A method for artificial breeding of grouper fish by simulating ecology

Abstract

The present invention provides a kind of artificial breeding method of east star grouper fish by imitating ecology, and the steps are as follows: S1, select male and female broodstock to be placed in artificial imitating ecology cultivation pool for breeding; S2, in non-breeding season, feed female broodstock feed A2-3 times a day, feed male broodstock feed B 2-3 times a day; S3, 2-3 months before breeding, feed female broodstock fortified feed I 2-3 times a day, feed male broodstock fortified feed II 2-3 times a day; S4, after the gonads of broodstock mature, artificial induced spawning is carried out, and fertilized eggs are collected for hatching. The present invention makes east star grouper broodstock adapt to artificial environment by manufacturing imitating ecology breeding environment, reduces stress, adopts different feeds for feeding in non-breeding season and breeding season, can meet the ecological factors and nutritional requirements of gonadal development of east star grouper, improves the immunity of east star grouper broodstock and promotes gonadal maturation, improves the quality of sperm and egg, promotes the improvement of fertilization probability, reduces the deformity probability of hatched fry, and improves breeding efficiency.

Description

Technical Field

[0001] The invention relates to the technical field of grouper breeding, and in particular to an ecologically simulated artificial breeding method for grouper. Background Art

[0002] Oriental grouper scientific name Leopard gill perch ( Leopard's Plectropomus ), belonging to the order Perciformes, the family Serranidae, the subfamily Epinephelinae, the genus Gnaphalostreae, and is a warm-water island reef order and carnivorous fish. Its colors include blue, red, brown, and yellow, and its body is thinner than that of ordinary groupers, with a small head. The body and head are from olive to reddish brown, and the head, body, and odd fins are covered with small blue spots. The pectoral fins are red, and there is an inconspicuous dark band on the rear edge of the caudal fin. Its color is bright, with black pupils in its blue eyes, and the fine spots on its body are shaped like stars in the sky, so it is commonly known as "star spot", and "East" because it is produced in the Dongsha Islands. The meat of the Eastern Star Spot is white and tender, rich in nutrition, and is a precious and high-end edible fish. It is deeply favored by consumers and the demand is increasing. At present, although there are methods for artificial breeding of Eastern Star Spot, the maturity of the gonads of male and female broodstock is not uniform during the breeding process, resulting in poor quality of fertilized eggs during the breeding process. At the same time, there are also problems such as low fertilization efficiency and hatching rate of seedlings, and the breeding effect is not ideal. Summary of the invention

[0003] In view of this, the present invention proposes an ecological artificial propagation method for grouper.

[0004] The technical solution of the present invention is achieved in this way:

[0005] A method for artificially breeding grouper fish by simulating ecology comprises the following steps:

[0006] S1. Select 2-3-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding;

[0007] S2. In the non-breeding season, the female broodstock of eastern star grouper are fed with feed A, and the male broodstock of eastern star grouper are fed with feed B; the feed A is prepared by the following raw materials: fish meal, peanut meal, oyster meal, cod liver oil powder, choline chloride, β-carotene, vitamins, extract A, Lactobacillus plantarum, and bifidobacterium; the feed B is prepared by the following raw materials: fish paste, silkworm pupa powder, soybean powder, lecithin, β-carotene, vitamins, extract A, Lactobacillus plantarum, and bifidobacterium; the extract A is prepared by the following raw materials: Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and licorice;

[0008] S3. 2-3 months before reproduction, the female grouper broodstock are fed with fortified feed I, and the male broodstock are fed with fortified feed II; the fortified feed I is made from the following raw materials: fish meal, krill meal, spirulina powder, corn meal, astaxanthin, mixed bacteria, mixed polysaccharides, and extract B; the fortified feed II is made from the following raw materials: minced fish, squid meal, pine needle powder, astaxanthin, mixed bacteria, N-aminoformylglutamic acid, mixed polysaccharides, and extract B; the extract B is made from the following raw materials: patchouli, motherwort, trident, rush, and scutellaria;

[0009] S4. After the gonads of the grouper broodstock mature, artificial induced spawning is performed. After induced spawning, the grouper broodstock are placed in an artificial ecological simulation breeding pond for spawning and fertilization, the fertilized eggs are collected, and the fertilized eggs are placed in a hatching pond for hatching.

[0010] Furthermore, the artificial ecological cultivation pond is: artificial corals are laid on the bottom of the cultivation pond to form an artificial coral reef, and a number of air stones are placed at the bottom for oxygenation.

[0011] Furthermore, the amount of feed A fed each time is 2.0%-3.0% of the body weight of female broodstock, and it is fed 2 to 3 times a day; the amount of feed B fed each time is 2.0%-3.0% of the body weight of male broodstock, and it is fed 2 to 3 times a day; the amount of fortified feed I fed each time is 1.5%-2.0% of the body weight of female broodstock, and it is fed 2 to 3 times a day; the amount of fortified feed II fed each time is 1.5%-2.0% of the body weight of male broodstock, and it is fed 2 to 3 times a day.

[0012] Furthermore, the feed A is prepared by including the following raw materials in parts by weight: 23-28 parts of fish meal, 9-13 parts of peanut meal, 16-20 parts of oyster powder, 6-10 parts of cod liver oil powder, 2-4 parts of choline chloride, 2-3 parts of beta-carotene, 0.1-0.5 parts of vitamins, 9-13 parts of extract A, 0.5-0.9 parts of plant lactobacillus, and 0.5-1.0 parts of bifidobacterium.

