A medium-sized shrimp hard shell accelerator and its production method

Abstract

The invention discloses a medium-sized shrimp hard shell promoter and a production method thereof, and belongs to the technical field of aquatic feed additives. The medium-sized shrimp hard shell promoter of the invention comprises a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2-3 mL:100 g; the composite additive comprises the following components by mass percentage: 2.5% gentian, 2.5% orientalis, 3.5% ajuga, 10.0% calcium lactate, 2.0% calcium gluconate, 20.0% calcium dihydrogen phosphate, 1.0% copper glycinate, 0.5% glycinate iron, 4.0% magnesium sulfate, 1.0% zinc sulfate, 2.2% vitamin C, 3.8% vitamin D and 47% carrier zeolite powder. The medium-sized shrimp hard shell promoter of the invention can promote shrimp molting and hard shelling, and significantly improve the growth rate and survival rate of shrimp.

Description

Technical Field

[0001] The invention relates to the technical field of aquatic feed additives, in particular to a medium-sized shrimp hard shell promoter and a production method thereof. Background Art

[0002] The white shrimp is a marine invertebrate decapod Penaeididae, also known as white shrimp or white shrimp. The hard shell of the white shrimp can protect its body, but it also needs to molt periodically to achieve the purpose of growth.

[0003] The growth rate of white shrimp is affected by the molting rate and growth value-added rate (weight gain from each molting to the next molting), and the molting rate is related to water quality environmental factors, the physical strength of the shrimp, pathogens, enemies and even the invasion of the same species. The growth rate of mid-adult shrimp is the main factor determining the economic benefits of the shrimp farming industry. With the reduction of trace elements in the aquaculture water, the fluctuation of feed raw material quality, and the degradation of seedlings, the normal molting and hard shell condition of mid-adult shrimp plays an increasingly important role in the healthy growth of the shrimp body.

[0004] At present, there have been preliminary studies on the molting and hardening of shells of white shrimp in China, but there are few studies on promoting molting and hardening of shells in the middle-aged shrimp stage. After research, it was found that a nutritionally comprehensive shrimp feed can promote molting, but there are relatively few studies on compound feed premix additives that promote hardening and molting of middle-aged shrimp. How to effectively promote molting and hardening of shells in middle-aged shrimp has become a major problem in the healthy and efficient breeding of this variety. Summary of the invention

[0005] The purpose of the present invention is to provide a medium-sized shrimp hard shell promoter and a production method thereof to solve the problems existing in the above-mentioned prior art. The purpose of the present invention is to provide a medium-sized shrimp hard shell promoter that effectively promotes shrimp molting and hard shelling in order to solve the problems of difficult molting, slow hard shelling, decreased growth rate and survival rate during shrimp farming.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] One of the technical solutions of the present invention is: a medium-sized shrimp hard shell promoter, the components of which include a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2-3 mL: 100 g;

[0008] The composite additive comprises the following components, measured by mass percentage: 2.5% gentian, 2.5% oriental rhizome, 3.5% ajuga, 10.0% calcium lactate, 2.0% calcium gluconate, 20.0% calcium dihydrogen phosphate, 1.0% copper glycinate, 0.5% iron glycinate, 4.0% magnesium sulfate, 1.0% zinc sulfate, 2.2% vitamin C, 3.8% vitamin D and 47% carrier zeolite powder.

[0009] Furthermore, the method for preparing the Bacillus subtilis metabolites comprises the following steps:

[0010] inoculating the activated Bacillus subtilis into a seed culture medium, and obtaining a seed liquid after culturing;

[0011] inoculating the seed liquid into a fermentation medium for aerobic fermentation to obtain a fermentation culture liquid;

[0012] The fermentation culture liquid is centrifuged to obtain the supernatant, which is the Bacillus subtilis metabolite.

[0013] Furthermore, the components of the seed culture medium are: 10 g / L peptone, 5 g / L yeast powder, 5 g / L glucose and 10 g / L sodium chloride; the pH of the seed culture medium is 7.3-7.5;

[0014] The culture temperature when preparing the seed solution is 36-38°C and the time is 18-24 hours;

[0015] The components of the fermentation medium are: 20.0 g / L sucrose, 1.0 g / L sodium nitrate, 2.0 g / L potassium dihydrogen phosphate, 2.0 g / L disodium hydrogen phosphate, 1.0 g / L yeast powder and 1.0 g / L D-2,3,5,6-tetrahydroxy-2-hexenoic acid-γ-lactone; the pH of the fermentation medium is 7.2-7.5;

[0016] The temperature of the aerobic fermentation culture is 30-35° C. and the time is 48-72 hours.

[0017] Gentian, Alisma, and Ajuga have the function of promoting the body's metabolism, and have liver protection, anti-inflammatory, antibacterial, and antiviral effects. They can effectively improve the shrimp's resistance to adversity, improve the shrimp's physical fitness and survival rate, and enhance the shrimp's molting ability.

[0018] Combining organic and inorganic calcium sources (calcium lactate, calcium gluconate, and calcium dihydrogen phosphate) can effectively supplement the calcium deficiency of middle-aged shrimp when their shells harden after molting.

