CN116508912B

CN116508912B - A fishery regulating composition with lipid-lowering and blood sugar-controlling effects, and its preparation method and application

Info

Publication number: CN116508912B
Application number: CN202310633817.1A
Authority: CN
Inventors: 徐超; 沈慧超; 丁昊
Original assignee: South China Agricultural University
Current assignee: South China Agricultural University
Priority date: 2023-05-31
Filing date: 2023-05-31
Publication date: 2024-12-10
Anticipated expiration: 2043-05-31
Also published as: CN116508912A

Abstract

The present invention belongs to the technical field of aquaculture, and discloses a fishery regulating composition with lipid-lowering and blood sugar-controlling effects, and a preparation method and application thereof. The composition comprises the following components in parts by weight: 2 to 50 parts of berberine, 4 to 50 parts of allicin, 2 to 30 parts of benfotiamine, 2 to 50 parts of γ-aminobutyric acid, 4 to 20 parts of sodium oxalate, and 200 to 530 parts of γ-glutamyl-phenylalanine. Among them, γ-glutamyl-phenylalanine is obtained by fermenting a mixture of cotton stalks and corn stalks. The fishery regulating composition of the present invention makes full use of the different biological structures of the raw materials to carry out scientific compatibility and exert synergistic effects. By regulating the sugar and lipid metabolism function of farmed animals, the effect of lipid-lowering and blood sugar-control is achieved, thereby improving the sugar metabolism function of aquatic animals and significantly improving their utilization rate of carbohydrates in feed. The regulating composition is used in freshwater fish farming.

Description

Fishing regulation composition with lipid-lowering and blood sugar-controlling functions, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of aquaculture, and particularly relates to a fish regulation and control composition with the functions of reducing blood lipid and controlling blood sugar, and a preparation method and application thereof.

Background

The sugar is used as the cheapest energy source in the three nutrient substances, and the proper sugar level is added into the aquatic feed, so that the consumption of protein raw materials in the feed can be saved. However, fish are congenital diabetic constitutions, and have low sugar utilization, and persistent hyperglycemia symptoms occur after ingestion of high carbohydrate feeds. At the same time, fatty liver phenomena such as liver hypertrophy, liver color whitening, and increased cavitation of liver cells are accompanied by remarkably increased blood fat and liver lipid levels, and the phenomena can cause nutritional metabolic diseases, further cause the decrease of fish growth and even death. At present, few reports on development of aquatic animal sugar metabolism regulators are provided, and research on the sugar reducing effect of western medicines is mainly focused on. However, western medicines are easy to cause the problems of drug resistance, drug residue and the like of aquatic animals in the using process, so that the wide application of the western medicines in feeds is restricted. Meanwhile, the bioavailability of the aquatic animal sugar metabolism regulators sold in the market at present is low, which greatly limits the exertion of their functions.

In recent years, under the large trend of reducing resistance, replacing resistance and having no resistance in the national aquaculture industry, research on active ingredients of medicinal plants, high-efficiency vitamins and amino acid products is increasingly paid attention to. Berberine is also called berberine, which is a quaternary ammonium alkaloid in Chinese medicine coptis chinensis, allicin is an organic sulfur compound extracted from bulb (garlic bulb) of Allium plant of Alliaceae, benfotiamine is a derivative of thiamine (vitamin B1) and has higher bioavailability due to its fat solubility, gamma-aminobutyric acid is an amino acid composed of naturally occurring non-protein, is considered to be an important inhibitory neurotransmitter in the central nervous system of an organism, and sodium oxalate salt is an inhibitor of lactic dehydrogenase a in the glycolysis process of the organism. These substances have all been shown to regulate the body's sugar metabolism, but their mechanisms of action are markedly different. For example, gamma-aminobutyric acid enhances the sugar metabolism of brain cells by regulating the tricarboxylic acid cycle of brain tissue and increasing the activity of glucose phosphatase during glucose metabolism. Benfotiamine regulates the glucose metabolism of brain tissues by activating glycogen synthase kinase-3 mediated glucose metabolism signaling pathways. The oxalic acid sodium salt reduces the generation of endogenous lactic acid by inhibiting the activity of lactic acid dehydrogenase a, thereby regulating and controlling the sugar metabolism function of organism tissues. In view of the above, if the characteristics of different biological structures and action mechanisms of the 5 substances are utilized to carry out scientific compatibility and make the 5 substances play a synergistic role, then the composition for regulating and controlling the metabolism of the fish sugar with the functions of reducing blood lipid and controlling blood sugar is developed, and the composition has great significance in reducing the cost of feed and improving the economic benefit of cultivation.

The absorption and bioavailability of a regulatory composition directly affect its effectiveness, while increasing the phagostimulability of a regulatory composition is an important method of improving its absorption and bioavailability. The gamma-glutamyl-phenylalanine is a compound amino acid produced by microbial fermentation, can obviously improve the delicate flavor of foods (such as soy sauce and model chicken soup), and has strong phagostimulant property. In addition, the microencapsulation technology refers to a process of encapsulating bioactive substances (capsule cores) into tiny capsules by using natural or synthetic polymer materials (capsule materials), and can improve the stability and the bioavailability of the bioactive substances.

Disclosure of Invention

In order to solve the defects and shortcomings of the prior art, the invention aims to provide a fishing regulation composition with the functions of reducing blood lipid and controlling blood sugar.

The invention also aims at providing a preparation method of the fishing regulation composition with the functions of reducing blood lipid and controlling blood sugar.

It is still another object of the present invention to provide the use of the above-mentioned fishing control composition for lowering blood lipid and controlling blood glucose.

The aim of the invention is achieved by the following technical scheme:

The fishing regulation and control composition with the functions of reducing blood lipid and controlling blood sugar comprises, by weight, 2-50 parts of berberine, 4-50 parts of allicin, 2-30 parts of benfotiamine, 2-50 parts of gamma-aminobutyric acid, 4-30 parts of oxalic acid sodium salt and 200-530 parts of gamma-glutamyl-phenylalanine;

The preparation method comprises the steps of washing cotton stalks and cornstalks with distilled water, sun-drying, crushing the cotton stalks and the cornstalks to a mixture of 80-120 meshes, adding Mandel salt solution into the mixture, inoculating the mixture into mixed bacterial liquid of aspergillus oryzae and bacillus amyloliquefaciens, adjusting the pH value of the mixed liquid to 8-10, culturing for 2-4 days at a constant temperature of 35-50 ℃, centrifuging the obtained enzyme fermentation liquid for 15-30 min at a speed of 1000-1300 r/min, and taking supernatant to be subjected to rotary evaporation at a temperature of 105-150 ℃.