[0013] Furthermore, the feed A is prepared by including the following raw materials in parts by weight: 27 parts of fish meal, 10 parts of peanut meal, 17 parts of oyster powder, 7 parts of cod liver oil powder, 2 parts of choline chloride, 3 parts of beta-carotene, 0.4 parts of vitamins, 12 parts of extract A, 0.6 parts of plant lactobacillus, and 0.8 parts of bifidobacterium.

[0014] Furthermore, the feed B is prepared by including the following raw materials in parts by weight: 32-36 parts of fish paste, 20-24 parts of silkworm pupa powder, 12-16 parts of soybean powder, 3-5 parts of lecithin, 3-6 parts of beta-carotene, 0.1-0.2 parts of vitamins, 8-12 parts of extract A, 0.7-1.0 parts of plant lactobacillus, and 0.6-0.8 parts of bifidobacterium.

[0015] Furthermore, the feed B is prepared by including the following raw materials in parts by weight: 33 parts of fish paste, 21 parts of silkworm pupa powder, 13 parts of soybean powder, 3 parts of lecithin, 4 parts of β-carotene, 0.18 parts of vitamins, 11 parts of extract A, 0.9 parts of plant lactobacillus, and 0.7 parts of bifidobacterium.

[0016] Furthermore, the extract A is prepared by including the following raw materials in parts by weight: 12-17 parts of Callicarpa nudiflora, 9-14 parts of Perilla frutescens, 7-13 parts of Cistanche deserticola, and 6-10 parts of Radix Glycyrrhizae; the preparation method of the extract A is: crushing Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae, adding water at a solid-liquid ratio of 1:10-12 g / ml, soaking for 2-3 hours, and then decocting for 90-120 minutes, collecting the extract, concentrating under reduced pressure, drying, and crushing to obtain extract A.

[0017] Furthermore, the fortified feed I comprises the following raw materials in parts by weight: 29-37 parts of fish meal, 17-21 parts of krill meal, 8-13 parts of spirulina powder, 9-12 parts of corn flour, 2-4 parts of astaxanthin, 1.0-1.3 parts of mixed bacterial agent, 1-3 parts of mixed polysaccharides, and 7-13 parts of extract B.

[0018] Furthermore, the fortified feed I comprises the following raw materials in parts by weight: 36 parts of fish meal, 20 parts of krill meal, 9 parts of spirulina powder, 10 parts of corn flour, 2 parts of astaxanthin, 1.2 parts of mixed bacterial agent, 1.3 parts of mixed polysaccharides, and 10 parts of extract B.

[0019] Furthermore, the fortified feed II comprises the following raw materials in parts by weight: 30-34 parts of fish paste, 17-23 parts of squid meal, 12-15 parts of pine needle powder, 7-11 parts of astaxanthin, 2-3 parts of mixed bacterial agent, 0.8-1.3 parts of N-aminoformylglutamic acid, 2-4 parts of mixed polysaccharides, and 9-12 parts of extract B.

[0020] Furthermore, the fortified feed II includes the following raw materials in parts by weight: 30 parts of fish paste, 21 parts of squid powder, 14 parts of pine needle powder, 10 parts of astaxanthin, 2.5 parts of mixed bacterial agent, 1.2 parts of N-aminoformylglutamic acid, 2.5 parts of mixed polysaccharides, and 11 parts of extract B.

[0021] Furthermore, the extract B comprises the following raw materials in parts by weight: 9-12 parts of patchouli, 10-15 parts of motherwort, 11-14 parts of trifoliate bitter orange, 9-12 parts of junctivorous grass, and 7-10 parts of scutellaria baicalensis; the preparation method of the extract B is: crushing the patchouli, motherwort, trifoliate bitter orange, junctivorous grass, and scutellaria baicalensis, adding an ethanol solution with a concentration of 95-98wt% at a solid-liquid ratio of g / ml of 1:6-8, soaking for 2-3 hours, and then extracting at 50-55°C for 3-4 hours, collecting the extract, concentrating under reduced pressure, drying, and crushing to obtain extract B.

[0022] Furthermore, the mixed polysaccharide includes algae polysaccharide, kudzu polysaccharide and liquorice polysaccharide in a mass ratio of 2-5:2-4:1-2; the mixed bacterial agent includes Lactobacillus rhamnosus, Streptococcus thermophilus and Lactobacillus acidophilus in a mass ratio of 4-7:2-4:1-3.

[0023] Furthermore, the artificial induction process is as follows: using HCG and LRH-A3 as oxytocin, per kg of fish body weight, female fish are injected with 600-800 IU of HCG and 3.5-5 μg of LRH-A3, and the dose for male fish is halved, using two injections or one injection.

[0024] Furthermore, during the entire breeding process, the water temperature of female grouper broodstock and male grouper was controlled at 18~32℃, salinity at 25~32‰, pH at 7.8~8.6, and dissolved oxygen content ≥6mg / L.

[0025] Furthermore, during the incubation of fertilized eggs, the water temperature was 24-29°C, the salinity was 25-32‰, the pH was 7.8-8.6, the oxygen concentration was 5-6 mg / L, and the fertilized egg density was 30,000-50,000 eggs / m 3 .

