Artificial breeding method of fir and tiger hybrid spots

Abstract

The invention provides an artificial breeding method of fir and tiger hybrid spots, and relates to the field of aquaculture. The artificial breeding method of the fir-tiger hybrid spots comprises the following steps: (1) parent selection; (2) parent breeding: starting to feed female parent fish feed I and male parent fish feed II three months before breeding; the feed I comprises the following raw materials: fish meal, soybean meal, corn flour, astaxanthin, beta-glucan, a complex microbial inoculant and a compound A; the feed II comprises the following raw materials: fish meal, squid powder, dunaliella salina powder, astaxanthin, rosmarinic acid, a complex microbial inoculant and a compound B; the composite microbial agent is prepared from the following raw materials: acetobacter xylinum, bacillus pumilus and bacillus natto; the compound A comprises the following raw materials: dendrobium officinale, gynostemma pentaphylla, ramie roots, aralia elata leaves and perilla seeds; the compound B comprises the following raw materials: hippocampus, cistanche, acanthopanax, water chestnut hull and moringa seed; (3) fertilization and incubation to obtain larva fish. The breeding method has the advantages of high egg laying amount, high fertilization rate and high hatching rate, and fry malformation is reduced.

Description

Technical Field

[0001] The invention relates to the field of aquaculture, and in particular to an artificial breeding method for hybrid spotted mullet. Background Art

[0002] Clearwater Grouper (Spruce Grouper) Epinephelus Polyphekadion The meat is delicious and has strong resistance to adversity. Epinephelus Fuscoguttatus It has a wide temperature adaptability, fast growth rate, delicious meat and high nutritional value. The hybrid species of Shanhu hybrid grouper is a hybrid bred with the clearwater grouper as the female parent and the brown-spotted grouper as the male parent.

[0003] Traditional breeding of hybrid pine tiger spotters results in a long rearing cycle, is prone to disease, and reduces the survival rate of adult fish. Artificial breeding of improved species is an effective way to address these issues at the source. Traditional breeding of hybrid pine tiger spotters results in limited egg production, low fertilization and hatching rates, and some fry are deformed. Outdoor breeding is even more challenging. Therefore, there is an urgent need to develop an artificial breeding method for hybrid pine tiger spotters. Summary of the Invention

[0004] In view of this, the present invention proposes an artificial breeding method for hybrid spotted pine tiger to solve the above problems.

[0005] The technical solution of the present invention is achieved as follows: an artificial breeding method of cedar tiger hybrid spot, comprising the following steps: (1) Parent selection: Select healthy 3-4-year-old freshwater grouper as female parent; select healthy 1-2-year-old brown-spotted grouper as male parent; (2) Parent cultivation: 3 months before breeding, the female broodstock were fed with fortified feed I and the male broodstock were fed with fortified feed II; the feed I included the following raw materials: fish meal, soybean meal, corn meal, astaxanthin, β-glucan, compound bacteria agent, and compound A; the feed II included the following raw materials: fish meal, squid meal, salt algae meal, astaxanthin, rosmarinic acid, compound bacteria agent, and compound B; the compound bacteria agent included the following raw materials: Acetobacter xylinum, Bacillus pumilus, and Bacillus natto; the compound A included the following raw materials: Dendrobium officinale, Gynostemma pentaphyllum, ramie root, Aralia oleifera leaf, and Perilla seed; the compound B included the following raw materials: seahorse, Cistanche deserticola, Acanthopanax senticosus, water chestnut shell, and Moringa seed; (3) Fertilization and hatching: When the parent fish reach sexual maturity, they will naturally lay eggs and fertilize them. The fertilized eggs will be transferred to an incubator for hatching to obtain fry.

[0006] Furthermore, the environment for the parent cultivation is: select an open-air high-level pond with a water depth of 2-4m for cultivation, and a cultivation density of 3-5 tails / m 3 .

[0007] Furthermore, the feed I comprises the following raw materials in parts by weight: 25-35 parts of fish meal, 15-25 parts of soybean meal, 10-20 parts of corn flour, 3-8 parts of astaxanthin, 8-12 parts of β-glucan, 2-4 parts of composite bacterial agent, and 15-20 parts of complex A; the daily feeding amount of the feed I is 2%-3% of the weight of the parent, and it is fed 2-3 times a day.