The composition comprises, by weight, 10-45 parts of berberine, 15-40 parts of allicin, 8-25 parts of benfotiamine, 10-43 parts of gamma-aminobutyric acid, 7-25 parts of oxalic acid sodium salt and 260-500 parts of gamma-glutamyl-phenylalanine.

The composition further preferably comprises, by weight, 20-40 parts of berberine, 17-35 parts of allicin, 10-20 parts of benfotiamine, 15-40 parts of gamma-aminobutyric acid, 10-20 parts of oxalic acid sodium salt and 300-470 parts of gamma-glutamyl-phenylalanine.

More preferably, the composition comprises, by weight, 23-38 parts of berberine, 20-33 parts of allicin, 13-18 parts of benfotiamine, 30-39 parts of gamma-aminobutyric acid, 12-16 parts of oxalic acid sodium salt and 350-450 parts of gamma-glutamyl-phenylalanine.

The preparation method of the fishing regulation and control composition with the functions of reducing blood lipid and controlling blood sugar comprises the following specific steps:

S1, respectively washing cotton stalks and cornstalks with distilled water, sun-drying, crushing the cotton stalks and cornstalks to a mixture with 80-120 meshes, adding Mandel salt solution into the mixture, then inoculating the mixture into mixed bacterial liquid of aspergillus oryzae and bacillus amyloliquefaciens, regulating the pH value of the mixed liquid to 8-10, culturing for 2-4 days at a constant temperature of 35-50 ℃, centrifuging the obtained enzyme fermentation liquid for 15-30 min at a speed of 1000-1300 r/min, and taking supernatant liquid for rotary evaporation at a temperature of 105-150 ℃ to obtain gamma-glutamyl-phenylalanine;

S2, respectively crushing berberine, garlicin, benfotiamine, gamma-aminobutyric acid, oxalic acid sodium salt and gamma-glutamyl-phenylalanine, and then sieving with a 120-150 mesh sieve and fully and uniformly mixing;

S3, adding beta-cyclodextrin and a dispersing agent into a methanol aqueous solution, homogenizing for 10-15 min at 10000-12000 r/min, adding a 10-15% methanol solution of the regulating composition under the stirring condition of 1000-1300 r/min, stirring for 4-5 h at 30-65 ℃ and 1000-1300 r/min, naturally cooling to room temperature, standing for 24-36 h, carrying out suction filtration, washing, cooling and drying to obtain the fishing regulating composition with the functions of reducing blood lipid and controlling blood sugar.

Preferably, in the step S1, the mass ratio of the cotton stalks to the corn stalks is (1-2.5): (3-5.4), the mass ratio of the aspergillus oryzae and the bacillus amyloliquefaciens in the mixed bacterial liquid is (2.5-3): (1-2.5), and the volume ratio of the mass ratio of the mixture to the Mandel salt solution is (1-2) g (2-5) mL.

Preferably, the temperature of berberine, allicin, benfotiamine, gamma-aminobutyric acid, oxalic acid sodium salt and gamma-glutamyl-phenylalanine in step S2 is less than 40 ℃.

Preferably, in the step S3, the dispersing agent is a mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1, the methanol aqueous solution is a mixed solution of methanol and distilled water in a volume ratio of 1:3, the mass ratio of the beta-cyclodextrin to the dispersing agent is 3 (3.5-5), the volume ratio of the total mass of the beta-cyclodextrin and the dispersing agent to the methanol aqueous solution is (400-500) g (1-2.5) L, and the volume ratio of the methanol aqueous solution to the methanol solution of the regulating composition is 20 (1-1.5).

The fishing regulation and control composition with the functions of reducing blood lipid and controlling blood sugar is applied to freshwater fish culture.

Preferably, the addition amount of the regulation and control composition in the freshwater fish feed is 0.5-1.4wt%, and the freshwater fish is grass carp, megalobrama amblycephala, carp, crucian, tilapia mossambica, channel catfish, larch or snakehead.

According to the invention, cotton stalks and corn stalks are mixed in proportion and then added into a composite strain for fermentation, so that the gamma-glutamyl-phenylalanine with high purity is obtained, and then the gamma-glutamyl-phenylalanine is used for preparing the fishing regulation and control composition. The process can greatly improve the comprehensive utilization of crop straw resources, and has great significance for promoting the income increase of farmers, protecting the environment, saving resources and sustainable development of agricultural economy. Meanwhile, in order to further improve the bioavailability of the composition and the effect thereof, a microencapsulation technique is applied to the preparation process of the composition. Through technical optimization and culture experiment screening, the fishing regulation and control composition with strong phagostimulant is compatible based on pharmacological synergistic relationship of benfotiamine, berberine, garlicin, gamma-aminobutyric acid and sodium oxalate. The feed additive is used as an efficient environment-friendly aquatic feed additive, can greatly reduce the use amount of protein and fat raw materials in the feed, and remarkably improves the economic benefit of aquaculture.

Compared with the prior art, the invention has the following beneficial effects:

1. The regulation and control composition takes berberine, allicin, benfotiamine, gamma-aminobutyric acid and oxalic acid sodium salt as main raw materials, and fully utilizes the characteristics of different biological structures to carry out scientific compatibility so as to play a synergistic effect. Meanwhile, plant straw resources are fully utilized, and the strong food gamma-glutamyl-phenylalanine is produced by means of a biological fermentation technology and added into the regulation and control composition. In addition, microencapsulation technology is also applied to the processing of the conditioning composition to enhance the bioavailability of the aquatic animal.