[0019] Combining organic and inorganic trace elements (glycinate copper, glycinate iron, magnesium sulfate, zinc sulfate) can regulate various stress factors in the molting and hardening of the shell of medium-sized shrimp to the greatest extent, and promote the molting and hardening of the shell of medium-sized shrimp. Vitamin C can significantly improve the vitality of the body, and vitamin D can promote the absorption of trace elements, which have a good promoting effect on the hardening of the shell of medium-sized shrimp.

[0020] The metabolites of Bacillus subtilis are rich in lipopeptides, which can stimulate the molting of shrimp, increase the molting speed and rate of shrimp, and thus improve the survival rate of shrimp.

[0021] Furthermore, the components of the medium-sized shrimp hard shell promoter also include probiotic particles and / or composite bacterial agents;

[0022] The dosage of the probiotic granules is 1-2% of the mass of the composite additive; the dosage of the composite bacterial agent is 0.8-1.2% of the mass of the composite additive.

[0023] Furthermore, the composite bacterial agent comprises the following components in parts by weight: 4 to 5 parts of Bacillus gelatinosarum, 2 to 3 parts of Bacillus mesosporus and 1 to 2 parts of Achromobacter xylosoxidans.

[0024] Furthermore, the probiotic granules include the following raw materials in parts by weight: 5 to 10 parts of bran, 10 to 15 parts of corn flour, 3 to 5 parts of lactose, 20 to 30 parts of corn stalk powder, 2 to 3 parts of multivitamins and 1 to 1.5 parts of probiotic powder;

[0025] The probiotic powder comprises the following components in parts by weight: 2-3 parts of Bacillus subtilis, 2-3 parts of Clostridium butyricum, 3-4 parts of Bacillus licheniformis and 2-3 parts of Saccharomyces cerevisiae.

[0026] Furthermore, the preparation method of the probiotic granules comprises the following steps:

[0027] The bran, corn flour, lactose, corn stalk powder and compound vitamins are mixed and crushed, and then the probiotic powder is added, mixed evenly and fermented to obtain a fermented product;

[0028] The starch solution is sprayed on the fermented product for granulation to obtain the probiotic granules.

[0029] Furthermore, the fermentation temperature is 20-30°C and the fermentation time is 8-10 days;

[0030] The particle size of the granulation is 1.5 to 2.0 mm.

[0031] The gelatinous Bacillus, Brevibacillus centosporus and Achromobacter xylosoxidans in the composite bacterial agent can degrade calcium phosphate that is difficult to be absorbed by shrimp into absorbable phosphorus and calcium, increase the phosphorus and calcium content in the shrimp, and promote the hardening of the shrimp shell.

[0032] Probiotic granules can improve the intestinal health of shrimp, promote the shrimp's absorption of nutrients, enhance the shrimp's ability to resist external bacteria during molting, prevent shrimp from being infected with harmful bacteria, and increase the shrimp's survival rate and growth rate.

[0033] The second technical solution of the present invention is a method for producing the above-mentioned medium-sized shrimp hard shell promoter, comprising the following steps:

[0034] The component raw materials are mixed evenly to obtain the medium-sized shrimp hard shell promoter.

[0035] The third technical solution of the present invention is to use the above-mentioned medium-sized shrimp hard shell promoter in promoting the molting and hard shelling of medium-sized shrimp.

[0036] Furthermore, the application method specifically includes: adding the medium-aged shrimp hard shell promoter to shrimp feed, and then feeding the shrimp; the addition amount of the medium-aged shrimp hard shell promoter in the shrimp feed is 0.1-1wt.%.

[0037] The present invention discloses the following technical effects:

[0038] (1) The hard shell promoter of the middle-aged shrimp of the present invention can promote the molting and hard shell formation of shrimps, and significantly improve the growth rate and survival rate of shrimps.

[0039] (2) The raw materials used in the medium-sized shrimp hard shell promoter of the present invention are easy to obtain, stable, low in price, simple in production process, free of side effects, non-toxic, and without hormone addition, and do not cause pollution to the environment, do not affect the normal life of aquatic animals, will not remain in the animal body, are non-corrosive, safe, hygienic, and quick to use, with low economic cost, and are suitable for large-scale use.

[0040] (3) The hard shell promoter of the middle-aged shrimp of the present invention can reduce the self-mutilation phenomenon of cultured shrimps and improve the survival rate; and can also improve the immunity of shrimps, improve the antibacterial and antiviral abilities, and improve the survival rate. DETAILED DESCRIPTION

[0041] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0042] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0043] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0044] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to the skilled artisan. The present invention description and examples are exemplary only.

[0045] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0046] The “parts” described in the following examples are all “parts by mass”.

[0047] The Bacillus subtilis used in the present invention is Bacillus subtilis YXSY-01, which is preserved in the Guangdong Provincial Microbiological Culture Collection Center, the preservation address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, the preservation number is: GDMCC No: 64078, and the preservation time is November 27, 2023.