[0008] Furthermore, the feed II comprises the following raw materials in parts by weight: 20-30 parts of fish meal, 20-30 parts of squid meal, 10-20 parts of salt algae powder, 8-12 parts of astaxanthin, 3-8 parts of rosmarinic acid, 3-8 parts of composite bacterial agent, and 13-18 parts of complex B; the daily feeding amount of the feed II is 1%-2% of the weight of the parent, and it is fed 2-3 times a day.

[0009] Furthermore, the compound A comprises the following raw materials in parts by weight: 25-35 parts of Dendrobium officinale, 20-30 parts of Gynostemma pentaphyllum, 15-25 parts of ramie root, 10-20 parts of Aralia dasyphylla leaves, and 8-12 parts of Perilla seeds.

[0010] Furthermore, the compound B comprises the following raw materials in parts by weight: 20-30 parts of seahorse, 20-30 parts of Cistanche deserticola, 15-25 parts of Acanthopanax senticosus, 10-20 parts of water chestnut shell, and 10-20 parts of Moringa seeds.

[0011] Furthermore, the mass ratio of Acetobacter xylinum, Bacillus pumilus and Bacillus natto in the composite bacterial agent is 4-6:3-4:1-2.

[0012] Furthermore, during the parent cultivation process, 24 hours before the rainstorm, the enhancer was added at a rate of 0.4-0.6g / m 2 The enhancer includes the following raw materials in parts by weight: 25-35 parts of Aralia dasyphylla extract, 4-6 parts of L-valine, 8-12 parts of thiamine, 10-20 parts of disodium ethylenediaminetetraacetic acid, 15-25 parts of magnesium sulfate, and 15-25 parts of calcium dihydrogen phosphate.

[0013] Furthermore, before adding the fertilized eggs, seawater is placed in the incubator, pullulan is added, and plasma treatment is performed at 60-70W and 40-50kHz for 2-4 minutes. The amount of pullulan added is 0.05%-0.1% of the seawater.

[0014] Furthermore, after adding the fertilized eggs, the magnetization treatment is performed at 50-80 Gs for 20-30 minutes.

[0015] Furthermore, during the incubation process, the water temperature is 22-28°C, the salinity is 28‰~30‰, the pH value is 7.5-8.5, the oxygen concentration is 4-6 mg / L, and the fertilized egg density is 30,000-40,000 eggs / m 3 .

[0016] Compared with the prior art, the present invention has the following beneficial effects: (1) The artificial breeding method of the hybrid spot of the pine tiger of the present invention is used. The single female fish has a high egg production, high fertilization rate and high hatching rate, and minimizes the deformity of the fry. Among them, the present invention uses the compound A prepared by combining ramie root, Aralia dasyphylla leaves with Dendrobium officinale, Gynostemma pentaphyllum, and Perilla seeds, which is beneficial to the development and maturation of the gonads of female fish, increases the number of eggs laid and the quality of eggs laid, improves the fertilization rate and hatching rate, and reduces the deformity rate. The present invention uses Acanthopanax senticosus, water chestnut shell, combined with seahorse, Cistanche deserticola, and Moringa seeds to prepare the compound B, which is beneficial to the development and maturation of the gonads of male fish, improves the quality of their semen, significantly improves the fertilization rate and hatching rate, and significantly reduces the deformity rate. The present invention uses Acetobacter xylinum, Bacillus pumilus, and Bacillus natto to prepare feed I and feed II, which further promote the development of the gonads of male and female fish and improve the quality of fertilized eggs. At the same time, the feed I and feed II prepared by the present invention also effectively improve the immunity and anti-stress response ability of the parents, and enhance their breeding ability.

[0017] (2) The present invention adds a fortifier prepared by combining gentian root extract with L-valine, thiamine, disodium ethylenediaminetetraacetic acid, magnesium sulfate, and calcium dihydrogen phosphate before heavy rain during the parent breeding process, which is beneficial to reducing the adverse effects of heavy rain on the gonadal development of the parents.

[0018] (3) The present invention selects an open-air high-level pond for parent cultivation. Combining the method of the present invention can not only overcome the adverse effects of weather, but also help improve the parents' ability to resist stress and enhance their breeding quality. DETAILED DESCRIPTION

[0019] In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention.

[0020] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.

[0021] Unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources.

[0022] The Acetobacter xylinum, Bacillus pumilus and Bacillus natto used in the examples of the present invention are all commercially available strains.

[0023] Example 1 An artificial breeding method for hybrid pine tiger spot, comprising the following steps: (1) Parent selection: Select healthy freshwater groupers aged 3-4 years as female parents; select healthy brown-spotted groupers aged 1-2 years as male parents.