[0026] Furthermore, the feed A, feed B, fortified feed I and fortified feed II are processed into artificial soft granular feed for feeding.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The breeding method of the present invention enables the breeding broodstock of the eastern star grouper to adapt to the environment and reduce stress by creating an ecologically simulated breeding environment, and feeds the female broodstock of the eastern star grouper and the male broodstock with different feeds in the non-breeding season and the breeding season, thereby meeting the ecological factors and nutritional requirements of the gonad development of the eastern star grouper, effectively improving the immunity of the broodstock of the eastern star grouper, promoting the maturation of the gonad, improving the quality of sperm and eggs, and promoting the improvement of the fertilization probability, and at the same time can also reduce the deformity probability of hatched fry and improve the hatching efficiency of fry.

[0029] The invention feeds female broodstock with feed A composed of raw materials such as fish meal, peanut meal, oyster meal, extract A, plant lactobacillus, bifidobacterium and the like in the non-breeding season, and then feeds them with fortified feed I added with mixed bacterial agents, mixed polysaccharides and extract B 3 months before breeding. The two feeds are used in combination to improve the immunity of female broodstock and promote their growth and development, and can also effectively improve the quality of female spawning, and promote the fertilization quality and hatching quality of eggs. The invention uses fish paste, silkworm pupa powder, extract A, plant lactobacillus, bifidobacterium and the like as raw materials as feed B for feeding male broodstock in the non-breeding season, and combines the fortified feed II with mixed bacterial agents, mixed polysaccharides and extract B, which can also enhance the physique of male and female broodstock, promote the growth and development of their gonads, effectively improve the quality of sperm of male broodstock, improve the breeding performance, thereby effectively improving the hatching quality of fry and reducing the deformity of fry. DETAILED DESCRIPTION

[0030] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0031] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0032] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0033] The plant lactobacillus of the present invention is purchased from Shandong Yiwu Biotechnology Co., Ltd., and the effective live bacteria count is 100 billion / gram.

[0034] Bifidobacterium was purchased from Shandong Yiwu Biotechnology Co., Ltd., with an effective viable count of 100 billion / g.

[0035] Lactobacillus rhamnosus was purchased from Shandong Yiwu Biotechnology Co., Ltd., with an effective viable count of 100 billion / g.

[0036] Streptococcus thermophilus was purchased from Jiangsu Jiujia Biotechnology Co., Ltd. with an effective viable count of 10 billion / g.

[0037] Lactobacillus acidophilus was purchased from Shandong Yiwu Biotechnology Co., Ltd., with an effective live bacterial count of 10 billion / g.

[0038] Example 1

[0039] The extract A is prepared by including the following raw materials in parts by weight: 12 parts of Callicarpa nudiflora, 9 parts of Perilla frutescens, 7 parts of Scutellaria baicalensis, and 6 parts of Radix Glycyrrhizae Preparata;

[0040] The preparation method of the extract A is as follows: grind Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae, add water at a solid-liquid ratio of 1:10, soak for 2 hours, and then boil for 90 minutes, collect the extract, concentrate under reduced pressure, dry, and grind to obtain the extract A;

[0041] The extract B is prepared by including the following raw materials in parts by weight: 9 parts of patchouli, 10 parts of motherwort, 11 parts of trifoliate ash, 9 parts of juncis, and 7 parts of scutellaria;

[0042] The preparation method of the extract B is as follows: Patchouli, Leonurus japonicus, Tripterygium wilfordii, Juncus effusus, and Eleutherodactylum sinensis are crushed, and a 95 wt % ethanol solution is added according to a solid-liquid ratio of g / ml of 1:6, and then the mixture is soaked for 2 hours at 50° C. for 3 hours, and the extract is collected, concentrated under reduced pressure, dried, and crushed to obtain the extract B;

[0043] The feed A is prepared by including the following raw materials in parts by weight: 23 parts of fish meal, 9 parts of peanut meal, 16 parts of oyster meal, 6 parts of cod liver oil powder, 2 parts of choline chloride, 2 parts of beta-carotene, 0.1 parts of vitamins, 9 parts of extract A, 0.5 parts of plant lactobacillus, and 0.5 parts of bifidobacterium;

[0044] The feed B is prepared by including the following raw materials in parts by weight: 32 parts of fish paste, 20 parts of silkworm pupa powder, 12 parts of soybean powder, 3 parts of lecithin, 3 parts of β-carotene, 0.1 parts of vitamins, 8 parts of extract A, 0.7 parts of plant lactobacillus, and 0.6 parts of bifidobacterium;

[0045] The fortified feed I comprises the following raw materials in parts by weight: 29 parts of fish meal, 17 parts of krill meal, 8 parts of spirulina powder, 9 parts of corn flour, 2 parts of astaxanthin, 1.0 part of mixed bacterial agent, 1 part of mixed polysaccharide, and 7 parts of extract B;

[0046] The fortified feed II comprises the following raw materials in parts by weight: 30 parts of fish paste, 17 parts of squid powder, 12 parts of pine needle powder, 7 parts of astaxanthin, 2 parts of mixed bacterial agent, 0.8 parts of N-aminoformylglutamic acid, 2 parts of mixed polysaccharides, and 9 parts of extract B;

[0047] The mixed polysaccharide includes algae polysaccharide, kudzu polysaccharide and liquorice polysaccharide in a mass ratio of 2:2:1; the mixed bacterial agent includes Lactobacillus rhamnosus, Streptococcus thermophilus and Lactobacillus acidophilus in a mass ratio of 4:2:1.