[0048] The Clostridium butyricum, Bacillus licheniformis, Saccharomyces cerevisiae, Bacillus jelly, Bacillus centrosporus and Achromobacter xylosoxidans used in the present invention are all commercially available. The effective viable bacteria count of Clostridium butyricum is 2 billion / g, the effective viable bacteria count of Bacillus licheniformis is 500 million / g, the effective viable bacteria count of Saccharomyces cerevisiae is 5 billion / g, the effective viable bacteria count of Bacillus jelly is 5 billion / g, the effective viable bacteria count of Bacillus centrosporus is 200 million / g and the effective viable bacteria count of Achromobacter xylosoxidans is 300 million / g.

[0049] The composite vitamin adopted by the present invention consists of the following components: 2.0g of vitamin A, 2.5g of vitamin D3, 3.5g of vitamin E, 3.5g of vitamin K, 2.5g of vitamin B1, 3.0g of vitamin B2, 2.0g of vitamin B6 and 1.0g of vitamin B12.

[0050] The white shrimp feed used in the present invention is purchased from Jiangmen Xinhui District Puhui Aquatic Feed Co., Ltd., and the main ingredients of the feed are fish meal, soybean meal, high-gluten flour, peanut bran, yeast paste, chicken frame meat meal, calcium dihydrogen phosphate, soybean oil and lecithin oil.

[0051] Example 1

[0052] A production method of a medium-sized shrimp hard shell accelerator:

[0053] (1) a medium-sized shrimp hard shell promoter, comprising a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2.5 mL:100 g;

[0054] The composite additive is composed of the following components, measured by mass percentage: 2.5% of gentian, 2.5% of oriental water chestnut, 3.5% of ajuga, 10.0% of calcium lactate, 2.0% of calcium gluconate, 20.0% of calcium dihydrogen phosphate, 1.0% of copper glycinate, 0.5% of iron glycinate, 4.0% of magnesium sulfate, 1.0% of zinc sulfate, 2.2% of vitamin C, 3.8% of vitamin D and 47% of carrier zeolite powder.

[0055] Wherein, the preparation method of Bacillus subtilis metabolites is as follows:

[0056] A. streak Bacillus subtilis on the activated solid culture medium using an inoculation loop, and culture it in a constant temperature incubator at 37°C for 12 hours to obtain activated Bacillus subtilis;

[0057] Preparation of activated solid culture medium: Add 5.0 g of yeast powder, 10.0 g of peptone, 10.0 g of sodium chloride, and 20.0 g of agar to 500 mL of distilled water, stir evenly, add distilled water to make up to 1 L, adjust the pH to 7.2, and sterilize at 120°C for 20 min.

[0058] B. Pick a ring of activated Bacillus subtilis and inoculate it into 50 mL of seed culture medium. After culturing (temperature 36° C., time 24 h, rotation speed 200 rpm), obtain seed liquid.

[0059] Preparation of seed culture medium: Add 10 g / L peptone, 5 g / L yeast powder, 5 g / L glucose, and 10 g / L sodium chloride to 500 mL of distilled water, stir evenly, add distilled water to make up to 1 L, adjust the pH to 7.5, and sterilize at 120°C for 20 min.

[0060] C. The seed liquid was inoculated into the fermentation medium (the volume ratio of the seed liquid to the fermentation medium was 1:50), and then aerobic fermentation culture was carried out (temperature was 32°C, time was 72h, rotation speed was 200rpm) to obtain the fermentation culture liquid.

[0061] Preparation of fermentation medium: Add 20.0 g sucrose, 1.0 g sodium nitrate, 2.0 g potassium dihydrogen phosphate, 2.0 g disodium hydrogen phosphate, 1.0 g yeast powder, and 1.0 g D-2,3,5,6-tetrahydroxy-2-hexenoic acid-γ-lactone to 500 mL of distilled water, stir evenly, add distilled water to make up to 1 L, adjust the pH to 7.2, and sterilize at 120°C for 20 min.

[0062] D. The supernatant obtained after centrifuging the fermentation culture broth (at 8000 rpm for 10 min) is the metabolite of Bacillus subtilis.

[0063] (2) Production method: The component raw materials (particle size greater than 30 mesh) are crushed by an ultra-fine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a medium-sized shrimp hard shell promoter.

[0064] Example 2

[0065] A production method of a medium-sized shrimp hard shell accelerator:

[0066] (1) a medium-sized shrimp hard shell promoter, comprising a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2 mL:100 g;

[0067] The composite additive is composed of the following components, measured by mass percentage: 2.5% of gentian, 2.5% of oriental water chestnut, 3.5% of ajuga, 10.0% of calcium lactate, 2.0% of calcium gluconate, 20.0% of calcium dihydrogen phosphate, 1.0% of copper glycinate, 0.5% of iron glycinate, 4.0% of magnesium sulfate, 1.0% of zinc sulfate, 2.2% of vitamin C, 3.8% of vitamin D and 47% of carrier zeolite powder.