2. The regulation composition achieves the effects of reducing blood lipid and controlling blood sugar by regulating and controlling the glycolipid metabolism function of aquatic animals, thereby improving the carbohydrate metabolism function of the aquatic animals and increasing the utilization rate of carbohydrates in feed.

Drawings

FIG. 1 is the effect of different feeds on the weight gain rate of young megalobrama amblycephala in application example 1;

FIG. 2 is the effect of different feeds on the weight gain rate of young megalobrama amblycephala in application example 2;

FIG. 3 shows the effect of different feeds on liver fat content of young megalobrama amblycephala in application example 2;

FIG. 4 is the effect of different feeds on blood glucose levels of young megalobrama amblycephala in application example 2;

FIG. 5 is the effect of different feeds on blood glucose levels of carp in use example 4;

FIG. 6 is the effect of different feeds on liver fat content of young micropterus salmoides in application example 7;

FIG. 7 is the effect of different feeds on the blood glucose clearance of snakeheads in application example 9;

FIG. 8 is a graph showing the dynamic change of blood glucose of snakehead with different feeds in application example 9.

Detailed Description

The present invention is further illustrated below in conjunction with specific examples, but should not be construed as limiting the invention. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.

The Mandel salt solution in the embodiment of the invention comprises a macroelement solution and a microelement solution, wherein the macroelement solution is prepared by adding K₂HPO₄2.0g,(NH₄)₂SO₄1.4g,MgSO₄·7H₂O 0.3g,CaCl₂0.3g mL of double distilled water to dissolve the macroelement solution, the microelement solution is prepared by adding FeSO₄·7H₂O 5.0mg,MnSO₄·H₂O1.6mg,ZnSO₄·7H₂O 1.4mg,CoCl₂2.0mg, mL of double distilled water to dissolve the double distilled water, mixing the macroelement solution and the microelement solution, adjusting the pH value to 5.5-6.0 by using HCl, and fixing the volume to 1000mL by using the double distilled water to obtain Mandel salt solution.

Example 1

1. Cotton stalks and cornstalks are respectively washed and dried in the sun by distilled water, crushed to 100 meshes, and 10g of mixture (the mass ratio of the cotton stalks to the cornstalks is 1:3) is taken and added with 20mL Mandel salt solution. Then, a mixed bacterial solution of aspergillus oryzae and bacillus amyloliquefaciens (the mass ratio of the aspergillus oryzae to the bacillus amyloliquefaciens is 2.5:1) is inoculated, the mixed bacterial solution is cultured for 2 days in a constant temperature incubator with the pH adjusted to 8.5,45 ℃, the obtained enzyme fermentation liquid is centrifuged for 20min at 1300r/min, and the supernatant is taken for 105 ℃ rotary evaporation, so that gamma-glutamyl-phenylalanine is obtained.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 2g of berberine, 4g of allicin, 2g of benfotiamine, 2g of sodium oxalate salt, 4g of gamma-aminobutyric acid and 11g of gamma-glutamyl-phenylalanine which are respectively screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process needs to ensure that the mixture is uniform and stable, so that 25g of coarse control composition is obtained.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 10000r/min for 10min. 100mL of 10wt% strength by mass methanol solution of the control composition was added under 1300r/min stirring, stirred at 50℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of a fishing control composition.

Example 2

1. Cotton stalks and corn stalks are washed and dried in the sun respectively by distilled water, crushed to 120 meshes, and 13g of mixture (the mass ratio of the cotton stalks to the corn stalks is 2.5:3) is taken and added with 50mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (the mass ratio of the Aspergillus oryzae to the Bacillus amyloliquefaciens is 3:2) is inoculated, the pH is regulated to 10.0, the mixture is cultured in a 50 ℃ constant temperature incubator for 4 days, the obtained enzyme fermentation broth is centrifuged at 1000r/min for 30min, and the supernatant is taken for rotary evaporation at 150 ℃ to obtain gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 3g of berberine, 3g of allicin, 4g of benfotiamine, 3g of sodium oxalate salt, 2g of gamma-aminobutyric acid and 10g of gamma-glutamyl-phenylalanine which are screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process needs to ensure that the mixture is uniform and stable, so that 25g of coarse control composition is obtained.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 12000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under 1200r/min stirring, stirred at 65℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 3

1. Cotton stalks and cornstalks are respectively washed and dried in the sun by distilled water, crushed to 80 meshes, and 20g of mixture (the mass ratio of the cotton stalks to the cornstalks is 2:5) is taken and added into 43mL Mandel salt solution. Subsequently, a mixed bacterial solution of aspergillus oryzae and bacillus amyloliquefaciens (mass ratio of aspergillus oryzae and bacillus amyloliquefaciens=3:1.5) was inoculated, the pH was adjusted to 9.0, and the culture was carried out at a constant temperature of 40 ℃ for 4 days. Finally, the enzyme fermentation broth is centrifuged for 25min at 1200r/min, and the supernatant is taken to obtain the gamma-glutamyl-phenylalanine after rotary evaporation at 145 ℃.

2. When the berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are crushed, the temperature is less than 40 ℃, 1g of berberine, 4g of allicin, 3g of benfotiamine, 6g of sodium oxalate salt, 1g of gamma-aminobutyric acid and 10g of gamma-glutamyl-phenylalanine which are screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process needs to ensure that the mixture is uniform and stable, so that 25g of coarse adjustment control composition is obtained.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 12000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 60℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 4

1. Cotton stalks and corn stalks are washed and dried in the sun respectively by distilled water, crushed to 120 meshes, and 17g of mixture (the mass ratio of the cotton stalks to the corn stalks is 1.5:3.5) is taken and added into 40mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.3:2) was inoculated, pH was adjusted to 8.0, and the mixture was cultured in a 45℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1200r/min for 30min, and the supernatant was subjected to rotary evaporation at 150℃to obtain gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 5g of berberine, 1g of allicin, 1g of benfotiamine, 2g of sodium oxalate salt, 7g of gamma-aminobutyric acid and 9g of gamma-glutamyl-phenylalanine which are screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process is required to ensure that the mixture is uniform and stable, so as to obtain 25g of coarse adjustment control composition.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 11000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 60℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 5

1. Cotton stalks and corn stalks are washed and dried in the sun respectively by distilled water, crushed to 120 meshes, and 15g of mixture (the mass ratio of the cotton stalks to the corn stalks is 2.5:4.0) is taken and added into 40mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.5:1) was inoculated, pH was adjusted to 9.5, and the mixture was cultured in a 50℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1000r/min for 30 minutes, and the supernatant was subjected to rotary evaporation at 140℃to obtain gamma-glutamyl-phenylalanine.

