Cultivation method of golden and tiger hybrid spotted fish eggs

Abstract

The invention provides a breeding method of golden and tiger hybrid spotted fish eggs. The breeding method comprises the following steps: S1, parent screening; s2, parent cultivation; s3, artificial insemination: (1) spawning induction: when the belly of the female parent fish expands, the ovary develops to the IV stage and the male parent fish presses the belly to cause white seminal fluid to flow out, performing oxygenation flowing water stimulation on the female parent fish, and performing irradiation with a TDP radiation lamp for spawning induction; (2) fertilization; s4, artificial incubation: (1) arranging an incubator, injecting seawater, and performing plasma treatment to obtain a treated incubator; and (2) adding the fertilized eggs into the treated incubator, carrying out magnetization treatment, adding chitosan, and hatching to obtain the larva fish. According to the breeding method, the sperm and egg quality can be improved, the fertilization rate and the hatching rate are remarkably increased, and according to verification of the embodiment, the fertilization rate ranges from 97.4% to 99.3%, and the hatching rate ranges from 95.6% to 97.1%. The hatched larvae are not deformed, the survival rate is 96%-99%, the quality of the larvae is high, and a good foundation is established for obtaining high-quality hybrid Jinhuasu.

Description

Technical Field

[0001] The invention relates to the technical field of aquatic product cultivation, and in particular to a method for cultivating golden tiger hybrid grouper eggs. Background Art

[0002] Brown-spotted grouper (scientific name: Epinephelus fuscoguttatus ), belonging to the order Perciformes, family Serranidae, genus Epinephelus, also known as tiger grouper, brown-spotted grouper, inhabits a depth of 1-60 meters, can reach a body length of 120 cm, is a warm-water nearshore and coral reef fish, and lives in coral reefs and lagoons with clear water. It is carnivorous, feeding on fish, cephalopods, and crustaceans, and can be used as edible fish, farmed fish, and ornamental fish. The brown-spotted grouper is a fierce feeder and grows fast, and is widely used in aquaculture production.

[0003] Blue-bodied grouper (scientific name: Epinephelus tukula ), commonly known as the golden grouper and black-spotted grouper, is a fish of the genus Epinephelus in the order Perciformes, suborder Perciformes, family Serranidae, and is second only to the saddle-banded grouper in size. It grows fast and is a grouper species with great breeding prospects. It is distributed in the Indo-West Pacific region, from the Red Sea, East Africa to the waters of Queensland, Australia, and inhabits depths of 10-150 meters. It can reach a body length of 200 cm and a weight of 110 kg. It is a world-famous warm-water fish.

[0004] Golden Tiger Hybrid Grouper is a grouper obtained by hybridizing brown spotted grouper as the mother and saddle band grouper and blue body big spotted grouper as the father. However, the conventional breeding methods currently used will have the following problems: low fertilization rate and hatching rate, some fry after hatching are deformed, and the survival rate of fry is low, which affects the yield and quality of Golden Tiger Hybrid Grouper fry, and seriously restricts the healthy development of the grouper industry. Summary of the invention

[0005] In view of this, the present invention provides a method for cultivating golden tiger hybrid grouper eggs.

[0006] The technical solution of the present invention is achieved in this way: A method for cultivating golden tiger hybrid grouper eggs comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free, high-metabolism grouper as the male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond area of ​​2-3 mu, a water depth of 2-3m, a stocking density of 50-150 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; (2) Male parent breeding: Set up a pond area of ​​4-5 mu, a water depth of 4-5m, a stocking density of 300-400 fish / mu, and carry out water quality acclimation and low temperature acclimation in sequence; S3. Artificial insemination (1) Inducing spawning: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated flowing water and irradiated with TDP radiation lamps to induce spawning; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) setting up an incubator, injecting seawater, and performing plasma treatment to obtain a treated incubator; (2) The fertilized eggs are placed in a treated incubator and subjected to magnetization treatment. During the magnetization treatment, water-soluble chitosan is added to obtain fry through hatching.

[0007] Furthermore, in steps S2 (1) and (2), the pond is provided with an inlet and an outlet, has an adequate water source, is pollution-free, and is aerated every 5-7 days.