[0068] Wherein, the preparation method of Bacillus subtilis metabolites is as follows:

[0069] A. streak Bacillus subtilis on the activated solid culture medium using an inoculation loop, and culture it in a constant temperature incubator at 37°C for 12 hours to obtain activated Bacillus subtilis;

[0070] Preparation of activated solid culture medium: Add 5.0 g of yeast powder, 10.0 g of peptone, 10.0 g of sodium chloride, and 20.0 g of agar to 500 mL of distilled water, stir evenly, add distilled water to make up to 1 L, adjust the pH to 7.2, and sterilize at 120°C for 20 min.

[0071] B. Pick a ring of activated Bacillus subtilis and inoculate it into 50 mL of seed culture medium. After culturing (temperature 38° C., time 18 h, rotation speed 200 rpm), obtain seed liquid.

[0072] Preparation of seed culture medium: Add 10 g / L peptone, 5 g / L yeast powder, 5 g / L glucose, and 10 g / L sodium chloride to 500 mL of distilled water, stir evenly, add distilled water to make up to 1 L, adjust the pH to 7.3, and sterilize at 120°C for 20 min.

[0073] C. The seed liquid was inoculated into the fermentation medium (the volume ratio of the seed liquid to the fermentation medium was 1:50), and then aerobic fermentation culture was carried out (temperature was 30°C, time was 48h, rotation speed was 200rpm) to obtain the fermentation culture liquid.

[0074] Preparation of fermentation medium: Add 20.0 g sucrose, 1.0 g sodium nitrate, 2.0 g potassium dihydrogen phosphate, 2.0 g disodium hydrogen phosphate, 1.0 g yeast powder, and 1.0 g D-2,3,5,6-tetrahydroxy-2-hexenoic acid-γ-lactone to 500 mL of distilled water, stir evenly, add distilled water to make the volume to 1 L, adjust the pH to 7.5, and sterilize at 120°C for 20 min.

[0075] D. The supernatant obtained after centrifuging the fermentation culture broth (at 8000 rpm for 10 min) is the metabolite of Bacillus subtilis.

[0076] (2) Production method: The component raw materials (particle size greater than 30 mesh) are crushed by an ultra-fine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a medium-sized shrimp hard shell promoter.

[0077] Comparative Example 1

[0078] Same as Example 1, except that Ajuga acuminata is replaced with Atractylodes macrocephala in an equal mass percentage.

[0079] Comparative Example 2

[0080] Same as Example 1, except that Alisma orientalis and Ajuga strychnifolia are replaced by Gentiana scabra in percentage by mass.

[0081] Comparative Example 3

[0082] The same as Example 1, except that calcium gluconate and calcium dihydrogen phosphate are replaced by calcium lactate in equal mass percentages.

[0083] Comparative Example 4

[0084] The same as Example 1, except that copper glycinate is replaced by copper sulfate in an equal mass percentage; and iron glycinate is replaced by ferrous sulfate in an equal mass percentage.

[0085] Example 3

[0086] A production method of a medium-sized shrimp hard shell promoter (adding probiotic particles):

[0087] (1) The medium-sized shrimp hard shell promoter is composed of probiotic particles, Bacillus subtilis metabolites and composite additives in a dosage ratio of 2.5 mL: 100 g;

[0088] The amount of the probiotic granules used is 1.5% of the mass of the composite additive.

[0089] The preparation method of the Bacillus subtilis metabolite and the ingredients of the composite additive are the same as those in Example 1.

[0090] Preparation of probiotic granules:

[0091] A. Mix and grind 8 parts of bran, 12 parts of corn flour, 5 parts of lactose, 25 parts of corn stalk powder and 3 parts of multivitamins, add 1.2 parts of probiotic powder, mix well and ferment (fermentation temperature is 25° C., time is 10 days) to obtain a fermentation product;

[0092] Among them, the probiotic powder is composed of the following components in parts by mass: 2.5 parts of Bacillus subtilis, 3 parts of Clostridium butyricum, 4 parts of Bacillus licheniformis and 2 parts of Saccharomyces cerevisiae.

[0093] B. The fermented product was dried and crushed to 200 mesh, and then sprayed with a 2 wt.% starch aqueous solution to form granules (particle size of 1.5 mm) to obtain probiotic granules.

[0094] (2) Production method: The component raw materials (particle size greater than 30 mesh) other than the probiotic granules are crushed by an ultrafine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a mixture.

[0095] The mixed material and the probiotic granules constitute the medium-sized shrimp hard shell promoter.

[0096] Example 4

[0097] A production method of a medium-sized shrimp hard shell promoter (adding a composite bacterial agent):

[0098] (1) The medium-sized shrimp hard shell promoter is composed of a composite bacterial agent, a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2.5 mL: 100 g;

[0099] The dosage of the composite bacterial agent is 1.2% of the mass of the composite additive.

[0100] The composite bacterial agent is composed of the following components in parts by mass: 4.5 parts of jelly-like spore bacillus, 2.5 parts of Brevibacillus mesosporus and 1.5 parts of Achromobacter xylosoxidans.

[0101] The preparation method of the Bacillus subtilis metabolite and the ingredients of the composite additive are the same as those in Example 1.