[0048] The ecological artificial breeding method of grouper fish of the present embodiment comprises the following steps:

[0049] S1. Select 2-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 23°C, the salinity is 25‰, the pH is 7.8, and the dissolved oxygen content is ≥6mg / L;

[0050] S2. In the non-breeding season, the female grouper broodstock are fed with feed A, and the male grouper broodstock are fed with feed B; the feed A is fed in an amount of 2.0% of the body weight of the female broodstock, and is fed twice a day; the feed B is fed in an amount of 2.0% of the body weight of the male broodstock, and is fed twice a day;

[0051] S3. Three months before breeding, the female grouper broodstock were fed with fortified feed I, and the male broodstock were fed with fortified feed II; the fortified feed I was fed at a rate of 1.5% of the body weight of the female broodstock twice a day; the fortified feed II was fed at a rate of 1.5% of the body weight of the male broodstock twice a day;

[0052] S4. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out, using HCG and LRH-A3 as oxytocin. For each kg of fish body weight, female fish are injected with 600 IU of HCG and 3.5 μg of LRH-A3, and the dose for male fish is halved, using a double injection or a single injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 24°C, the salinity is 25‰, the pH is 7.8, the oxygen concentration is 5 mg / L, and the fertilized egg density is 30,000 / m 3 .

[0053] Example 2

[0054] The extract A is prepared by including the following raw materials in parts by weight: 17 parts of Callicarpa nudiflora, 14 parts of Perilla frutescens, 13 parts of Cnidium monnieri, and 10 parts of Radix Glycyrrhizae Preparata;

[0055] The preparation method of the extract A is as follows: grind Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae, add water at a solid-liquid ratio of 1:12, soak for 3 hours, and then boil for 120 minutes, collect the extract, concentrate under reduced pressure, dry, and grind to obtain the extract A;

[0056] The extract B is prepared by including the following raw materials in parts by weight: 12 parts of patchouli, 15 parts of motherwort, 14 parts of trifoliate ash, 12 parts of juncis, and 10 parts of scutellaria;

[0057] The preparation method of the extract B is as follows: Patchouli, Leonurus japonicus, Tripterygium wilfordii, Juncus effusus, and Eleutherodactylum sinensis are crushed, and a 98 wt % ethanol solution is added according to a solid-liquid ratio of g / ml of 1:8, and then the mixture is soaked for 3 hours at 55° C. for 4 hours, and the extract is collected, concentrated under reduced pressure, dried, and crushed to obtain the extract B;

[0058] The feed A is prepared by including the following raw materials in parts by weight: 28 parts of fish meal, 13 parts of peanut meal, 20 parts of oyster meal, 10 parts of cod liver oil powder, 4 parts of choline chloride, 3 parts of beta-carotene, 0.5 parts of vitamins, 13 parts of extract A, 0.9 parts of plant lactobacillus, and 1.0 parts of bifidobacterium;

[0059] The feed B is prepared by including the following raw materials in parts by weight: 36 parts of fish paste, 24 parts of silkworm pupa powder, 16 parts of soybean powder, 5 parts of lecithin, 6 parts of β-carotene, 0.2 parts of vitamins, 12 parts of extract A, 1.0 parts of plant lactobacillus, and 0.8 parts of bifidobacterium;

[0060] The fortified feed I comprises the following raw materials in parts by weight: 37 parts of fish meal, 21 parts of krill meal, 13 parts of spirulina powder, 12 parts of corn flour, 4 parts of astaxanthin, 1.3 parts of mixed bacterial agent, 3 parts of mixed polysaccharide, and 13 parts of extract B;

[0061] The fortified feed II comprises the following raw materials in parts by weight: 34 parts of fish paste, 23 parts of squid powder, 15 parts of pine needle powder, 11 parts of astaxanthin, 3 parts of mixed bacterial agent, 1.3 parts of N-aminoformylglutamic acid, 4 parts of mixed polysaccharides, and 12 parts of extract B;

[0062] The mixed polysaccharide includes algae polysaccharide, kudzu root polysaccharide and liquorice polysaccharide in a mass ratio of 5:4:2; the mixed bacterial agent includes rhamnosus lactobacillus, thermophilic streptococcus and acidophilus lactobacillus in a mass ratio of 7:4:3.

[0063] The ecological artificial breeding method of grouper fish of the present embodiment comprises the following steps:

[0064] S1. Select 3-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 28°C, the salinity is 27‰, the pH is 8.4, and the dissolved oxygen content is ≥6mg / L;

[0065] S2. In the non-breeding season, the female grouper broodstock are fed with feed A, and the male grouper broodstock are fed with feed B; the feed A is fed in an amount of 3.0% of the body weight of the female broodstock, and is fed twice a day; the feed B is fed in an amount of 3.0% of the body weight of the male broodstock, and is fed twice a day;

[0066] S3. Three months before breeding, the female grouper broodstock were fed with fortified feed I, and the male broodstock were fed with fortified feed II; the fortified feed I was fed at a rate of 2.0% of the body weight of the female broodstock twice a day; the fortified feed II was fed at a rate of 2.0% of the body weight of the male broodstock twice a day;

[0067] S4. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out, using HCG and LRH-A3 as oxytocin. For every kg of fish body weight, female fish are injected with 800 IU of HCG and 5 μg of LRH-A3, and the dose for male fish is halved, using a double injection or a single injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 28°C, the salinity is 30‰, the pH is 8.4, the oxygen concentration is 6 mg / L, and the fertilized egg density is 50,000 / m 3 .