[0008] Furthermore, in step S2 (1), the method for acclimating the water quality includes: first setting the pond water temperature to 22-30°C, the dissolved oxygen to 5-7 mg / L, the light to 700-900 Lux, and 5-7 days before the culture, setting the salinity to 15-20‰, the water flow rate to 0.3-0.6 m 3 / h, for 10-12 hours a day, then adjust the salinity to 22-28‰ and the water flow rate to 1.2-1.5m 3 / h, lasting 6-8h per day, and acclimation for 2-4 days; The nutritional domestication method comprises: feeding the female broodstock with water quality domestication bait a in an amount of 3%-5% of the female broodstock's body weight, 4-5 times a day, and continuously feeding for 4-6 days, and then feeding bait b in an amount of 6%-8% of the female broodstock's body weight, 2-3 times a day for domestication for 4-6 days.

[0009] Furthermore, in the nutritional domestication, the bait a comprises the following raw materials in parts by weight: 8-10 parts of fish meal, 2-4 parts of dried shrimp, 1-3 parts of oysters, 1-2 parts of corn flour, 0.8-1.2 parts of soybean lecithin, 1-3 parts of Artemia nauplii and 0.2-0.4 parts of vitamins; The bait B comprises the following raw materials in parts by weight: 15-20 parts of fish meal, 0.5-0.7 parts of soy isoflavones, 5-7 parts of peanut meal, 1-3 parts of pomegranate peel extract, 3-5 parts of acerola cherry extract, 2-4 parts of vegetable blending oil, 0.2-0.4 parts of astaxanthin, 0.6-0.8 parts of fucose and 0.2-0.4 parts of kudzu root polysaccharide.

[0010] Furthermore, in step S2 (2), the method for taming the water quality comprises: firstly setting the pond water temperature to 25-28°C, the dissolved oxygen to 6-10 mg / L, the light to 1000-1200 Lux, the salinity to 18-24‰, and the water flow rate to 0.6-0.8 m 3 / h, lasting 8-10h a day, after 5-7 days of culture, adjust the salinity to 26-30‰, and the water flow rate to 1.5-2.0m 3 / h, lasting 5-7h per day, and acclimation for 2-4 days; The low-temperature acclimation method comprises: adjusting the water temperature of the acclimated pond to 20-24° C., setting the water temperature to 15-19° C. after culturing for 3-5 days, and acclimating for 2-4 days.

[0011] Furthermore, in step S3 (1), the oxygenated water stimulation is a dissolved oxygen of 11-13 mg / L and a water flow rate of 2.2-2.4 m / s. 3 / h, maintain for 2-3 days.

[0012] Furthermore, in step S3 (1), the irradiation power of the TDP radiation lamp is 100-200W, and the irradiation time is 2-4h.

[0013] Furthermore, in step S4 (1), the plasma treatment is performed at 20-50 W and 20-30 kHz for 5-15 min.

[0014] Furthermore, in step S4 (2), the molecular weight of the water-soluble chitosan is 10-30 kDa, and the amount added is 0.1%-0.3%.

[0015] Furthermore, in step S4 (2), the magnetization treatment is performed at 100-200 Gs for 10-15 min.

[0016] Compared with the prior art, the present invention has the following beneficial effects: 1. The breeding method of the present invention improves the quality of sperm and eggs, and significantly improves the fertilization rate and hatching rate. The fertilization rate of the present invention is 97.4%-99.3%, and the hatching rate is 95.6%-97.1%. The fry after hatching does not have deformity, and the fry survival rate is 96%-99%. The seedling quality is high, and a good foundation is established for obtaining high-quality hybrid golden tiger grouper.

[0017] 2. The present invention improves reproductive capacity by strictly controlling the breeding environment and domestication methods in the cultivation and management of female and male parents, thus laying a good foundation for subsequent induction of labor and fertilization.

[0018] 3. In the inducing spawning process of the present invention, by adding TDP radiation lamp irradiation treatment to the female broodstock through oxygenated flowing water stimulation, the spawning efficiency and egg quality are improved, thereby improving the fertilization rate.