[0102] (2) Production method: The component raw materials (particle size greater than 30 mesh) are crushed by an ultra-fine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a medium-sized shrimp hard shell promoter.

[0103] Example 5

[0104] A method for producing a medium-sized shrimp hard shell promoter (adding probiotic granules and composite bacterial agents):

[0105] (1) The medium-sized shrimp hard shell promoter is composed of probiotic granules, composite bacterial agent, Bacillus subtilis metabolites and composite additives in a dosage ratio of 2.5 mL: 100 g;

[0106] The amount of probiotic granules used is 1.5% of the mass of the composite additive; the amount of the composite bacterial agent used is 1.2% of the mass of the composite additive.

[0107] The composite bacterial agent is composed of the following components in parts by mass: 4.5 parts of jelly-like spore bacillus, 2.5 parts of Brevibacillus mesosporus and 1.5 parts of Achromobacter xylosoxidans.

[0108] The preparation method of the Bacillus subtilis metabolite and the ingredients of the composite additive are the same as those in Example 1.

[0109] Preparation of probiotic granules:

[0110] A. Mix and grind 8 parts of bran, 12 parts of corn flour, 5 parts of lactose, 25 parts of corn stalk powder and 3 parts of multivitamins, add 1.2 parts of probiotic powder, mix well and ferment (fermentation temperature is 25° C., time is 10 days) to obtain a fermentation product;

[0111] Among them, the probiotic powder is composed of the following components in parts by mass: 2.5 parts of Bacillus subtilis, 3 parts of Clostridium butyricum, 4 parts of Bacillus licheniformis and 2 parts of Saccharomyces cerevisiae.

[0112] B. The fermented product was dried and crushed to 200 mesh, and then sprayed with a 2 wt.% starch aqueous solution to form granules (particle size of 1.5 mm) to obtain probiotic granules.

[0113] (2) Production method: The component raw materials (particle size greater than 30 mesh) other than the probiotic granules are crushed by an ultrafine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a mixture.

[0114] The mixed material and the probiotic granules constitute the medium-sized shrimp hard shell promoter.

[0115] Example 6

[0116] A method for producing a medium-sized shrimp hard shell promoter (adding probiotic granules and composite bacterial agents):

[0117] (1) The medium-sized shrimp hard shell promoter is composed of probiotic granules, composite bacterial agent, Bacillus subtilis metabolites and composite additives in a dosage ratio of 2.5 mL: 100 g;

[0118] The amount of probiotic granules used is 2% of the mass of the composite additive; the amount of the composite bacterial agent used is 0.8% of the mass of the composite additive.

[0119] The composite bacterial agent is composed of the following components in parts by mass: 5 parts of Bacillus gelatinosarum, 2 parts of Brevibacillus centosporus and 2 parts of Achromobacter xylosoxidans.

[0120] The preparation method of the Bacillus subtilis metabolite and the ingredients of the composite additive are the same as those in Example 1.

[0121] Preparation of probiotic granules:

[0122] A. Mix and grind 10 parts of bran, 10 parts of corn flour, 3 parts of lactose, 30 parts of corn stalk powder and 2 parts of multivitamins, add 1 part of probiotic powder, mix well and ferment (fermentation temperature is 30° C., time is 8 days) to obtain a fermented product;

[0123] Among them, the probiotic powder is composed of the following components in parts by mass: 3 parts of Bacillus subtilis, 2 parts of Clostridium butyricum, 3 parts of Bacillus licheniformis and 3 parts of Saccharomyces cerevisiae.

[0124] B. The fermented product was dried and crushed to 200 meshes, and then sprayed with a 2 wt.% starch aqueous solution to form granules (particle size of 2.0 mm) to obtain probiotic granules.

[0125] (2) Production method: The component raw materials (particle size greater than 30 mesh) other than the probiotic granules are crushed by an ultrafine grinder until they pass through a 30-mesh sieve, and then the component raw materials are mixed evenly to obtain a mixture.

[0126] The mixed material and the probiotic granules constitute the medium-sized shrimp hard shell promoter.

[0127] Comparative Example 5

[0128] Same as Example 3, except that the probiotic granules are replaced with a probiotic complex.

[0129] Preparation of probiotic complex: 8 parts of bran, 12 parts of corn flour, 5 parts of lactose, 25 parts of corn stalk powder, 3 parts of complex vitamins, and 1.2 parts of probiotic powder are crushed by ultrafine grinder respectively, passed through a 30-mesh sieve, and mixed evenly to obtain a probiotic complex.

[0130] The probiotic powder is composed of the following components in parts by mass: 2.5 parts of Bacillus subtilis, 3 parts of Clostridium butyricum, 4 parts of Bacillus licheniformis and 2 parts of Saccharomyces cerevisiae.

[0131] Effect Example 1

[0132] To determine the effects of hard shell promoters on the growth of white shrimp.

[0133] (1) Experimental site: Indoor aquaculture water system of Guangdong Nutriera Co., Ltd. (Zhuhai, Guangdong).

[0134] (2) Experimental time: 2023.

