a hydrothermal reaction at 160℃ for 12 hours. After cooling, the microspheres were washed and dried. The product was then dispersed in deionized water at a solid-liquid ratio of 1:10 g / mL, and ferric chloride and PVP were added. The mass ratio of ferric chloride, PVP, and product was 1:0.5:5. The mixture was stirred, and 20 wt% concentrated ammonia was added at 70℃. The amount of concentrated ammonia added was 0.2 times the mass of the product. The mixture was stirred for another hour, cooled, and the microspheres were separated by centrifugation, washed, and dried to obtain magnetic chitosan microspheres. Probiotics were then encapsulated in the magnetic chitosan microspheres using a swelling-adsorption method, with a probiotic loading of 20% on the magnetic chitosan microspheres.

[0067] Example 3

[0068] The seedling cultivation method for Golden Tiger Hybrid Spotted Varieties includes the following steps:

[0069] S1. Selecting a breeding environment: Choose a pond with a water temperature of 25℃ and a water depth of 3 meters. Stock the pond with healthy, disease-free brown-spotted grouper that are over 6 years old at a density of 200 fish / m³. 2 ;

[0070] S2. Female parent breeding and maturation: In the first stage, feed biological feed A at a rate of 6% of the female parent's body weight, 4 times a day for 6 days. In the second stage, feed a mixture of biological feed B and nutritional preparations at a rate of 7% of the female parent's body weight, 3 times a day for 5 days.

[0071] Biological feed A, by weight, includes the following ingredients: 40 parts fish meal, 10 parts rotifers, 3 parts vitamins, 3 parts amino acids, 6 parts green fruit extract, 3 parts nightshade polysaccharide, 6 parts Dendrobium nobile stem extract, and 4 parts coconut oil.

[0072] Biological feed B, by weight, includes the following ingredients: 30 parts fish meal, 6 parts shrimp meal, 3 parts vitamins, 3 parts tea oil polysaccharide, 3 parts shiitake mushroom polysaccharide, 2 parts Dioscorea bulbifera fruit extract, 5 parts bamboo orchid extract, and 4 parts blended vegetable oil.

[0073] The nutrient preparation, by weight, includes the following raw materials: 5 parts earthworm fermentation liquid and 5 parts chitin fermentation liquid; the nutrient preparation is prepared by treating the earthworm fermentation liquid and chitin fermentation liquid twice at 120MPa using a dynamic high-pressure micro-jet homogenizer and then mixing them.

[0074] S3. Pre-spawning management: Seven days before the female parent lays eggs, raise the water temperature to 28°C and feed them with formulated feed at a rate of 3% of the female parent's body weight, twice a day.

[0075] The compound feed, by weight, includes the following ingredients: 20 parts shrimp powder, 8 parts clam meat paste, 4 parts sarsaparilla acid nutrient solution, 4 parts wild Ganoderma lucidum extract, 3 parts perilla leaf extract, 3 parts Morinda officinalis fruit extract, and 3 parts longan polysaccharide.

[0076] S4. Fertilization: After the female parent lays eggs, the eggs are fertilized with sperm from the blue-bodied large-spotted grouper under low-frequency ultrasonic field conditions to obtain fertilized eggs.

[0077] The specific conditions for the low-frequency ultrasound field are as follows: ultrasound at 30kHz and 15W for 30 seconds, pause for 30 seconds, and then continue ultrasound for 30 seconds. Repeat the above process until fertilization is completed.

[0078] S5. Hatching of fertilized eggs: Place the fertilized eggs in an incubator and sonicate them at 30 kHz and 30 W for 60 seconds, with a 3-minute interval. Then sonicate them at 60 kHz and 60 W for 40 seconds, with a 5-minute interval. Repeat the above process 5 times to hatch the fish fry.

[0079] S6. Fry rearing: Select a rearing pond with a salinity of 25‰, a temperature of 28℃, a pH of 7.7±0.2, and dissolved oxygen greater than 6.0mg / L. Add probiotic magnetic chitosan microspheres and transfer the fry to the rearing pond for initial rearing.