[0019] 4. Before the fertilized eggs are hatched, the incubator is subjected to plasma treatment to improve water quality, and subsequently the vitality of the fertilized eggs is improved, thereby improving the hatching rate; during the hatching process, a thin film is formed on the surface of the fertilized eggs through magnetization treatment and the addition of water-soluble chitosan, which can not only resist the adverse effects of the external environment, but also enhance the vitality of the fertilized eggs and improve the hatching rate, and at the same time provide nutritional support for the fry obtained after the fertilized eggs are hatched, promote the healthy growth of the fry, thereby improving the survival rate, and contributing to the economic benefits and ecological benefits of the aquaculture industry. DETAILED DESCRIPTION

[0020] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0021] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0022] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0023] The ponds of the present invention are all provided with inlet and outlet drainage ports, with sufficient water source and pollution-free water quality, and are aerated every 5-7 days.

[0024] The TDP radiation lamp of the present invention is a QG-22A type specific electromagnetic wave TDP radiation lamp.

[0025] The plasma treatment of the present invention is to connect the incubator and the low-temperature plasma generator through a pipeline, and introduce the plasma gas into the water to contact the water body.

[0026] The magnetization treatment of the present invention is achieved by a magnetized water treatment device, which is purchased from Shandong Fengshuo Environmental Protection Technology Co., Ltd.

[0027] Example 1 The breeding method of the golden tiger hybrid grouper roe of the present embodiment comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0028] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0029] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 150 Gs for 13 min. During this period, water-soluble chitosan with a molecular weight of 20 kDa was added. The amount of water-soluble chitosan added was 0.2% of the water in the incubator, and then the fry were hatched.

[0030] Example 2 The breeding method of the golden tiger hybrid grouper roe of the present embodiment comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 2 m, a stocking density of 50 per mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 22°C, dissolved oxygen to 5 mg / L, light to 700 Lux, and 5 days before breeding, setting the salinity to 15‰ and the water flow rate to 0.3 m 3 / h, for 10 hours a day, then adjust the salinity to 22‰ and the water flow rate to 1.2m 3 / h, for 6 hours a day, for 2 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 3% of the female broodstock's body weight, feeding 4 times a day, after feeding continuously for 4 days, feeding with bait b, the feeding amount is 6% of the female broodstock's body weight, feeding twice a day for 4 days of domestication.

[0031] Bait a comprises the following raw materials in parts by weight: 8 parts of fish meal, 2 parts of dried shrimps, 1 part of oyster, 1 part of corn flour, 0.8 parts of soybean lecithin, 1 part of Artemia nauplii and 0.2 parts of vitamins; bait b comprises the following raw materials in parts by weight: 15 parts of fish meal, 0.5 parts of soybean isoflavones, 5 parts of peanut meal, 1 part of pomegranate peel extract, 3 parts of acerola cherry extract, 2 parts of vegetable blending oil, 0.2 parts of astaxanthin, 0.6 parts of fucose and 0.2 parts of kudzu polysaccharide.

[0032] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 4 m, a stocking density of 300 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 25°C, dissolved oxygen to 6 mg / L, light to 1000 Lux, salinity to 18‰, water flow rate to 0.6 m3 / h, lasting 8 hours a day, after 5 days of culture, the salinity was adjusted to 26‰ and the water flow rate was 1.5m 3 / h, for 5 hours a day, for 2 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the acclimated pond to 20°C, setting the water temperature to 15°C after 3 days of cultivation, and acclimating for 2 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 11 mg / L, water flow rate 2.2 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 100W irradiation for 2h; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 20 W, 20 kHz for 5 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 100 Gs for 10 min. During this period, water-soluble chitosan with a molecular weight of 10 kDa was added. The amount of water-soluble chitosan added was 0.1% of the water in the incubator, and then the fry were hatched.

[0033] Example 3 The breeding method of the golden tiger hybrid grouper roe of the present embodiment comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​3 mu, a water depth of 3 m, and a stocking density of 150 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 30℃, dissolved oxygen to 7mg / L, light to 900Lux, and 7 days before breeding, setting the salinity to 20‰ and the water flow rate to 0.6m 3 / h, for 12 hours a day, then adjust the salinity to 28‰ and the water flow rate to 1.5m 3 / h, lasting 8h per day, for 4 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 5% of the female broodstock's body weight, feeding 5 times a day, and continuing feeding for 6 days, and then feeding with bait b, the feeding amount is 8% of the female broodstock's body weight, feeding 3 times a day for 6 days.