[0135] (3) Feed production method: The medium-sized shrimp hard shell promoter prepared in Example 1 is added to white shrimp feed (commercially available), mixed evenly, and a mixed feed is obtained.

[0136] The specific method is as follows:

[0137] Take clean tap water, and add 1g / kg (0.1wt.%), 2g / kg (0.2wt.%), 4g / kg (0.4wt.%), and 8g / kg (0.8wt.%) according to the amount of feed. Dissolve the hard shell promoter of the medium-sized shrimp prepared in Example 1 in tap water half an hour in advance (the amount of tap water is 5% of the mass of the white shrimp feed), stir evenly after dissolution, and then put them into a mixer together with the meal feed (the amount of feed fed per meal) and stir for 1 minute to allow the medium-sized shrimp hard shell promoter to fully penetrate into the feed, and then put them into a feed bag and leave them for 30 minutes for use.

[0138] (4) Experimental shrimp and culture system

[0139] The shrimp used in the experiment were 25-day-old whiteleg shrimp with a body length of 9±1cm and a body weight of 5±0.5g.

[0140] At the beginning of the experiment, 750 shrimps with strong physique and good vitality were selected and randomly divided into 5 groups, 150 shrimps in each group, 3 replicates (50 shrimps in each replicate), and stocked in 15 glass tanks (60×40×30cm, 72L). The experimental water was aerated tap water with adjusted salinity, with a water temperature of 25-27℃, a salinity of 5‰, and a pH of 7.8-8.6. Each glass tank was equipped with an air stone for aeration to keep the dissolved oxygen content above 6mg / L, and the amount of water change increased with the age. During the breeding experiment, the shrimps were fed at 6% of their body weight (twice a day, the total feeding amount was 6% of the shrimp body weight). The feces and residual bait at the bottom of the glass tank were sucked out with a bottom suction pump every day. Every other week, 5-6 white shrimps were randomly selected from each group of tanks to measure their weight, and the feeding amount was adjusted in time according to their weight. The experimental period was 56 days. The group fed only with white shrimp feed was used as the control group, and the rest were treated groups.

[0141] After one week of feeding, it was clearly observed that the shrimp shells in each treatment group were smoother, more transparent and harder than those in the control group, with fewer soft shells. During the experiment, the shrimps were healthy and ate normally. No obvious damage or adverse effects of the test feed on the shrimps were found.

[0142] The results of the effects of the hard shell promoter on the survival and growth of Penaeus vannamei are shown in Table 1.

[0143] Survival rate = (total number of shrimps - number of dead shrimps) / total number of shrimps × 100%;

[0144] Body weight gain (WG) = final body weight - initial body weight.

[0145] Table 1 Effects of shrimp hard shell promoter on survival and growth of Penaeus vannamei

[0146] Treatment Group Additive usage Survival rate (%) WG(g / tail) Control group 0wt.% 96.33±2.49 248.09±33.61 Treatment Group 1 0.1wt.% 97.67±0.94 272.76±39.22 Treatment Group 2 0.2wt.% 98.67±1.89 284.84±40.59 Treatment Group 3 0.4wt.% 98.00±1.63 270.76±24.37 Treatment Group 4 0.8wt.% 98.00±6.53 259.59±45.88

[0147] Note: The WG data in Table 1 are the mean ± standard deviation of 10 randomly selected shrimps.

[0148] As can be seen from Table 1, the medium-sized shrimp hard shell promoter prepared in Example 1 has a significant effect on the survival rate and WG of Penaeus vannamei. The survival rate and WG of the treatment group using the promoter prepared in Example 1 are significantly higher than those of the control group, and the appropriate addition amount of the medium-sized shrimp hard shell promoter is 0.2wt.%.

[0149] The test was conducted in four 1-mu earth ponds at the Guangdong Nutriera pilot base (Zhuhai, Guangdong) (January to April 2023), of which two earth ponds were fed with feed containing 0.2wt.% of the hard shell promoter for medium-sized shrimp (experimental pond 1 and experimental pond 2), and the other two earth ponds were fed with white shrimp feed without promoter (control pond 1 and control pond 2). After stocking (about 60,000 seedlings) and culturing to a size of 110 per catty, a 35-day feeding experiment with the addition of the hard shell promoter for medium-sized shrimp was conducted, and finally the data was collected after the fish were released from the pond.

[0150] Table 2 Effect of soil pond aquaculture

[0151] Pond No. Uniformity Specification Output Survival rate Control pond 1 Difference 32 pieces / catties 1100 catties 58.7％ Control pond 2 generally 38 pieces / catties 1197 catties 70.3％ Experimental Pond 1 good 32 pieces / catties 1486 catties 79.3％ Experimental Pond 2 good 35 pieces / catties 1250 catties 75.8％

[0152] As can be seen from Table 2, the experimental ponds fed with the hard shell promoter for mid-sized shrimps had significantly higher shrimp size uniformity, specifications, per mu yield (shrimp output) and survival rate than the control group. This indicates that the hard shell promoter for mid-sized shrimps has a good effect on mid-sized shrimps of Penaeus vannamei and can improve the economic benefits of white shrimps.