[0080] The preparation of probiotic magnetic chitosan microspheres involved the following steps: Chitosan was added to water at a solid-liquid ratio of 1:20 g / mL, then transferred to a high-pressure reactor and subjected to a hydrothermal reaction at 200℃ for 24 hours. After cooling, the microspheres were washed and dried. The product was then dispersed in deionized water at a solid-liquid ratio of 1:20 g / mL, and ferric chloride and PVP were added. The mass ratio of ferric chloride, PVP, and product was 2:0.7:5. The mixture was stirred, and 30 wt% concentrated ammonia was added at 80℃. The amount of concentrated ammonia added was 0.4 times the mass of the product. The mixture was stirred for another 2 hours, cooled, and the microspheres were separated by centrifugation, washed, and dried to obtain magnetic chitosan microspheres. Probiotics were then encapsulated in the magnetic chitosan microspheres using a swelling-adsorption method, with a probiotic loading of 30% on the magnetic chitosan microspheres.

[0081] Comparative Example 1

[0082] The difference from Example 1 is that the biological feed A in step S2 lacks green fruit extract and nightshade polysaccharide, while the rest is the same as in Example 1.

[0083] Comparative Example 2

[0084] The difference from Example 1 is that in the second stage of step S2, only biological bait B is fed, while the rest is the same as in Example 1.

[0085] Comparative Example 3

[0086] The difference from Example 1 is that in step S3, the pre-spawning control is performed by replacing the compound feed with biological feed B and nutritional preparations, while the rest is the same as in Example 1.

[0087] Comparative Example 4

[0088] The difference from Example 1 is that the low-frequency ultrasonic field is missing in step S4, but otherwise it is the same as Example 1.

[0089] Comparative Example 5

[0090] The difference from Example 1 is that intermittent ultrasound treatment is missing in step S5, but otherwise it is the same as Example 1.

[0091] Comparative Example 6

[0092] The difference from Example 1 is that in step S6, the probiotic magnetic chitosan microspheres are replaced with probiotics, while the rest is the same as in Example 1.

[0093] Test case

[0094] Golden tiger hybrid grouper was bred according to the breeding methods of Examples 1-3 and Comparative Examples 1-6. Experimental groups 1-9 were set up, corresponding to Examples 1-3 and Comparative Examples 1-6 respectively. Each group had 300 female parent brown-spotted grouper that were 6 years or older, and the golden tiger hybrid grouper fry were obtained according to the corresponding breeding methods.

[0095] 1. Measure the egg production, fertilization rate, hatching rate and fry deformity rate respectively.

[0096] The results are shown in Table 1.

[0097] Table 1

[0098] Example 1 143.4Kg 99.5％ 97.5％ 0％ Example 2 138.6Kg 99.3％ 97.2％ 0％ Example 3 135.7Kg 99.1％ 96.9％ 0％ Comparative Example 1 112.5Kg 92.8％ 92.2％ 0.8％ Comparative Example 2 98.4Kg 91.1％ 91.5％ 1.1％ Comparative Example 3 105.6Kg 90.3％ 88.9％ 1.4％ Comparative Example 4 - 80.2％ 87.1％ 1.5％ Comparative Example 5 - - 80.6％ 2.4％ Comparative Example 6 - - - -

[0099] As shown in Table 1, the cultivation methods of Examples 1-3 of this invention, by feeding specific feed, can significantly increase the number and quality of eggs, and significantly improve the fertilization rate and hatching rate. No deformities were observed in the hatched fry. Comparative Examples 1-3, by changing the composition of the feed formula, resulted in a decrease in the number of eggs, a reduction in egg quality, and negatively impacted the subsequent fertilization rate, hatching rate, and deformity rate.

[0100] 2. After 5 months of breeding, the survival rate and disease rate were counted, and the results are shown in Table 2.

[0101] Table 2

[0102]

[0103]

[0104] As can be seen from Table 2, the artificial breeding method of the present invention can significantly improve the survival rate of fish fry, improve the quality of healthy fry, and the fish fry raised for 5 months are almost disease-free.