2. When the berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are crushed, the temperature is less than 40 ℃, 1g of berberine, 1g of allicin, 4g of benfotiamine, 4g of sodium oxalate salt, 1g of gamma-aminobutyric acid and 14g of gamma-glutamyl-phenylalanine which are screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process needs to ensure that the mixture is uniform and stable, so that 25g of coarse adjustment control composition is obtained.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 11000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 55℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 6

1. Cotton stalks and corn stalks are washed and dried in the sun respectively by distilled water, crushed to 120 meshes, and 15g of mixture (the mass ratio of the cotton stalks to the corn stalks is 2.5:4.5) is taken and added into 50mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.5:2) was inoculated, pH was adjusted to 10.0, and the mixture was cultured in a 40℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1300r/min for 30min, and the supernatant was subjected to rotary evaporation at 140℃to obtain gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 4g of berberine, 8g of allicin, 1g of benfotiamine, 1g of sodium oxalate salt, 3g of gamma-aminobutyric acid and 8g of gamma-glutamyl-phenylalanine which are screened by a 120-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process is required to ensure that the mixture is uniform and stable, so as to obtain 25g of coarse adjustment control composition.

Example 7

1. Cotton stalks and corn stalks are washed and dried in the sun respectively by distilled water, crushed to 120 meshes, and 20g of mixture (the mass ratio of the cotton stalks to the corn stalks is 2.5:5) is taken and added into 30mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2:1) was inoculated, pH was adjusted to 10.0, and the mixture was cultured in a 35℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1300r/min for 30 minutes, and the supernatant was subjected to rotary evaporation at 150℃to obtain gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 2g of berberine, 1g of allicin, 2g of benfotiamine, 1g of sodium oxalate salt, 3g of gamma-aminobutyric acid and 16g of gamma-glutamyl-phenylalanine which are screened by a 150-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process is required to ensure that the mixture is uniform and stable, so as to obtain 25g of coarse adjustment control composition.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 11000r/min for 10min. 100mL of 10wt% strength by mass methanol solution of the control composition was added with stirring at 1300r/min, stirred at 62℃for 4.5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 8

1. Cotton stalks and cornstalks are respectively washed and dried in the sun by distilled water, crushed to 120 meshes, and 20g of mixture (the mass ratio of the cotton stalks to the cornstalks is 1.5:3) is taken and added into 50mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.6:1.5) was inoculated, and the mixture was cultured in an incubator at a pH of 10.0,50 ℃for 4 days. Finally, the enzyme fermentation broth is centrifuged for 25min at 1000r/min, and the supernatant is taken for rotary evaporation at 150 ℃ to obtain the gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 3g of berberine, 3g of allicin, 4g of benfotiamine, 4g of sodium oxalate salt, 1g of gamma-aminobutyric acid and 10g of gamma-glutamyl-phenylalanine which are screened by a 150-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process is required to ensure that the mixture is uniform and stable, so as to obtain 25g of coarse adjustment control composition.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 12000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 62℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 9

1. Cleaning cotton stalks and corn stalks with distilled water, sun-drying, crushing to 120 meshes, and adding 10g of mixture (the mass ratio of the cotton stalks to the corn stalks is 2.5-5.0) into 50mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.5:1.0) was inoculated, pH was adjusted to 10.0, and the mixture was cultured in a 50℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1200r/min for 30min, and the supernatant was subjected to rotary evaporation at 150℃to obtain gamma-glutamyl-phenylalanine.

2. The berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are all required to be smaller than 40 ℃ when being crushed, 3g of berberine, 3g of allicin, 2g of benfotiamine, 2g of sodium oxalate salt, 8g of gamma-aminobutyric acid and 7g of gamma-glutamyl-phenylalanine which are screened by a 130-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process is required to ensure that the mixture is uniform and stable, so as to obtain 25g of coarse adjustment control composition.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2.5L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 12000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 60℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Example 10

1. Cotton stalks and cornstalks are respectively washed and dried in the sun by distilled water, crushed to 120 meshes, and 20g of mixture (the mass ratio of the cotton stalks to the cornstalks is 2.5:3) is taken and added with 0mLMandel salt solution. Subsequently, a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens (mass ratio of Aspergillus oryzae and Bacillus amyloliquefaciens=2.8:1.5) was inoculated, pH was adjusted to 10.0, and the mixture was cultured in a 50℃incubator for 4 days, and the obtained enzyme fermentation broth was centrifuged at 1300r/min for 30min, and the supernatant was subjected to rotary evaporation at 150℃to obtain gamma-glutamyl-phenylalanine.

2. When the berberine, the allicin, the benfotiamine, the sodium oxalate salt, the gamma-aminobutyric acid and the gamma-glutamyl-phenylalanine are crushed, the temperature is less than 40 ℃, 1g of berberine, 3g of allicin, 2g of benfotiamine, 2g of sodium oxalate salt, 4g of gamma-aminobutyric acid and 13g of gamma-glutamyl-phenylalanine which are screened by a 150-mesh sieve are fully and uniformly mixed according to a step-by-step uniform mixing method, and the process needs to ensure that the mixture is uniform and stable, so that 25g of coarse adjustment control composition is obtained.

3. 300G of beta-cyclodextrin and 500g of dispersant (mixture of calcium lignosulfonate and polyvinylpyrrolidone K17 in a weight ratio of 2:1) were added to 2L of aqueous methanol solution (mixed solution of methanol and distilled water in a volume ratio of 1:3) and homogenized at 12000r/min for 10min. 100mL of 10wt% methanol solution of the control composition was added under stirring at 1300r/min, stirred at 65℃for 5h, cooled naturally to room temperature and allowed to stand for 24h. Then, the mixture was suction-filtered and washed with a Buchner funnel, and cooled and dried to obtain 10g of the control composition.