[0153] In summary, the medium-sized shrimp hard shell promoter prepared by the present invention can effectively promote the molting and hard shelling of white shrimp, improve the survival rate of shrimp in the breeding process, and has great practical value and economic benefits.

[0154] Effect Example 2

[0155] To determine the effects of hard shell promoters on the growth of Penaeus vannamei.

[0156] (1) Experimental site: Indoor aquaculture water system of Guangdong Nutriera Co., Ltd. (Zhuhai, Guangdong).

[0157] (2) Experimental time: 2023.

[0158] (3) Feed production method: The medium-sized shrimp hard shell promoter prepared in the embodiment and the comparative example is added to the white shrimp feed respectively, and mixed evenly to obtain a mixed feed.

[0159] The specific method is as follows:

[0160] A. Method of using the medium-sized shrimp hard shell accelerator prepared in Example 2 and Comparative Examples 1 to 4:

[0161] Take clean tap water, and add 2g / kg (0.2wt.%) according to the amount of feed, dissolve the hard shell promoter of medium-sized shrimp prepared in Example 2 and Comparative Examples 1 to 4 in tap water half an hour in advance (the amount of tap water is 5% of the mass of white shrimp feed), stir evenly after dissolution, and then put it into a blender together with the meal feed (the amount of feed fed per meal) and stir for 1min to allow the hard shell promoter of medium-sized shrimp to fully penetrate into the feed, and then put it into a feed bag and leave it for 30min for use.

[0162] B. Method for using the medium-sized shrimp hard shell accelerator prepared in Example 3:

[0163] Take clean tap water and add 2 g / kg (0.2 wt.%) according to the amount of feed. Dissolve the mixture prepared in Example 3 in tap water half an hour in advance (the amount of tap water is 5% of the mass of the white shrimp feed), stir evenly after dissolution, and then put it into a blender together with the probiotic granules and the meal feed (the amount of feed fed per meal) and stir for 1 minute to allow the mixture to fully penetrate into the feed, and then put it into a feed bag and leave it for 30 minutes for use.

[0164] C. The method of using the medium-sized shrimp hard shell promoter prepared in Example 4 is the same as step A.

[0165] D. The method of using the medium-sized shrimp hard shell promoter in Examples 5 and 6 is the same as step B.

[0166] E. The method of using the medium-sized shrimp hard shell promoter prepared in Comparative Example 5 is the same as that in step A.

[0167] (4) Experimental shrimp and culture system

[0168] The shrimp used in the experiment were 25-day-old whiteleg shrimp, 9±1cm in length, and 5±0.5g in weight.

[0169] At the beginning of the experiment, 1650 shrimps with strong physique and good vitality were selected and randomly divided into 11 groups, 150 shrimps in each group, divided into 3 replicates (50 shrimps in each replicate), and stocked in 33 glass tanks (60×40×30cm, 72L). The experimental water was aerated tap water with adjusted salinity, with a water temperature of 25-27℃, a salinity of 5‰, and a pH of 7.8-8.6. Each glass tank was equipped with an air stone for inflation to keep the dissolved oxygen content above 6mg / L, and the amount of water change increased with the age. During the breeding experiment, the shrimps were fed at 6% of their body weight (twice a day, the total feeding amount was 6% of the shrimp body weight). The feces and residual bait at the bottom of the glass tank were sucked out with a bottom suction pump every day. Every other week, 5-6 white shrimps were randomly selected from each group of tanks to measure their weight, and the feeding amount was adjusted in time according to their weight. The experimental period was 56 days. The group fed only with white shrimp feed was used as the control group, and the rest were treated groups.

[0170] The results of the effects of the hard shell promoter on the survival and growth of Penaeus vannamei are shown in Table 3.

[0171] Survival rate = (total number of shrimps - number of dead shrimps) / total number of shrimps × 100%;

[0172] Body weight gain (WG) = final body weight - initial body weight.

[0173] Table 3 Effects of shrimp hard shell promoter on survival and growth of Penaeus vannamei

[0174] Treatment Group Additive usage Survival rate (%) WG(g / tail) Control group 0wt.% 96.33±2.49 248.09±33.61 Example 2 0.2wt.% 98.67±0.94 285.40±9.41 Comparative Example 1 0.2wt.% 98.00±1.63 279.30±16.55 Comparative Example 2 0.2wt.% 97.33±0.94 273.60±20.80 Comparative Example 3 0.2wt.% 96.67±0.94 268.40±19.93 Comparative Example 4 0.2wt.% 96.67±2.49 274.20±21.77 Example 3 0.2wt.% 98.67±0.94 293.90±16.67 Example 4 0.2wt.% 98.00±1.63 290.10±12.91 Example 5 0.2wt.% 100±0.00 297.50±14.77 Example 6 0.2wt.% 99.33±0.94 296.10±11.62 Comparative Example 5 0.2wt.% 98.67±0.94 287.70±10.81

[0175] Note: The WG data in Table 3 are the mean ± standard deviation of 10 randomly selected shrimps.