[0068] Example 3

[0069] The extract A is prepared by including the following raw materials in parts by weight: 15 parts of Callicarpa nudiflora, 10 parts of Perilla frutescens, 9 parts of Scutellaria baicalensis, and 8 parts of Radix Glycyrrhizae Preparata;

[0070] The preparation method of the extract A is as follows: grind Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae Radix, add water at a solid-liquid ratio of 1:10, soak for 3 hours, and then boil for 120 minutes, collect the extract, concentrate under reduced pressure, dry, and grind to obtain the extract A;

[0071] The extract B is prepared by including the following raw materials in parts by weight: 11 parts of patchouli, 13 parts of motherwort, 12 parts of trifoliate bitter orange, 10 parts of juncis effusus, and 9 parts of scutellaria baicalensis;

[0072] The preparation method of the extract B is as follows: Patchouli, Leonurus japonicus, Tripterygium wilfordii, Juncus effusus, and Eleutherodactylum sinensis are crushed, and a 96 wt % ethanol solution is added according to a solid-liquid ratio of g / ml of 1:8, and then the mixture is soaked for 3 hours at 55° C. for 4 hours, and the extract is collected, concentrated under reduced pressure, dried, and crushed to obtain the extract B;

[0073] The feed A is prepared by including the following raw materials in parts by weight: 24 parts of fish meal, 13 parts of peanut meal, 18 parts of oyster meal, 8 parts of cod liver oil powder, 3 parts of choline chloride, 2.5 parts of beta-carotene, 0.3 parts of vitamins, 11 parts of extract A, 0.8 parts of plant lactobacillus, and 0.7 parts of bifidobacterium;

[0074] The feed B is prepared by including the following raw materials in parts by weight: 35 parts of fish paste, 23 parts of silkworm pupa powder, 14 parts of soybean powder, 4 parts of lecithin, 5 parts of β-carotene, 0.15 parts of vitamins, 10 parts of extract A, 0.8 parts of plant lactobacillus, and 0.8 parts of bifidobacterium;

[0075] The fortified feed I comprises the following raw materials in parts by weight: 32 parts of fish meal, 18 parts of krill meal, 12 parts of spirulina powder, 11 parts of corn flour, 3 parts of astaxanthin, 1.3 parts of mixed bacterial agent, 2 parts of mixed polysaccharide, and 12 parts of extract B;

[0076] The fortified feed II comprises the following raw materials in parts by weight: 32 parts of fish paste, 19 parts of squid powder, 12 parts of pine needle powder, 8 parts of astaxanthin, 3 parts of mixed bacterial agent, 1.1 parts of N-aminoformylglutamic acid, 3 parts of mixed polysaccharides, and 10 parts of extract B;

[0077] The mixed polysaccharide includes algae polysaccharide, kudzu root polysaccharide and liquorice polysaccharide in a mass ratio of 4:2.5:1; the mixed bacterial agent includes rhamnosus lactobacillus, thermophilic streptococcus and acidophilus lactobacillus in a mass ratio of 5:2.4:2.5.

[0078] The ecological artificial breeding method of grouper fish of the present embodiment comprises the following steps:

[0079] S1. Select 2-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 25°C, the salinity is 26‰, the pH is 7.8, and the dissolved oxygen content is ≥6mg / L;

[0080] S2. In the non-breeding season, the female grouper broodstock are fed with feed A, and the male grouper broodstock are fed with feed B; the feed A is fed in an amount of 2.5% of the body weight of the female broodstock, and is fed twice a day; the feed B is fed in an amount of 2.5% of the body weight of the male broodstock, and is fed twice a day;

[0081] S3. Three months before breeding, the female grouper broodstock were fed with fortified feed I, and the male broodstock were fed with fortified feed II; the fortified feed I was fed at a rate of 1.6% of the body weight of the female broodstock twice a day; the fortified feed II was fed at a rate of 1.8% of the body weight of the male broodstock twice a day;

[0082] S4. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out, using HCG and LRH-A3 as oxytocin. The female fish is injected with 700 IU of HCG and 4.5 μg of LRH-A3 per kg of fish body weight, and the dose of male fish is halved, using a single injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 28°C, the salinity is 27‰, the pH is 8.0, the oxygen concentration is 5 mg / L, and the fertilized egg density is 40,000 / m 3 .

[0083] Example 4

[0084] The extract A is prepared by including the following raw materials in parts by weight: 16 parts of Callicarpa nudiflora, 12 parts of Perilla frutescens, 7 parts of Scutellaria baicalensis, and 9 parts of Radix Glycyrrhizae Preparata;

[0085] The preparation method of the extract A is as follows: grind Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae, add water at a solid-liquid ratio of 1:11, soak for 2 hours, and then boil for 100 minutes, collect the extract, concentrate under reduced pressure, dry, and grind to obtain the extract A;

[0086] The extract B is prepared by including the following raw materials in parts by weight: 10 parts of patchouli, 11 parts of motherwort, 13 parts of trifoliate ash, 11 parts of juncea, and 8 parts of scutellaria baicalensis;

[0087] The preparation method of the extract B is as follows: Patchouli, Leonurus japonicus, Tripterygium wilfordii, Juncus effusus, and Eleutherodactylum sinensis are crushed, and a 97 wt % ethanol solution is added according to a solid-liquid ratio of g / ml of 1:7, and then the mixture is soaked for 2.5 hours at 50° C. for 3.5 hours, and the extract is collected, concentrated under reduced pressure, dried, and crushed to obtain the extract B;

[0088] The feed A is prepared by including the following raw materials in parts by weight: 27 parts of fish meal, 10 parts of peanut meal, 17 parts of oyster meal, 7 parts of cod liver oil powder, 2 parts of choline chloride, 3 parts of beta-carotene, 0.4 parts of vitamins, 12 parts of extract A, 0.6 parts of plant lactobacillus, and 0.8 parts of bifidobacterium;