Application example 1

The young megalobrama amblycephala (about 140 g) cultivation test and result analysis are carried out on an aquatic product teaching and research base of Nanjing agricultural university (Nanjing Shikou district Starfish town). The young megalobrama amblycephala is purchased from national aquatic fine-breed farms in Yangzhou city of Jiangsu province. 360 young fishes (initial weight: 143.12 +/-2.10 g) with regular specifications are selected and randomly placed into 12 net cages (specification: 2.0m×1.0 m), and each net cage is provided with 30 fish. Wherein, (1) the control group was fed with basal feed (feed sugar level was 35%), (2) the high sugar group was fed with high sugar feed (feed sugar level was 45%), (3) the high sugar + regulatory composition group was fed with high sugar feed +1.0% of the regulatory composition prepared in example 1. The feed formulation components are shown in table 1. Feeding and growing conditions of young megalobrama amblycephala are observed during 12 weeks of cultivation, blood and tissue samples are collected after the cultivation is finished, and subsequent index analysis is carried out, and specific results are shown in table 2.

Table 1 test feed formulation

TABLE 2 influence of different feeds on growth and blood Biochemical index of young megalobrama amblycephala

Note that the same lowercase label on the same row indicates that the difference is not significant.

FIG. 1 shows the effect of different feeds on the weight gain rate of young megalobrama amblycephala in application example 1. The same lowercase label in the figure indicates that the difference is not significant. As shown in fig. 1 and table 2, the weight gain rate, islet beta cell function and insulin sensitivity index of the fish in the high-sugar group feed were significantly lower than those in the control group (P < 0.05), but the liver body ratio, abdominal fat rate, liver glycogen and fat content, blood sugar level, insulin content, glutamate oxaloacetate activity, glutamate pyruvate transaminase activity, plasma pro-inflammatory cytokines (tumor necrosis factor, interleukin 1 beta and interleukin 6) levels, liver pro-inflammatory cytokines (interleukin 1 beta and interleukin 6) content and insulin resistance index were significantly higher than those in the control group (P < 0.05). The feed of the high sugar + regulatory composition increases the rate of weight gain, islet beta cell function and insulin sensitivity in fish, but the liver mass ratio, abdominal fat rate, liver glycogen content, blood glucose level, glutamate oxaloacetate transaminase activity, glutamate pyruvate transaminase activity, plasma pro-inflammatory cytokine (tumor necrosis factor, interleukin 1 beta and interleukin 6) levels, liver pro-inflammatory cytokine (interleukin 1 beta and interleukin 6) content and insulin resistance index are significantly lower than (P < 0.05) the high sugar group.

The results show that the feed containing the regulation and control composition prepared in the embodiment 1 can improve the phenomena of excessive accumulation of liver fat, liver inflammatory reaction and the like caused by feeding high-sugar feed, strengthen the pancreatic islet beta cell function of the fish body, promote the secretion and release of insulin, increase the insulin sensitivity and reduce the insulin resistance degree, and finally reduce the blood sugar level and improve the utilization rate of saccharides in the feed by the megalobrama amblycephala. In addition, the use of sugar metabolism regulators significantly reduces hepatic glycogen deposition and prevents the occurrence of liver glycolipid metabolic disorders due to hepatomegaly.

Application example 2

The breeding test and result analysis of young megalobrama amblycephala (about 50 g) are carried out in an indoor circulating breeding system of zootechnical building of agricultural university in south China. The young megalobrama amblycephala fish is purchased from aquatic breeding company in Guangzhou city. Selecting 960 young fishes (initial weight: 53.31 + -3.03 g) with regular specification, randomly placing into 32 net cages (specification: 2.0mX1.0mX1.0m), and each net cage comprises 30 tail fishes. Wherein, (1) HC, high sugar group, high sugar feed (feed sugar level 45%), (2) HC1, high sugar +1.0% berberine group, (3) HC2, high sugar +1.0% allicin group, (4) HC3, high sugar +1.0% benfotiamine group, (5) HC4, high sugar +1.0% oxalic acid sodium salt group, (6) HC5, high sugar +1.0% gamma-aminobutyric acid group, (7) HC6, high sugar +1.0% gamma-glutamyl-phenylalanine, and (8) HC7, high sugar +regulatory composition group, high sugar feed +1.0% regulatory composition prepared in example 1. The basic components of the feed formula are shown in table 1. And (3) culturing for 12 weeks, observing feeding and growth conditions of young megalobrama amblycephala, and collecting blood and tissue samples after culturing is finished for subsequent index analysis, wherein the specific results are shown in figures 2-4.

FIG. 2 is the effect of different feeds on the weight gain rate of young megalobrama amblycephala in application example 2, FIG. 3 is the effect of different feeds on the liver fat content of young megalobrama amblycephala in application example 2, and FIG. 4 is the effect of different feeds on the blood sugar level of young megalobrama amblycephala in application example 2. The same lowercase label in the figure indicates that the difference is not significant. As can be seen from fig. 2-4, the weight gain rate of the fish in the high-sugar group feed was significantly lower than that of the other test group fish (P < 0.05), but the trend of the change of the liver fat content and the blood sugar level was opposite to that (P < 0.05). In the high-sugar feed additive group, the feed of the high-sugar + regulation composition significantly increases the weight gain rate of fish, but the liver fat content and the blood sugar level are significantly lower (P < 0.05) than those of each high-sugar feed additive group fish. The results show that compared with each single regulator, the regulating composition is more beneficial to promoting the growth of fish which ingests high-sugar feed, and has obvious lipid-lowering and sugar-controlling effects.

Application example 3

The grass carp cultivation test and result analysis are carried out in an indoor circulating cultivation system of special economic animals and aquatic systems of Nanjing agricultural university. Grass carp with average weight of about 42g is selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar plus regulatory composition group. Each group of 4 circulating culture tanks (specification: 480L), each tank contains 30 fish, and the culture period is 12 weeks. The feed formulation is shown in Table 1. After the cultivation test is finished, blood and liver samples are collected for index measurement.