[0034] Bait a comprises the following raw materials in parts by weight: 10 parts of fish meal, 4 parts of dried shrimps, 3 parts of oysters, 2 parts of corn flour, 1.2 parts of soybean lecithin, 3 parts of Artemia nauplii and 0.4 parts of vitamins; bait b comprises the following raw materials in parts by weight: 20 parts of fish meal, 0.7 parts of soybean isoflavones, 7 parts of peanut meal, 3 parts of pomegranate peel extract, 5 parts of acerola cherry extract, 4 parts of vegetable blending oil, 0.4 parts of astaxanthin, 0.8 parts of fucose and 0.4 parts of kudzu polysaccharide.

[0035] (2) Male parent breeding: Set up a pond with an area of ​​5 mu, a water depth of 5 m, a stocking density of 400 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality acclimation includes: first setting the pond water temperature to 28°C, dissolved oxygen to 10 mg / L, light to 1200 Lux, salinity to 24‰, water flow rate to 0.8 m 3 / h, lasting 10 hours a day, after 7 days of culture, adjust the salinity to 30‰ and the water flow rate to 2.0m 3 / h, for 7h a day, for 4 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the acclimated pond to 24°C, setting the water temperature to 19°C after 5 days of cultivation, and acclimating for 4 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 13 mg / L, water flow rate 2.4 m / s 3 / h, maintained for 3 days, during which time TDP radiation lamps were added above the pond to induce spawning, specifically: 200W irradiation for 4 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 50 W, 30 kHz for 15 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 200 Gs for 15 min. During this period, water-soluble chitosan with a molecular weight of 30 kDa was added. The amount of water-soluble chitosan added was 0.3% of the water in the incubator. The fry were then hatched to obtain fry.

[0036] Comparative Example 1 The difference from Example 1 is that the female parent cultivation lacks nutritional domestication, and the rest is consistent with Example 1.

[0037] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond area of ​​2 mu, a water depth of 3m, a stocking density of 100 fish / mu, and carry out water quality acclimatization. Specifically, first set the pond water temperature to 28℃, dissolved oxygen to 6mg / L, and light to 800Lux. Six days before breeding, set the salinity to 18‰ and the water flow rate to 0.4m 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 150 Gs for 13 min. During this period, water-soluble chitosan with a molecular weight of 20 kDa was added. The amount of water-soluble chitosan added was 0.2% of the water in the incubator, and then the fry were hatched.

[0038] Comparative Example 2 The difference from Example 1 is that the male parent cultivation lacks low temperature acclimation, and the rest is consistent with Example 1.

[0039] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0040] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0041] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 150 Gs for 13 min. During this period, water-soluble chitosan with a molecular weight of 20 kDa was added. The amount of water-soluble chitosan added was 0.2% of the water in the incubator, and then the fry were hatched.

[0042] Comparative Example 3 The difference from Example 1 is that no TDP radiation lamp irradiation is performed during the artificial insemination and induction stage, and the rest is the same as Example 1.

[0043] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0044] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0045] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintain for 2 days; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs were added to the treated incubator and magnetized at 150 Gs for 13 min. During this period, water-soluble chitosan with a molecular weight of 20 kDa was added. The amount of water-soluble chitosan added was 0.2% of the water in the incubator, and then the fry were hatched.

[0046] Comparative Example 4 The difference from Example 1 is that no plasma treatment is performed during the artificial incubation stage, and the rest is the same as Example 1.

[0047] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0048] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0049] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation An incubator was set up, seawater was injected, fertilized eggs were added into the incubator, and magnetized at 150 Gs for 13 minutes. During this period, water-soluble chitosan with a molecular weight of 20 kDa was added. The amount of water-soluble chitosan added was 0.2% of the water in the incubator, and then fry were hatched.

[0050] Comparative Example 5 The difference from Example 1 is that there is no magnetization in the artificial incubation stage, and the rest is the same as Example 1.

[0051] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0052] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0053] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs are added to the treated incubator, and water-soluble chitosan with a molecular weight of 20 kDa is added, the amount of water-soluble chitosan added is 0.2% of the water in the incubator, and then hatched to obtain fry.

[0054] Comparative Example 6 The difference from Example 1 is that no water-soluble chitosan is added during the artificial incubation stage, and the rest is the same as Example 1.