[0024] (2) Parent cultivation: Cultivation environment: Choose an open-air high-level pond with a water depth of 2-4m for cultivation, with a cultivation density of 3-5 fish / m 3 ; Three months before breeding, the female broodstock were fed fortified feed I and the male broodstock were fed fortified feed II. Feed I included the following ingredients: 30 parts fish meal, 20 parts soybean meal, 15 parts corn meal, 5 parts astaxanthin, 10 parts β-glucan, 3 parts composite bacterial agent, and 17 parts complex A. The daily feeding amount of feed I was 2%-3% of the weight of the female broodstock, and it was fed 2-3 times a day. Feed II comprises the following raw materials by weight: 25 parts of fish meal, 25 parts of squid meal, 15 parts of salina powder, 10 parts of astaxanthin, 5 parts of rosmarinic acid, 5 parts of compound bacterial agent, and 15 parts of compound B; the daily feeding amount of Feed II is 1%-2% of the weight of male broodstock, and it is fed 2-3 times a day; The composite bacterial agent is composed of Acetobacter xylinum, Bacillus pumilus and Bacillus natto in a mass ratio of 5:3.5:1.5; Compound A comprises the following raw materials in parts by weight: 30 parts of Dendrobium officinale, 25 parts of Gynostemma pentaphyllum, 20 parts of ramie root, 15 parts of Aralia dasyphylla leaves, and 10 parts of Perilla seeds; Compound B comprises the following raw materials in parts by weight: 25 parts of seahorse, 25 parts of Cistanche deserticola, 20 parts of Acanthopanax senticosus, 15 parts of water chestnut shell, and 15 parts of Moringa oleifera seeds; During the parent cultivation process, 24 hours before the rainstorm, the enhancer was added at a rate of 0.5g / m 2 The enhancer includes the following raw materials in parts by weight: 30 parts of Aralia dasyphylla extract, 5 parts of L-valine, 10 parts of thiamine, 15 parts of disodium edetate, 20 parts of magnesium sulfate, and 20 parts of calcium dihydrogen phosphate.

[0025] (3) Fertilization and hatching: When the parent fish reach sexual maturity and spawn naturally, the fertilized eggs are transferred to an incubator (with seawater) for hatching. During the hatching process, the water temperature is controlled at 22-28°C, the salinity is 28‰~30‰, the pH value is 7.5~8.5, and the oxygen concentration is 4-6 mg / L. Before adding the fertilized eggs, 0.08% of the seawater mass of pullulan is added and plasma treated at 65W and 45kHz for 3 minutes. The fertilized eggs are added (fertilized egg density is 30,000-40,000 / m 3 ), and then magnetized at 65Gs for 25min, and the fry were obtained after hatching. Example 2 An artificial breeding method for hybrid pine tiger spot, comprising the following steps: (1) Parent selection: Select healthy freshwater groupers aged 3-4 years as female parents; select healthy brown-spotted groupers aged 1-2 years as male parents.

[0026] (2) Parent cultivation: Cultivation environment: Choose an open-air high-level pond with a water depth of 2-4m for cultivation, with a cultivation density of 3-5 fish / m 3 ; Three months before breeding, female broodstock were fed fortified feed I, and male broodstock were fed fortified feed II. Feed I included the following ingredients: 25 parts fish meal, 22 parts soybean meal, 20 parts corn meal, 3 parts astaxanthin, 8 parts β-glucan, 2 parts compound bacterial agent, and 20 parts compound A. The daily feeding amount of feed I was 2%-3% of the weight of the broodstock, and it was fed 2-3 times a day. Feed II includes the following raw materials in parts by weight: 20 parts of fish meal, 30 parts of squid meal, 10 parts of salina powder, 12 parts of astaxanthin, 5 parts of rosmarinic acid, 5 parts of compound bacterial agent, and 15 parts of compound B; the daily feeding amount of Feed II is 1%-2% of the weight of the parent, and it is fed 2-3 times a day; The composite bacterial agent is composed of Acetobacter xylinum, Bacillus pumilus and Bacillus natto in a mass ratio of 6:3:1; Compound A comprises the following raw materials in parts by weight: 27 parts of Dendrobium officinale, 30 parts of Gynostemma pentaphyllum, 15 parts of ramie root, 20 parts of Aralia dasyphylla leaves, and 8 parts of Perilla seeds; Compound B comprises the following raw materials in parts by weight: 23 parts of seahorse, 30 parts of Cistanche deserticola, 17 parts of Acanthopanax senticosus, 10 parts of water chestnut shell, and 20 parts of Moringa oleifera seeds; During the parent cultivation process, 24 hours before the rainstorm, the enhancer was added at a rate of 0.5g / m 2 The enhancer includes the following raw materials in parts by weight: 28 parts of Aralia dasyphylla extract, 4 parts of L-valine, 8 parts of thiamine, 20 parts of disodium edetate, 15 parts of magnesium sulfate, and 25 parts of calcium dihydrogen phosphate.