As can be seen from table 3, after 12 weeks of culture test, the weight gain rate, blood sugar clearance ability and insulin sensitivity index of the high-sugar group fish were significantly lower than those of the control group (P < 0.05), while the liver mass ratio, liver glycogen content, liver fat content, blood sugar level, plasma pro-inflammatory cytokine (tumor necrosis factor, interleukin 1 beta and interleukin 6) level, blood sugar half-life and liver pro-inflammatory cytokine (tumor necrosis factor and interleukin 1 beta) expression levels were all significantly increased (P < 0.05), indicating that long-term feeding of the high-sugar group fish can cause liver fat accumulation of fish body and produce liver inflammatory reaction, reduce tissue insulin sensitivity, and further lower blood sugar regulation ability, and the weight gain rate was lowered. However, the high sugar + regulatory composition group fish has significantly lower liver glycogen content, liver fat content, blood glucose level, plasma pro-inflammatory cytokine (tumor necrosis factor, interleukin 1 beta and interleukin 6) level, blood glucose half-life and expression level of liver pro-inflammatory cytokine than the high sugar group (P < 0.05), but has increased weight gain rate and plasma insulin level. This shows that the feed containing the regulation composition prepared in example 1 can improve liver lipid accumulation and inflammatory reaction of grass carp caused by ingestion of high-sugar feed, further enhance blood sugar removing capacity of the fish body, reduce blood sugar half-life, promote insulin secretion, and finally increase weight gain rate and economic benefit of cultivation.

TABLE 3 influence of different feeds on grass carp growth Properties and blood Biochemical index

Application example 4

The carp breeding test and result analysis are carried out in an indoor circulating breeding system of zootechnical building of agricultural university in south China. Carps weighing about 51g in average were selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar + regulatory composition group. Each group of 4 circulating culture tanks (specification: 500L), each tank is used for 30 fish, and the culture period is 12 weeks. The feed formulation is as shown in table 1, but 1.0% of the conditioning composition of example 2 is used. After the cultivation test is finished, blood and liver samples are collected for index measurement.

FIG. 5 shows the effect of different feeds on blood glucose levels of carp in application example 4, where the same lowercase notations indicate insignificant differences. As shown in table 4 and fig. 5, after 12 weeks of culture test, the weight gain rate, the blood sugar clearance capacity and the insulin sensitivity index of the high-sugar group fish are significantly lower than those of the control group (P < 0.05), and the abdominal fat rate, the liver fat content, the blood sugar level and the blood sugar half-life of the high-sugar group fish are all significantly increased (P < 0.05), which indicates that long-term feeding of the high-sugar feed can lead to liver fat accumulation of fish body, reduce tissue insulin sensitivity, further reduce blood sugar regulation capacity and significantly reduce the weight gain rate. However, the liver fat content, blood glucose level, blood glucose half-life of the fish in the group of high sugar + regulatory compositions were significantly lower than in the high sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the regulatory composition prepared in example 2 can improve liver lipid accumulation of carp caused by ingestion of high sugar feed, enhance blood sugar removing ability of fish body and promote insulin secretion, thereby being beneficial to increasing weight gain rate of fish body.

TABLE 4 influence of different feeds on growth Properties and Biochemical index of carp

Application example 5

The crucian cultivation test and the result analysis are carried out in an indoor circulating cultivation system of the zootechnical building of the agricultural university in south China. The crucian carp with average weight of about 10g is selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar plus regulation composition group. Each group of 4 circulating culture tanks (specification: 500L), each tank is used for 30 fish, and the culture period is 12 weeks. The feed formulation is as shown in table 1, but 1.0% of the conditioning composition of example 5 is used. After the cultivation test is finished, blood and liver samples are collected for index measurement.

As shown in table 5, after 12 weeks of culture test, the weight gain rate, blood sugar clearance ability and insulin sensitivity index of the high-sugar group fish were significantly lower than those of the control group (P < 0.05), while the abdominal fat rate, liver fat content, blood sugar level and blood sugar half-life of the high-sugar group fish were all significantly increased (P < 0.05), indicating that the fish fed with the high-sugar feed had liver fat accumulation and tissue insulin sensitivity was reduced, thereby reducing blood sugar regulation ability and weight gain rate was significantly reduced. However, the liver fat content, blood glucose level, blood glucose half-life of the fish in the group of high sugar + regulatory compositions were significantly lower than in the high sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the regulation composition prepared in example 5 can improve liver lipid accumulation of crucian, enhance blood sugar removing ability of the fish body and promote insulin secretion, thereby being beneficial to improving weight gain rate of the fish body and economic benefit of cultivation.

TABLE 5 influence of different feeds on growth performance and Biochemical index of Carassius auratus

Application example 6

The tilapia culture test and result analysis are carried out in an indoor circulating culture system of the dynamic building of the agricultural university in south China. Tilapia weighing about 80g was selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar + regulatory composition group. Each group of 4 circulating culture tanks (specification: 500L), each tank is used for 30 fish, and the culture period is 12 weeks. The feed formulation is as shown in table 1, but 1.2% of the conditioning composition of example 4 is used. After the cultivation test is finished, blood and liver samples are collected for index measurement.

As shown in table 6, after 12 weeks of culture test, the weight gain rate of the high-sugar group fish was significantly lower than that of the control group (P < 0.05), while the abdominal fat rate, liver fat content, and blood sugar level of the high-sugar group fish were all significantly increased (P < 0.05), indicating that the fish fed the high-sugar feed had liver fat accumulation, postprandial hyperglycemia symptoms, and the weight gain rate was significantly reduced. However, the liver fat content and blood glucose levels of the fish in the high-sugar + regulatory composition group were significantly lower than those in the high-sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the control composition prepared in example 4 can improve liver lipid accumulation of tilapia caused by feeding high-sugar feed, enhance the blood sugar regulating capability of the tilapia body, and further improve the weight gain rate of the cultured tilapia.