[0176] The test was conducted in 6 1-mu earth ponds at the Guangdong Nutriera pilot base (Zhuhai, Guangdong) (January 2023 to April 2023), of which two earth ponds were fed with feeds (experimental ponds 1 and 2) with 0.2wt.% of the hard shell promoter of medium-sized shrimp (prepared in Example 5), two earth ponds were fed with feeds (experimental ponds 3 and 4) with 0.2wt.% of the hard shell promoter of medium-sized shrimp (prepared in Example 6), and two earth ponds were fed with white shrimp feeds without promoters (control ponds 1 and 2). After stocking (about 60,000 seedlings) and culturing to a specification of 110 tails / jin, a 35-day feeding experiment with the addition of the hard shell promoter of medium-sized shrimp was carried out, and finally the statistics were recorded when the fish were out of the pond.

[0177] Table 4 Effect of soil pond aquaculture

[0178] Pond No. Uniformity Specification Output Survival rate Experimental Pond 1 good 34 pieces / catties 1635 catties 88.4％ Experimental Pond 2 good 36 pieces / catties 1559 catties 87.2％ Experimental Pond 3 good 35 pieces / catties 1545 catties 84.7％ Experimental Pond 4 good 37 pieces / catties 1526 catties 86.2％ Control pond 1 Difference 33 pieces / catties 1059 catties 57.2％ Control pond 2 generally 34 pieces / catties 1125 catties 65.2％

[0179] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A medium-sized shrimp hard shell accelerator, characterized in that: The components include a Bacillus subtilis metabolite and a composite additive in a dosage ratio of 2 to 3 mL: 100 g; The composite additive comprises the following components by mass percentage: 2.5% gentian, 2.5% oriental rhizome, 3.5% ajuga, 10.0% calcium lactate, 2.0% calcium gluconate, 20.0% calcium dihydrogen phosphate, 1.0% copper glycinate, 0.5% iron glycinate, 4.0% magnesium sulfate, 1.0% zinc sulfate, 2.2% vitamin C, 3.8% vitamin D and 47% carrier zeolite powder; The components of the medium-sized shrimp hard shell promoter also include probiotic particles and composite bacterial agents; The amount of the probiotic granules is 1-2% of the mass of the composite additive; the amount of the composite bacterial agent is 0.8-1.2% of the mass of the composite additive; The composite bacterial agent comprises the following components in parts by weight: 4 to 5 parts of Bacillus jelly-like, 2 to 3 parts of Bacillus mesosporus and 1 to 2 parts of Achromobacter xylosoxidans; The probiotic granules include the following raw materials in parts by weight: 5 to 10 parts of bran, 10 to 15 parts of corn flour, 3 to 5 parts of lactose, 20 to 30 parts of corn stalk powder, 2 to 3 parts of complex vitamins, and 1 to 1.5 parts of probiotic powder; The probiotic powder comprises the following components in parts by weight: 2-3 parts of Bacillus subtilis, 2-3 parts of Clostridium butyricum, 3-4 parts of Bacillus licheniformis and 2-3 parts of Saccharomyces cerevisiae; The preparation method of the probiotic granules comprises the following steps: mixing and grinding bran, corn flour, lactose, corn stalk powder and complex vitamins, adding probiotic powder, mixing evenly and fermenting to obtain a fermented product; spraying a starch solution on the fermented product and granulating to obtain the probiotic granules; the fermentation temperature is 20-30° C. and the fermentation time is 8-10 days; the granulated particle size is 1.5-2.0 mm; The production method of the medium-sized shrimp hard shell promoter comprises the following steps: mixing component raw materials uniformly to obtain the medium-sized shrimp hard shell promoter.

2. The medium-sized shrimp hard shell accelerator according to claim 1, characterized in that The method for preparing the Bacillus subtilis metabolites comprises the following steps: inoculating the activated Bacillus subtilis into a seed culture medium, and obtaining a seed liquid after culturing; inoculating the seed liquid into a fermentation medium for aerobic fermentation to obtain a fermentation culture liquid; The fermentation culture liquid is centrifuged to obtain the supernatant, which is the Bacillus subtilis metabolite.

3. The medium-sized shrimp hard shell accelerator according to claim 2, characterized in that The components of the seed culture medium are: 10 g / L peptone, 5 g / L yeast powder, 5 g / L glucose and 10 g / L sodium chloride; the pH of the seed culture medium is 7.3-7.5; The culture temperature when preparing the seed solution is 36-38°C and the time is 18-24 hours; The components of the fermentation medium are: 20.0 g / L sucrose, 1.0 g / L sodium nitrate, 2.0 g / L potassium dihydrogen phosphate, 2.0 g / L disodium hydrogen phosphate, 1.0 g / L yeast powder and 1.0 g / L D-2,3,5,6-tetrahydroxy-2-hexenoic acid-γ-lactone; the pH of the fermentation medium is 7.2-7.5; The temperature of the aerobic fermentation culture is 30-35° C. and the time is 48-72 hours.

4. Use of the hard shell promoter of middle-aged shrimp according to any one of claims 1 to 3 in promoting molting and hard shelling of middle-aged shrimp.