[0089] The feed B is prepared by including the following raw materials in parts by weight: 33 parts of fish paste, 21 parts of silkworm pupa powder, 13 parts of soybean powder, 3 parts of lecithin, 4 parts of β-carotene, 0.18 parts of vitamins, 11 parts of extract A, 0.9 parts of plant lactobacillus, and 0.7 parts of bifidobacterium;

[0090] The fortified feed I comprises the following raw materials in parts by weight: 36 parts of fish meal, 20 parts of krill meal, 9 parts of spirulina powder, 10 parts of corn flour, 2 parts of astaxanthin, 1.2 parts of mixed bacterial agent, 1.3 parts of mixed polysaccharides, and 10 parts of extract B;

[0091] The fortified feed II comprises the following raw materials in parts by weight: 30 parts of fish paste, 21 parts of squid powder, 14 parts of pine needle powder, 10 parts of astaxanthin, 2.5 parts of mixed bacterial agent, 1.2 parts of N-aminoformylglutamic acid, 2.5 parts of mixed polysaccharides, and 11 parts of extract B;

[0092] The mixed polysaccharide includes algae polysaccharide, kudzu root polysaccharide and liquorice polysaccharide in a mass ratio of 3:3:1.8; the mixed bacterial agent includes rhamnosus lactobacillus, thermophilic streptococcus and acidophilus lactobacillus in a mass ratio of 6:3:2.

[0093] The ecological artificial breeding method of grouper fish of the present embodiment comprises the following steps:

[0094] S1. Select 3-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 26°C, the salinity is 28‰, the pH is 8.2, and the dissolved oxygen content is ≥6mg / L;

[0095] S2. In the non-breeding season, the female grouper broodstock are fed with feed A, and the male grouper broodstock are fed with feed B; the feed A is fed at an amount of 2.8% of the body weight of the female broodstock twice a day; the feed B is fed at an amount of 2.6% of the body weight of the male broodstock twice a day;

[0096] S3. Three months before breeding, the female grouper broodstock were fed with fortified feed I, and the male broodstock were fed with fortified feed II; the fortified feed I was fed at a rate of 1.5% of the body weight of the female broodstock twice a day; the fortified feed II was fed at a rate of 1.7% of the body weight of the male broodstock twice a day;

[0097] S4. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out, using HCG and LRH-A3 as oxytocin. The female fish is injected with 650 IU of HCG and 4 μg of LRH-A3 per kg of fish body weight, and the dose of male fish is halved, using a second injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 26°C, the salinity is 28‰, the pH is 8.0, the oxygen concentration is 6 mg / L, and the fertilized egg density is 50,000 / m 3 .

[0098] Comparative Example 1

[0099] This comparative example is adjusted on the basis of Example 4. During the entire breeding process, only the female broodstock feed A and only the male broodstock feed B are fed, and the fortified feed I and fortified feed II are not fed. The rest is consistent with Example 4.

[0100] The ecological artificial breeding method of grouper in this comparative example comprises the following steps:

[0101] S1. Select 3-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 26°C, the salinity is 28‰, the pH is 8.2, and the dissolved oxygen content is ≥6mg / L;

[0102] S2. During the non-breeding season and 2-3 months before breeding, the female grouper broodstock were fed with feed A and the male grouper broodstock were fed with feed B; the feed A was fed at an amount of 2.8% of the body weight of the female broodstock twice a day; the feed B was fed at an amount of 2.6% of the body weight of the male broodstock twice a day;

[0103] S3. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out. HCG and LRH-A3 are used as oxytocin. The female fish are injected with 650 IU of HCG and 4 μg of LRH-A3 per kg of fish body weight, and the dose of male fish is halved, with a second injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 26°C, the salinity is 28‰, the pH is 8.0, the oxygen concentration is 6 mg / L, and the fertilized egg density is 50,000 / m 3 .

[0104] Comparative Example 2

[0105] This comparative example is adjusted on the basis of Example 4. During the entire breeding process, only the female broodstock are fed with fortified feed I, and only the male broodstock are fed with fortified feed II. Feed A and feed B are not fed. The rest is consistent with Example 4.

[0106] The ecological artificial breeding method of grouper in this comparative example comprises the following steps:

[0107] S1. Select 3-year-old disease-free female and male grouper broodstock, and place the female and male broodstock in artificial ecological breeding ponds for breeding respectively; the artificial ecological breeding ponds are: artificial corals are laid at the bottom of the breeding pond to form artificial coral reefs, and a number of air stones are placed at the bottom for oxygenation; during the whole breeding process, the water temperature is controlled at 26°C, the salinity is 28‰, the pH is 8.2, and the dissolved oxygen content is ≥6mg / L;

[0108] S2. During the non-breeding season and 2-3 months before breeding, the female grouper broodstock are fed with fortified feed I, and the male broodstock are fed with fortified feed II; the fortified feed I is fed at a rate of 1.5% of the body weight of the female broodstock twice a day; the fortified feed II is fed at a rate of 1.7% of the body weight of the male broodstock twice a day;

[0109] S3. After the gonads of the grouper broodstock mature, artificial induction of spawning is carried out. HCG and LRH-A3 are used as oxytocin. The female fish are injected with 650 IU of HCG and 4 μg of LRH-A3 per kg of fish body weight, and the dose of male fish is halved, with a second injection. After induction of spawning, the grouper broodstock are placed in an artificial ecological breeding pond for spawning and fertilization. The fertilized eggs are collected and placed in an incubation pond for incubation. During the incubation of the fertilized eggs, the water temperature is 26°C, the salinity is 28‰, the pH is 8.0, the oxygen concentration is 6 mg / L, and the fertilized egg density is 50,000 / m 3 .