TABLE 6 influence of different feeds on growth performance and Biochemical index of Tilapia

Application example 7

The largemouth black bass cultivation test and result analysis are carried out in an indoor circulating cultivation system of the zootechnical building of the agricultural university of south China. The largehead jewfish with a mean weight of about 5g was selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar + regulatory composition group. Each group of 4 circulating culture tanks (specification: 500L), 20 fish in each tank, and the culture period is 12 weeks. The feed formulation is shown in table 7. After the cultivation test is finished, blood and liver samples are collected for index measurement.

FIG. 6 is the effect of different feeds on liver fat content of young micropterus salmoides in application example 7. The same lowercase label in the figure indicates that the difference is not significant. As shown in table 8 and fig. 6, the weight gain rate, blood sugar clearance and insulin sensitivity index of the high-sugar group fish were significantly lower than those of the control group (P < 0.05), while the abdominal fat rate, liver fat content, blood sugar level and blood sugar half-life of the high-sugar group fish were all significantly increased (P < 0.05), indicating that the liver fat accumulation, tissue insulin sensitivity decrease and blood sugar continuous high phenomena occurred in the larch fed with the high-sugar feed, but the weight gain rate was significantly decreased. However, the liver fat content, blood glucose level, blood glucose half-life of the fish in the group of high sugar + regulatory compositions were significantly lower than in the high sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the regulatory composition prepared in example 5 can improve liver lipid accumulation of micropterus salmoides caused by ingestion of high-sugar feed, enhance blood sugar removing ability of fish bodies and promote insulin secretion, thereby increasing weight gain rate of fish bodies.

Table 7 test feed formulation

TABLE 8 influence of different feeds on growth performance and Biochemical index of Lateolabrax

Application example 8

And (3) carrying out a channel catfish culture test and result analysis, wherein the culture test is carried out in an indoor circulating culture system of an animal building of the agricultural university of south China. Channel catfish with a mean weight of about 15g was selected and randomly split into three experimental groups, namely a control group, a high sugar group and a high sugar + regulatory composition group. Each group of 4 circulating culture tanks (specification: 500L), 25 fish in each tank, and the culture period is 12 weeks. The feed formulation is shown in table 7, but 2.0% of the conditioning composition of example 6 is used. After the cultivation test is finished, blood and liver samples are collected for index measurement.

As shown in table 9, after 12 weeks of culture test, the weight gain rate of the high-sugar group fish was significantly lower than that of the control group (P < 0.05), while the abdominal fat rate, liver fat content and blood glucose level of the high-sugar group channel catfish were all significantly increased (P < 0.05), indicating that the channel catfish fed with the high-sugar feed had liver fat accumulation, postprandial hyperglycemia symptoms and significantly decreased weight gain rate. However, the liver fat content and blood glucose levels of the fish in the high-sugar + regulatory composition group were significantly lower than those in the high-sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the regulatory composition prepared in example 6 can improve liver lipid accumulation of channel catfish caused by ingestion of high-sugar feed, enhance blood sugar regulation capability of fish body and increase weight gain rate of cultured tilapia.

TABLE 9 Effect of different feeds on growth Performance and Biochemical index of Tilapia

Application example 9

And (3) performing snakehead culture tests and result analysis, wherein the culture tests are performed in an indoor circulating culture system of the dynamic building of the agricultural university of south China. Snakehead with average weight of about 35g is selected and randomly divided into three experimental groups, namely a control group, a high sugar group and a high sugar plus regulatory composition group. Each group of 4 circulating culture tanks (specification: 500L), each tank is used for 30 fish, and the culture period is 12 weeks. The feed formulation is shown in table 7, but 1.3% of the conditioning composition of example 8 is used. After the cultivation test is finished, blood and liver samples are collected for index measurement.

The remaining fish were subjected to glucose tolerance experiments by starving the fish for 24 hours and then anesthetizing the fish, then, intraperitoneally injecting a glucose solution of 1.67g/kg body weight, after the injection was completed, putting the fish into 28 indoor circulation water tanks, namely, 5 experimental groups, 7 tanks (corresponding to 7 sampling points) each, and then, collecting blood samples at 0, 1,2, 4, 8 and 12 hours, respectively. A cylinder was taken at each time point of each group to avoid the influence of density variations on it and to reduce the stress on the fish body caused by repeated sampling.

FIG. 7 shows the effect of different feeds on the glycemic clearance of snakeheads according to application example 9. The same lowercase label in the figure indicates that the difference is not significant. As shown in table 10 and fig. 7, the weight gain rate, blood sugar clearance and insulin sensitivity index of the fish fed with the high sugar group feed were significantly lower than those of the control group (P < 0.05), while the abdominal fat rate, liver fat content, blood sugar level and blood sugar half-life of the fish fed with the high sugar group feed were all significantly increased (P < 0.05), indicating that the snakehead fed with the high sugar group feed had the phenomena of liver fat accumulation, tissue insulin sensitivity reduction and blood sugar continuous elevation, but the weight gain rate was significantly reduced. However, the liver fat content, blood glucose level, blood glucose half-life of the fish in the group of high sugar + regulatory compositions were significantly lower than in the high sugar group (P < 0.05), but the rate of gain and plasma insulin levels were elevated. This shows that the feed containing the control composition prepared in example 8 can improve liver lipid accumulation of snakeheads caused by feeding high-sugar feed, enhance blood sugar removing capacity of fish bodies and promote insulin secretion, thereby improving weight gain rate of fish bodies.

TABLE 10 influence of different feeds on growth performance and Biochemical index of snakeheads

FIG. 8 is a graph showing the dynamic change of blood glucose of snakeheads by different feeds in application example 9, and the effect of the different feeds on glucose tolerance of snakeheads was studied. As can be seen from fig. 8, the maximum value of the plasma glucose level of each test group fish appeared at 2h after glucose loading, and showed a decreasing trend with increasing sampling time. The blood glucose level of the fish in the high sugar + regulatory composition group was significantly lower than that of the other test groups throughout the sampling time and returned to the basal value at 12 h. This indicates that the high sugar + regulatory composition group feed can significantly enhance the glucose tolerance of snakeheads.