[0027] (3) Fertilization and hatching: When the parent fish reach sexual maturity and spawn naturally, the fertilized eggs are transferred to an incubator (with seawater) for hatching. During the hatching process, the water temperature is controlled at 22-28°C, the salinity is 28‰~30‰, the pH value is 7.5~8.5, and the oxygen concentration is 4-6 mg / L. Before adding the fertilized eggs, 0.05% of the seawater mass of pullulan is added and plasma treated at 60W and 40kHz for 4 minutes. The fertilized eggs are added (fertilized egg density is 30,000-40,000 particles / m 3 ), and then magnetized at 50Gs for 30min, and the fry were obtained after hatching. Example 3 An artificial breeding method for hybrid pine tiger spot, comprising the following steps: (1) Parent selection: Select healthy freshwater groupers aged 3-4 years as female parents; select healthy brown-spotted groupers aged 1-2 years as male parents.

[0028] (2) Parent cultivation: Cultivation environment: Choose an open-air high-level pond with a water depth of 2-4m for cultivation, with a cultivation density of 3-5 fish / m 3 ; Three months before breeding, female broodstock were fed fortified feed I, and male broodstock were fed fortified feed II. Feed I included the following ingredients: 35 parts of fish meal, 15 parts of soybean meal, 10 parts of corn meal, 8 parts of astaxanthin, 12 parts of β-glucan, 4 parts of compound bacterial agent, and 16 parts of compound A. The daily feeding amount of feed I was 2%-3% of the weight of the broodstock, and it was fed 2-3 times a day. Feed II includes the following raw materials in parts by weight: 30 parts of fish meal, 20 parts of squid meal, 18 parts of salina powder, 8 parts of astaxanthin, 3 parts of rosmarinic acid, 8 parts of compound bacterial agent, and 13 parts of compound B; the daily feeding amount of Feed II is 1%-2% of the weight of the parent, and it is fed 2-3 times a day; The composite bacterial agent is composed of Acetobacter xylinum, Bacillus pumilus and Bacillus natto in a mass ratio of 4:4:2; Compound A comprises the following raw materials in parts by weight: 33 parts of Dendrobium officinale, 20 parts of Gynostemma pentaphyllum, 25 parts of ramie root, 10 parts of Aralia dasyphylla leaves, and 12 parts of Perilla seeds; Compound B comprises the following raw materials in parts by weight: 28 parts of seahorse, 20 parts of Cistanche deserticola, 22 parts of Acanthopanax senticosus, 20 parts of water chestnut shell, and 10 parts of Moringa oleifera seeds; During the parent cultivation process, 24 hours before the rainstorm, the enhancer was added at a rate of 0.5g / m 2 The enhancer includes the following raw materials in parts by weight: 32 parts of Aralia dasyphylla extract, 6 parts of L-valine, 12 parts of thiamine, 10 parts of disodium edetate, 25 parts of magnesium sulfate, and 15 parts of calcium dihydrogen phosphate.

[0029] (3) Fertilization and hatching: When the parent fish reach sexual maturity and spawn naturally, the fertilized eggs are transferred to an incubator (with seawater) for hatching. During the hatching process, the water temperature is controlled at 22-28°C, the salinity is 28‰~30‰, the pH value is 7.5~8.5, and the oxygen concentration is 4-6 mg / L. Before adding the fertilized eggs, 0.1% of the seawater mass of pullulan is added and plasma treated at 70W and 50kHz for 2 minutes. The fertilized eggs are added (fertilized egg density is 30,000-40,000 / m 3 ), and then magnetized at 80Gs for 20min, and the fry were obtained after hatching. Comparative Example 1 The main difference from Example 1 is that the compound A does not contain ramie root and Aralia dasyphylla leaf, and the amount of Dendrobium officinale and Gynostemma pentaphyllum is increased in equal amounts. The compound A comprises the following raw materials in parts by weight: 50 parts of Dendrobium officinale, 40 parts of Gynostemma pentaphyllum, and 10 parts of Perilla seeds.