[0105] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for cultivating seedlings of the Golden Tiger Hybrid, characterized in that, Includes the following steps: S1. Select a breeding environment: Select a pond with a water temperature of 22-25℃ and a water depth of 2-3 meters, and release female parent brown-spotted grouper. S2. Female parent breeding and maturation: In the first stage, feed biological feed A at a rate of 4%-6% of the female parent's body weight, 3-4 times a day for 4-6 days. In the second stage, feed a mixture of biological feed B and nutritional preparations at a rate of 5%-7% of the female parent's body weight, 2-3 times a day for 3-5 days. The first stage of feeding is live feed A, which includes the following ingredients by weight: 30-40 parts fish meal, 5-10 parts rotifers, 1-3 parts vitamins, 1-3 parts amino acids, 4-6 parts green fruit extract, 1-3 parts nightshade polysaccharide, 4-6 parts Dendrobium nobile stem extract, and 2-4 parts coconut oil. The second stage involves feeding with biological feed B, which, by weight, includes the following ingredients: 20-30 parts fish meal, 4-6 parts shrimp meal, 1-3 parts vitamins, 1-3 parts tea oil polysaccharide, 1-3 parts shiitake mushroom polysaccharide, 1-2 parts Dioscorea bulbifera fruit extract, 3-5 parts bamboo orchid extract, and 2-4 parts blended vegetable oil. The nutritional preparation comprises, by weight, the following raw materials: 3-5 parts earthworm fermentation liquid and 3-5 parts chitin fermentation liquid; S3. Pre-spawning management: 5-7 days before the female parent lays eggs, raise the water temperature to 27-28℃ and feed them with formulated feed at a rate of 1%-3% of the female parent's body weight, twice a day. The compound feed, by weight, includes the following ingredients: 10-20 parts shrimp powder, 5-8 parts clam meat paste, 2-4 parts oxalic acid nutrient solution, 3-4 parts wild Ganoderma lucidum extract, 1-3 parts perilla leaf extract, 1-3 parts Morinda officinalis fruit extract, and 1-3 parts longan polysaccharide. S4. Fertilization: After the female parent lays eggs, the eggs are fertilized with sperm from the blue-bodied large-spotted grouper under low-frequency ultrasonic field conditions to obtain fertilized eggs. S5. Fertilized egg hatching: Fertilized eggs are placed in an incubator and subjected to intermittent ultrasonic treatment to hatch into fish fry; S6. Fish fry rearing: Select a rearing pond, add probiotic magnetic chitosan microspheres, and transfer the fish fry to the rearing pond for initial rearing.

2. The method for cultivating seedlings of the Golden Tiger hybrid as described in claim 1, characterized in that, In step S1, the female parent brown-spotted grouper are selected from healthy brown-spotted groupers that are over 6 years old and free from disease, and the stocking density is 100-200 fish / m². 2 .

3. The method for cultivating seedlings of the Golden Tiger hybrid as described in claim 1, characterized in that, In step S2, the preparation method of the nutrient preparation is as follows: earthworm fermentation liquid and chitin fermentation liquid are treated twice by a dynamic high-pressure micro-jet homogenizer at 80-120MPa, and then mixed to obtain the nutrient preparation.

4. The method for cultivating seedlings of the Golden Tiger hybrid as described in claim 1, characterized in that, In step S4, the low-frequency ultrasound field treatment specifically involves: performing ultrasound at 20-30 kHz and 10-15 W for 20-30 seconds, followed by a 30-second interval, and then continuing ultrasound for another 20-30 seconds. This process is repeated until fertilization is complete.

5. The method for cultivating seedlings of the Golden Tiger Hybrid as described in claim 1, characterized in that, In step S5, the intermittent ultrasound treatment is to first use ultrasound at 20-30kHz and 20-30W for 30-60s, with a 3-min interval, then use ultrasound at 50-60kHz and 50-60W for 20-40s, with a 5-min interval, and repeat the above process 3-5 times.

6. The method for cultivating seedlings of the Golden Tiger hybrid as described in claim 1, characterized in that, In step S6, the preparation method of the probiotic magnetic chitosan microspheres includes: adding chitosan to water, transferring it to a high-pressure reactor, carrying out a hydrothermal reaction at 160-200℃ for 12-24 hours, cooling, washing and drying, dispersing the product in deionized water, adding ferric chloride and PVP and stirring, adding concentrated ammonia at 70-80℃, continuing to stir for 1-2 hours, cooling, centrifuging to separate the microspheres, washing and drying to obtain magnetic chitosan microspheres, and encapsulating the probiotics in the magnetic chitosan microspheres by a swelling-adsorption method; The solid-liquid ratio of chitosan to water is 1:10-20 g / mL, and the solid-liquid ratio of the product to deionized water is 1:10-20 g / mL; the mass ratio of ferric chloride, PVP to the product is 1-2:0.5-0.7:5; the amount of concentrated ammonia added is 0.2-0.4 times the mass of the product, and the concentration is 20-30 wt%; the loading of probiotics on the magnetic chitosan microspheres is 20%-30%.

7. The method for cultivating seedlings of the Golden Tiger hybrid as described in claim 1, characterized in that, In step S6, during the initial cultivation, the salinity of the culture pond is controlled at 15-25‰, the temperature at 25-28℃, the pH at 7.5-8.0, and the dissolved oxygen at 6.0mg / L.

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