[0055] The method for cultivating the golden tiger hybrid grouper eggs of this comparative example comprises the following steps: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond with an area of ​​2 mu, a water depth of 3 m, and a stocking density of 100 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; The method of water quality domestication includes: first setting the pond water temperature to 28°C, dissolved oxygen to 6 mg / L, light to 800 Lux, and setting the salinity to 18‰ and water flow rate to 0.4 m / s for 6 days before breeding. 3 / h, for 11 hours a day, then adjust the salinity to 25‰ and the water flow rate to 1.4m 3 / h, for 7h a day, for 3 days of acclimatization; The method of nutritional domestication includes: feeding the female broodstock that have been domesticated with water quality with bait a, the feeding amount is 4% of the female broodstock's body weight, feeding 5 times a day, after feeding continuously for 5 days, feeding with bait b, the feeding amount is 7% of the female broodstock's body weight, feeding 3 times a day for 5 days of domestication.

[0056] Bait a comprises the following raw materials in parts by weight: 9 parts of fish meal, 3 parts of dried shrimps, 2 parts of oysters, 1.5 parts of corn flour, 1 part of soybean lecithin, 2 parts of Artemia nauplii and 0.3 parts of vitamins; bait b comprises the following raw materials in parts by weight: 18 parts of fish meal, 0.6 parts of soybean isoflavones, 6 parts of peanut meal, 2 parts of pomegranate peel extract, 4 parts of acerola cherry extract, 3 parts of vegetable blending oil, 0.3 parts of astaxanthin, 0.7 parts of fucose and 0.3 parts of kudzu polysaccharide.

[0057] (2) Male parent breeding: Set up a pond with an area of ​​4 mu, a water depth of 5 m, a stocking density of 350 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in sequence; The method of water quality domestication includes: first setting the pond water temperature to 27°C, dissolved oxygen to 8 mg / L, light to 1100 Lux, salinity to 20‰, water flow rate to 0.7 m 3 / h, for 9 hours a day, after 6 days of culture, the salinity was adjusted to 28‰ and the water flow rate was 1.8m 3 / h, for 6 hours a day, for 3 days of acclimatization; The method of low temperature acclimation includes: adjusting the water temperature of the pond after water acclimation to 22°C, setting the water temperature to 17°C after 4 days of cultivation, and acclimating for 3 days; S3. Artificial insemination (1) Induced labor: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated water flow, specifically: dissolved oxygen 12 mg / L, water flow rate 2.3 m / s 3 / h, maintained for 2 days, during which time a TDP radiation lamp was added above the pond to induce spawning, specifically: 150W irradiation for 3 hours; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) Setting up an incubator, injecting seawater, and performing plasma treatment, specifically: treating at 40 W, 25 kHz for 10 min to obtain a treated incubator; (2) The fertilized eggs were placed in a treated incubator, magnetized at 150 Gs for 13 min, and then hatched to obtain fry.

[0058] Test example According to the breeding methods of Examples 1-3 and Comparative Examples 1-6, golden tiger hybrid groupers were bred, and 9 groups were set up. Each group consisted of 250 female disease-free, metabolically-rich brown-spotted groupers (5-8 years old), and the egg-laying amount, fertilization rate, hatching rate and fry deformity rate were measured respectively. After 3 months of breeding, the fry survival rate was counted.

[0059] The results are shown in Table 1.

[0060] Table 1 As can be seen from Table 1, the breeding methods of Examples 1-3 of the present invention can increase the egg-laying amount, significantly improve the fertilization rate and hatching rate, obtain the golden tiger hybrid spotted fish fry without deformity, and improve the yield and quality of healthy seedlings.

[0061] 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 in the protection scope of the present invention.