[0030] Comparative Example 2 The main difference from Example 1 is that Compound B does not contain Acanthopanax senticosus and Water Chestnut Shell, and the amount of Seahorse and Cistanche deserticola is increased in equal amounts. Compound B comprises the following raw materials in parts by weight: 45 parts of Seahorse, 40 parts of Cistanche deserticola, and 15 parts of Moringa seeds; Comparative Example 3 The main difference from Example 1 is that Acetobacter xylinum in the composite bacterial agent is replaced by Lactobacillus rhamnosus, wherein the composite bacterial agent is composed of Lactobacillus rhamnosus, Bacillus pumilus and Bacillus natto in a mass ratio of 5:3.5:1.5; Comparative Example 4 The main difference from Example 1 is that the Aralia dahurica extract is replaced by the Gentiana scabra root extract.

[0031] During the parent cultivation process, 24 hours before the rainstorm, the enhancer was added at a rate of 0.5g / m 2 The enhancer includes the following raw materials in parts by weight: 30 parts of gentian root extract, 5 parts of L-valine, 10 parts of thiamine, 15 parts of disodium edetate, 20 parts of magnesium sulfate, and 20 parts of calcium dihydrogen phosphate.

[0032] Test example The breeding methods of Examples 1-3 and Comparative Examples 1-4 were used to conduct experiments, with 200 female broodstock in each group. The egg production, fertilization rate, hatching rate, fry deformity rate, etc. were statistically analyzed, and the average values ​​were calculated, as shown in Table 1 below.

[0033] Table 1 - Artificial breeding of hybrid tiger spotted pine

[0034] As can be seen from the table above, the single female fish of Example 1 has the highest egg production, the highest fertilization rate and hatching rate, and no fry deformities. Examples 2-3 also have a high single female fish production, high fertilization rate and hatching rate, and low fry deformity rate.

[0035] Compared to Example 1, in Comparative Example 1, Composite A lacked ramie root and Aralia oleracea leaves, but instead incorporated equal amounts of Dendrobium officinale and Gynostemma pentaphyllum. Comparative Example 1 showed significant decreases in egg production, fertilization rate, and hatchability, and an increase in larval deformity rates. Composite A, prepared using ramie root and Aralia oleracea leaves in combination with Dendrobium officinale, Gynostemma pentaphyllum, and Perilla seed, promotes gonadal maturation in female fish, increases both egg production and egg quality, improves fertilization and hatchability, and reduces deformity rates.

[0036] In Comparative Example 2, Compound B did not contain Acanthopanax senticosus or Water Caltrop shells, but instead increased the amount of Hippocampus and Cistanche deserticola in equal amounts. The egg production, fertilization rate, and hatching rate of Comparative Example 2 all decreased significantly, and the larval deformity rate increased significantly. Compound B, prepared using Acanthopanax senticosus and Water Caltrop shells in combination with Hippocampus, Cistanche deserticola, and Moringa seeds, promotes gonadal maturation in male fish, improves semen quality, significantly increases fertilization rate and hatching rate, and significantly reduces deformity rate.

[0037] In Comparative Example 3, the composite bacterial agent, Acetobacter xylinum, was replaced with Lactobacillus rhamnosus. The prepared feeds I and II had an impact on the gonadal development of both male and female seven-headed starfish, affecting fertilization, resulting in a decrease in fertilized egg quality, a lower hatchability, and an increase in fry deformity rate. The present invention uses Acetobacter xylinum, Bacillus pumilus, and Bacillus natto to prepare feeds I and II, which are more conducive to improving the gonadal development of male and female fish and improving the quality of fertilized eggs.

[0038] In comparative example 4, the Aralia dahurica extract was replaced with the gentian root extract, and the breeding effect was significantly reduced. The enhancer of the present invention uses the gentian root extract in combination with L-valine, thiamine, disodium ethylenediaminetetraacetic acid, magnesium sulfate, and calcium dihydrogen phosphate, which is beneficial for reducing the adverse effects of heavy rain on the parents.

[0039] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the scope of the present invention should be included in the scope of protection of the present invention.