[0110] Comparative Example 3

[0111] This comparative example is adjusted on the basis of Example 4, wherein neither Feed A nor Feed B contains Extract A, and the rest is consistent with Example 4.

[0112] The feed A of this comparative example is prepared by including the following raw materials in parts by weight: 39 parts of fish meal, 10 parts of peanut meal, 17 parts of oyster meal, 7 parts of cod liver oil powder, 2 parts of choline chloride, 3 parts of β-carotene, 0.4 parts of vitamins, 0.6 parts of plant lactobacillus, and 0.8 parts of bifidobacterium;

[0113] The feed B is prepared from the following raw materials in parts by weight: 44 parts of fish paste, 21 parts of silkworm pupa powder, 13 parts of soybean powder, 3 parts of lecithin, 4 parts of beta-carotene, 0.18 parts of vitamins, 0.9 parts of plant lactobacillus, and 0.7 parts of bifidobacterium.

[0114] Comparative Example 4

[0115] This comparative example is adjusted on the basis of Example 4. The fortified feed I and fortified feed II do not contain extract B and mixed polysaccharides. The rest is consistent with Example 4.

[0116] The fortified feed I of this comparative example comprises the following raw materials in parts by weight: 41 parts of fish meal, 25 parts of krill meal, 10.3 parts of spirulina powder, 10 parts of corn flour, 2 parts of astaxanthin, and 1.2 parts of mixed bacterial agent;

[0117] The fortified feed II comprises the following raw materials in parts by weight: 36 parts of fish paste, 28.5 parts of squid powder, 14 parts of pine needle powder, 10 parts of astaxanthin, 2.5 parts of mixed bacterial agent, and 1.2 parts of N-aminoformylglutamic acid.

[0118] The grouper was bred according to the breeding methods of Examples 1-4 and Comparative Examples 1-4, with 150 female broodstock in each group. The fertilization rate, hatching rate and fry deformity rate during the breeding process were counted, and the results were averaged, as shown in Table 1 below.

[0119] Table 1

[0120]

[0121] As can be seen from the table, compared with Example 4, in Comparative Example 1, female broodstock were not fed with fortified feed I and male broodstock were not fed with fortified feed II before breeding. Compared with Example 4, Comparative Example 2 did not feed female broodstock with feed A and did not feed male broodstock with feed B in the non-breeding season, which affected the gonadal maturation of male and female broodstock of the eastern star grouper, affected the fertilization, and also caused the quality of fertilized eggs to decrease. Compared with Example 4, the feed A and feed B of Comparative Example 3 did not contain extract A, and the fortified feed I and fortified feed II of Comparative Example 4 did not contain extract B and mixed polysaccharides, which also reduced the reproductive efficiency of male and female broodstock of the eastern star grouper. The results show that the ecological artificial breeding method of the present invention can promote the breeding efficiency of the eastern star grouper broodstock, especially through the synergistic combination of the specially formulated feed A, feed B, fortified feed I and fortified feed II of the present invention, the gonadal maturation of the male and female eastern star grouper broodstock can be effectively promoted, the maturity of the eggs can be ensured, and the vitality and quality of the eggs and sperms can also be ensured, the fertilization rate and hatching rate can be effectively improved, and the deformity rate of the fry can be reduced, and the breeding efficiency is high. In particular, the extract A prepared from Callicarpa nudiflora, Perilla frutescens, Caulis ternata and Glycyrrhiza uralensis, the extract B composed of Patchouli, Leonurus japonicus, Tripterygium wilfordii, Juncus wicker and Eleutherodactylum sinensis, and the mixed polysaccharide composed of Euglena polysaccharide, Pueraria polysaccharide and Glycyrrhiza uralensis can improve the immunity of the male and female eastern star grouper broodstock, enhance the physique of the male and female broodstock, promote the growth and development of their gonads, effectively improve the quality of the eggs and sperms of the male and female broodstock, improve the breeding performance, thereby effectively improving the hatching quality of the fry and reducing the deformity of the fry.

[0122] 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 artificial breeding of grouper by simulating ecology, characterized in that: The following steps are involved: S1. Select 2-3-year-old disease-free female and male grouper broodstock, and place the female and male grouper broodstock in artificial ecological breeding ponds for breeding; S2. In the non-breeding season, the female broodstock of eastern star grouper are fed with feed A, and the male broodstock of eastern star grouper are fed with feed B; the feed A comprises the following raw materials: fish meal, peanut meal, oyster meal, cod liver oil powder, choline chloride, β-carotene, vitamins, extract A, Lactobacillus plantarum, and bifidobacterium; the feed B comprises the following raw materials: fish paste, silkworm pupa powder, soybean powder, lecithin, β-carotene, vitamins, extract A, Lactobacillus plantarum, and bifidobacterium; the extract A comprises the following raw materials: Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and licorice; S3. 2-3 months before reproduction, feed the female grouper broodstock with fortified feed I and feed the male grouper broodstock with fortified feed II; the fortified feed I comprises the following raw materials: fish meal, krill meal, spirulina powder, corn meal, astaxanthin, mixed bacteria, mixed polysaccharides, and extract B; the fortified feed II comprises the following raw materials: fish paste, squid meal, pine needle powder, astaxanthin, mixed bacteria, N-aminoformylglutamic acid, mixed polysaccharides, and extract B; the extract B comprises the following raw materials: patchouli, motherwort, trident, rush, and scutellaria; S4. After the gonads of the female and male grouper broodstock mature, artificial induced spawning is carried out. After induced spawning, the female and male grouper broodstock are placed in an artificial ecological simulation breeding pond for spawning and fertilization, the fertilized eggs are collected, and the fertilized eggs are placed in a hatching pond for hatching.

2. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The artificial ecological breeding pool is as follows: artificial corals are laid on the bottom of the breeding pool to form an artificial coral reef, and a plurality of air bubbles are placed on the bottom to increase oxygen.

3. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The amount of feed A fed each time is 2.0%-3.0% of the body weight of female grouper broodstock, and it is fed 2 to 3 times a day; the amount of feed B fed each time is 2.0%-3.0% of the body weight of male grouper broodstock, and it is fed 2 to 3 times a day; the amount of fortified feed I fed each time is 1.5%-2.0% of the body weight of female grouper broodstock, and it is fed 2 to 3 times a day; the amount of fortified feed II fed each time is 1.5%-2.0% of the body weight of male grouper broodstock, and it is fed 2 to 3 times a day.

4. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The feed A comprises the following raw materials in parts by weight: 23-28 parts of fish meal, 9-13 parts of peanut meal, 16-20 parts of oyster powder, 6-10 parts of cod liver oil powder, 2-4 parts of choline chloride, 2-3 parts of beta-carotene, 0.1-0.5 parts of vitamins, 9-13 parts of extract A, 0.5-0.9 parts of plant lactobacillus, and 0.5-1.0 parts of bifidobacteria; the feed B comprises the following raw materials in parts by weight: 32-36 parts of fish paste, 20-24 parts of silkworm pupa powder, 12-16 parts of soybean powder, 3-5 parts of lecithin, 3-6 parts of beta-carotene, 0.1-0.2 parts of vitamins, 8-12 parts of extract A, 0.7-1.0 parts of plant lactobacillus, and 0.6-0.8 parts of bifidobacteria.

5. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The extract A comprises the following raw materials in parts by weight: 12-17 parts of Callicarpa nudiflora, 9-14 parts of Perilla frutescens, 7-13 parts of Cistanche deserticola, and 6-10 parts of Radix Glycyrrhizae; the preparation method of the extract A is: crush Callicarpa nudiflora, Perilla frutescens, Cistanche deserticola, and Radix Glycyrrhizae, add water at a solid-liquid ratio of 1:10-12, soak for 2-3 hours, and then boil for 90-120 minutes, collect the extract, concentrate under reduced pressure, dry, and crush to obtain extract A.

6. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The fortified feed I comprises the following raw materials in parts by weight: 29-37 parts of fish meal, 17-21 parts of krill meal, 8-13 parts of spirulina powder, 9-12 parts of corn flour, 2-4 parts of astaxanthin, 1.0-1.3 parts of mixed bacterial agent, 1-3 parts of mixed polysaccharides, and 7-13 parts of extract B; the fortified feed II comprises the following raw materials in parts by weight: 30-34 parts of fish paste, 17-23 parts of squid meal, 12-15 parts of pine needle powder, 7-11 parts of astaxanthin, 2-3 parts of mixed bacterial agent, 0.8-1.3 parts of N-aminoformylglutamic acid, 2-4 parts of mixed polysaccharides, and 9-12 parts of extract B.

7. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The extract B comprises the following raw materials in parts by weight: 9-12 parts of patchouli, 10-15 parts of motherwort, 11-14 parts of trifoliate cyperus, 9-12 parts of juncchus effusus, and 7-10 parts of scutellaria baicalensis. The preparation method of the extract B is as follows: the patchouli, motherwort, trifoliate cyperus, juncchus effusus, and scutellaria baicalensis are crushed, and an ethanol solution with a concentration of 95-98wt% is added thereto at a solid-liquid ratio of g / ml of 1:6-8, and the mixture is immersed for 2-3 hours, and then the mixture is extracted at 50-55°C for 3-4 hours, and the extract is collected, concentrated under reduced pressure, dried, and crushed to obtain the extract B.

8. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The mixed polysaccharide comprises algae polysaccharide, kudzu root polysaccharide and liquorice polysaccharide in a mass ratio of 2-5:2-4:1-2; the mixed bacterial agent comprises rhamnosus lactobacillus, thermophilic streptococcus and acidophilus lactobacillus in a mass ratio of 4-7:2-4:1-3.

9. The ecological artificial breeding method of grouper according to claim 1, characterized in that: The artificial induction process is as follows: using HCG and LRH-A3 as oxytocin, per kg of fish body weight, female grouper broodstock are injected with 600-800 IU of HCG and 3.5-5 μg of LRH-A3, and the dose for male grouper broodstock is halved, and two injections or one injection is used.

10. The ecological artificial breeding method of grouper according to claim 1, characterized in that: During the entire breeding process, the water temperature of the female and male grouper broodstock was controlled at 18-32°C, salinity at 25-32‰, pH at 7.8-8.6, and dissolved oxygen at ≥6mg / L; during the hatching of fertilized eggs, the water temperature was controlled at 24-29°C, salinity at 25-32‰, pH at 7.8-8.6, oxygen concentration at 5-6mg / L, and fertilized egg density at 30,000-50,000 / m 3 .

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