In conclusion, the invention combines the cultivation test to deeply explore the proper effect and dosage of the regulation and control composition in the feed and the regulation and control mechanism of the regulation and control composition on the sugar metabolism functions of herbivory (grass carp and megalobrama amblycephala), omnivorous (carp, crucian, tilapia and channel catfish) and carnivorous (larch and snakehead) fishes in different growth stages. Therefore, the method has strong pertinence to the regulation of the sugar metabolism function of aquatic animals.

The invention combines the results of growth and physical and chemical indexes of related culture experiments, and proves that the regulation and control composition can obviously improve lipid accumulation and inflammatory response of fish metabolic tissues induced by high-sugar feed, strengthen the glycolipid metabolism function of the fish, obviously enhance the insulin synthesis and secretion capacity of the fish, reduce postprandial blood sugar level of the fish, relieve metabolic pressure of the fish body, obviously improve the sugar tolerance capacity of the fish, improve the utilization rate of sugar in the feed, further reduce the feed coefficient and increase the culture economic benefit.

The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims

1. A fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions, characterized in that the composition comprises the following components in parts by weight: 2-50 parts of berberine; 4-50 parts of allicin; 2-30 parts of benfotiamine; 2-50 parts of γ-aminobutyric acid; 4-30 parts of sodium oxalate; 200-530 parts of γ-glutamyl-phenylalanine;

Among them, γ-glutamyl-phenylalanine is prepared by washing cotton stalks and corn stalks with distilled water, drying them, and then crushing them into a mixture of 80-120 mesh. The mixture is added with Mandel salt solution and then inoculated into a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens. The pH of the mixed solution is adjusted to 8-10, and the mixture is cultured at a constant temperature of 35-50°C for 2-4 days. The obtained enzyme fermentation solution is centrifuged at 1000-1300r/min for 15-30min, and the supernatant is rotary evaporated at 105-150°C to obtain the product.

2. The fishery regulating composition with lipid-lowering and blood sugar-controlling functions according to claim 1, characterized in that the composition comprises the following components in parts by weight: 10 to 45 parts of berberine; 15 to 40 parts of allicin; 8 to 25 parts of benfotiamine; 10 to 43 parts of γ-aminobutyric acid; 7 to 25 parts of sodium oxalate; and 260 to 500 parts of γ-glutamyl-phenylalanine.

3. The fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to claim 2 is characterized in that the composition comprises the following components in parts by weight: 20 to 40 parts of berberine; 17 to 35 parts of allicin; 10 to 20 parts of benfotiamine; 15 to 40 parts of γ-aminobutyric acid; 10 to 20 parts of sodium oxalate; and 300 to 470 parts of γ-glutamyl-phenylalanine.

4. The fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to claim 3 is characterized in that the composition comprises the following components in parts by weight: 23-38 parts of berberine; 20-33 parts of allicin; 13-18 parts of benfotiamine; 30-39 parts of γ-aminobutyric acid; 12-16 parts of sodium oxalate; and 350-450 parts of γ-glutamyl-phenylalanine.

5. The method for preparing the fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to any one of claims 1 to 4, characterized in that it comprises the following specific steps:

S1. Wash cotton stalks and corn stalks with distilled water, dry them in the sun, and then grind them into a mixture of 80-120 mesh. Add Mandel salt solution to the mixture and then inoculate it into a mixed bacterial solution of Aspergillus oryzae and Bacillus amyloliquefaciens. Adjust the pH of the mixture to 8-10, culture it at a constant temperature of 35-50°C for 2-4 days, centrifuge the obtained enzyme fermentation liquid at 1000-1300r/min for 15-30min, take the supernatant and rotary evaporate it at 105-150°C to obtain γ-glutamyl-phenylalanine;

S2. After berberine, allicin, benfotiamine, γ-aminobutyric acid, sodium oxalate and γ-glutamyl-phenylalanine were crushed separately, passed through a 120-150 mesh sieve and then fully mixed to obtain a coarsely controlled composition;

S3. Add β-cyclodextrin and dispersant to methanol aqueous solution and homogenize at a speed of 10000-12000 r/min for 10-15 min. Add methanol solution of the crude regulating composition with a mass concentration of 10-15 wt% under stirring at 1000-1300 r/min. Stir at 30-65°C and 1000-1300 r/min for 4-5 h. Cool naturally to room temperature and stand for 24-36 h. Filter and wash, cool and dry to obtain a fishery regulating composition with lipid-lowering and blood sugar-control properties.

6. The method for preparing a fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to claim 5, characterized in that the mass ratio of cotton stalks to corn stalks in step S1 is (1-2.5): (3-5.4), the mass ratio of Aspergillus oryzae to Bacillus amyloliquefaciens in the mixed bacterial liquid is (2.5-3): (1-2.5); the mass ratio of the mixture to the volume ratio of Mandel salt solution is (1-2) g: (2-5) mL.

7. The method for preparing a fishery regulating composition with lipid-lowering and blood sugar-controlling functions according to claim 5, characterized in that the temperature of the berberine, allicin, benfotiamine, γ-aminobutyric acid, sodium oxalate and γ-glutamyl-phenylalanine when crushed in step S2 is less than 40°C.

8. The method for preparing a fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to claim 5 is characterized in that the dispersant in step S3 is a mixture of calcium lignin sulfonate and polyvinyl pyrrolidone K17 in a weight ratio of 2:1, and the methanol aqueous solution is a mixed solution of methanol and distilled water in a volume ratio of 1:3; the mass ratio of the β-cyclodextrin and the dispersant is 3:(3.5~5); the volume ratio of the total mass of the β-cyclodextrin and the dispersant to the methanol aqueous solution is (400~500)g:(1~2.5)L; the volume ratio of the methanol aqueous solution to the methanol solution of the coarse regulating and controlling composition is 20:(1~1.5).

9. Use of the fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to any one of claims 1 to 4 in the preparation of freshwater fish breeding feed.

10. The use of the fishery regulating and controlling composition with lipid-lowering and blood sugar-controlling functions according to claim 9 in the preparation of freshwater fish breeding feed, characterized in that the addition amount of the regulating and controlling composition in the freshwater fish feed is 0.5~1.4 wt%; the freshwater fish is grass carp, bighead bream, carp, crucian carp, tilapia, channel catfish, largemouth bass or black snakehead.