Claims

1. A method for cultivating golden tiger hybrid grouper eggs, characterized in that: The following steps are involved: S1. Parent screening (1) Female parents: Select 5-8-year-old disease-free brown-spotted grouper with high metabolic rate as female parents; (2) Male parent: Select 3-5-year-old disease-free blue-bodied grouper with high metabolic rate as male parent; S2. Parent cultivation (1) Female parent breeding: Set up a pond area of ​​2-3 mu, a water depth of 2-3m, and a stocking density of 50-150 fish / mu, and carry out water quality domestication and nutrition domestication in sequence; (2) Male parent breeding: Set up a pond area of ​​4-5 mu, a water depth of 4-5m, a stocking density of 300-400 fish / mu, and carry out water quality acclimatization and low temperature acclimatization in turn; S3. Artificial insemination (1) Inducing spawning: When the abdomen of the female broodstock swells, the ovaries develop to stage IV, and white semen flows out when the male broodstock presses on the abdomen, the female broodstock are stimulated by oxygenated flowing water and irradiated with TDP radiation lamps to induce spawning; (2) Fertilization: Collecting eggs from female broodstock and sperm from male broodstock for artificial insemination to obtain fertilized eggs; S4. Artificial incubation (1) setting up an incubator, injecting seawater, and performing plasma treatment to obtain a treated incubator; (2) The fertilized eggs are placed in a treated incubator and subjected to magnetization treatment. During the magnetization treatment, water-soluble chitosan is added to obtain fry through hatching.

2. The method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, wherein: In steps S2 (1) and (2), the pond is provided with an inlet and outlet, has sufficient water source, has unpolluted water quality, and is aerated every 5-7 days.

3. The method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S2 (1), the method for acclimating the water quality includes: first setting the pond water temperature to 22-30°C, the dissolved oxygen to 5-7 mg / L, the light to 700-900 Lux, and 5-7 days before the culture, setting the salinity to 15-20‰, the water flow rate to 0.3-0.6 m 3 / h, for 10-12 hours a day, then adjust the salinity to 22-28‰ and the water flow rate to 1.2-1.5m 3 / h, lasting 6-8h per day, and acclimation for 2-4 days; The nutritional domestication method comprises: feeding the female broodstock with water quality domestication bait a in an amount of 3%-5% of the female broodstock's body weight, 4-5 times a day, and continuously feeding for 4-6 days, and then feeding bait b in an amount of 6%-8% of the female broodstock's body weight, 2-3 times a day for domestication for 4-6 days.

4. The method for cultivating a golden tiger hybrid grouper roe as claimed in claim 3, wherein: In the nutritional domestication, the bait a comprises the following raw materials in parts by weight: 8-10 parts of fish meal, 2-4 parts of dried shrimp, 1-3 parts of oysters, 1-2 parts of corn flour, 0.8-1.2 parts of soybean lecithin, 1-3 parts of Artemia nauplii and 0.2-0.4 parts of vitamins; The bait B comprises the following raw materials in parts by weight: 15-20 parts of fish meal, 0.5-0.7 parts of soy isoflavones, 5-7 parts of peanut meal, 1-3 parts of pomegranate peel extract, 3-5 parts of acerola cherry extract, 2-4 parts of vegetable blending oil, 0.2-0.4 parts of astaxanthin, 0.6-0.8 parts of fucose and 0.2-0.4 parts of kudzu root polysaccharide.

5. The method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S2 (2), the water quality acclimation method includes: first setting the pond water temperature to 25-28°C, the dissolved oxygen to 6-10 mg / L, the light to 1000-1200 Lux, the salinity to 18-24‰, and the water flow rate to 0.6-0.8 m 3 / h, lasting 8-10h a day, after 5-7 days of culture, adjust the salinity to 26-30‰, and the water flow rate to 1.5-2.0m 3 / h, lasting 5-7h per day, and acclimation for 2-4 days; The low-temperature acclimation method comprises: adjusting the water temperature of the acclimated pond to 20-24° C., setting the water temperature to 15-19° C. after culturing for 3-5 days, and acclimating for 2-4 days.

6. A method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S3 (1), the oxygenated water stimulation is a dissolved oxygen of 11-13 mg / L and a water flow rate of 2.2-2.4 m / s. 3 / h, maintain for 2-3 days.

7. A method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S3 (1), the irradiation power of the TDP radiation lamp is 100-200W, and the irradiation time is 2-4h.

8. A method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S4 (1), the plasma treatment is performed at 20-50 W and 20-30 kHz for 5-15 min.

9. A method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S4 (2), the molecular weight of the water-soluble chitosan is 10-30 kDa, and the amount added is 0.1%-0.3%.

10. The method for cultivating a golden tiger hybrid grouper egg as claimed in claim 1, characterized in that: In step S4 (2), the magnetization treatment is performed at 100-200 Gs for 10-15 min.

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