Claims

1. An artificial breeding method for hybrid spotted pine tiger, characterized in that: The following steps are involved: (1) Parent selection: Select healthy 3-4-year-old freshwater grouper as female parent; select healthy 1-2-year-old brown-spotted grouper as male parent; (2) Parent cultivation: 3 months before breeding, the female broodstock were fed with fortified feed I and the male broodstock were fed with fortified feed II; the feed I included the following raw materials: fish meal, soybean meal, corn meal, astaxanthin, β-glucan, compound bacteria agent, and compound A; the feed II included the following raw materials: fish meal, squid meal, salt algae meal, astaxanthin, rosmarinic acid, compound bacteria agent, and compound B; the compound bacteria agent included the following raw materials: Acetobacter xylinum, Bacillus pumilus, and Bacillus natto; the compound A included the following raw materials: Dendrobium officinale, Gynostemma pentaphyllum, ramie root, Aralia oleifera leaf, and Perilla seed; the compound B included the following raw materials: seahorse, Cistanche deserticola, Acanthopanax senticosus, water chestnut shell, and Moringa seed; (3) Fertilization and hatching: When the parent fish reach sexual maturity, they will naturally lay eggs and fertilize them. The fertilized eggs will be transferred to an incubator for hatching to obtain fry.

2. The artificial breeding method of the hybrid spotted spruce according to claim 1, characterized in that: The environment for parent cultivation: select an open-air high-level pond with a water depth of 2-4m for cultivation, and a cultivation density of 3-5 tails / m 3 .

3. The artificial breeding method of the hybrid spotted spruce according to claim 1, characterized in that: The feed I comprises the following raw materials in parts by weight: 25-35 parts of fish meal, 15-25 parts of soybean meal, 10-20 parts of corn flour, 3-8 parts of astaxanthin, 8-12 parts of β-glucan, 2-4 parts of composite bacterial agent, and 15-20 parts of complex A; the daily feeding amount of the feed I is 2%-3% of the weight of the parent, and it is fed 2-3 times a day.

4. The artificial breeding method of the hybrid spotted spruce according to claim 1, characterized in that: The feed II comprises the following raw materials in parts by weight: 20-30 parts of fish meal, 20-30 parts of squid meal, 10-20 parts of salt algae powder, 8-12 parts of astaxanthin, 3-8 parts of rosmarinic acid, 3-8 parts of compound bacterial agent, and 13-18 parts of complex B; the daily feeding amount of the feed II is 1%-2% of the weight of the parent, and it is fed 2-3 times a day.

5. The artificial breeding method of the hybrid spotted pine tiger according to claim 1 or 3, characterized in that: The compound A comprises the following raw materials in parts by weight: 25-35 parts of Dendrobium officinale, 20-30 parts of Gynostemma pentaphyllum, 15-25 parts of ramie root, 10-20 parts of Aralia dasyphylla leaves, and 8-12 parts of Perilla seeds.

6. The artificial breeding method of the hybrid spotted pine tiger according to claim 1 or 4, characterized in that: The compound B comprises the following raw materials in parts by weight: 20-30 parts of seahorse, 20-30 parts of Cistanche deserticola, 15-25 parts of Acanthopanax senticosus, 10-20 parts of water chestnut shells, and 10-20 parts of Moringa seeds.

7. The artificial breeding method of the hybrid spotted pine tiger according to claim 1, characterized in that: The mass ratio of Acetobacter xylinum, Bacillus pumilus and Bacillus natto in the composite bacterial agent is 4-6:3-4:1-2.

8. The artificial breeding method of the hybrid spotted pine tiger according to claim 1, characterized in that: During the parent cultivation process, 24 hours before the rainstorm, increase the dosage of the enhancer to 0.4-0.6g / m 2 The enhancer includes the following raw materials in parts by weight: 25-35 parts of Aralia dasyphylla extract, 4-6 parts of L-valine, 8-12 parts of thiamine, 10-20 parts of disodium ethylenediaminetetraacetic acid, 15-25 parts of magnesium sulfate, and 15-25 parts of calcium dihydrogen phosphate.

9. The artificial breeding method of the hybrid spotted pine tiger according to claim 1, characterized in that: Before adding fertilized eggs, seawater is placed in the incubator, pullulan is added, and plasma treatment is performed at 60-70W and 40-50kHz for 2-4 minutes. The amount of pullulan added is 0.05%-0.1% of the seawater.

10. The artificial breeding method of the hybrid spotted pine tiger according to claim 1 or 9, characterized in that: After adding the fertilized eggs, magnetization treatment was performed at 50-80 Gs for 20-30 minutes.

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