CN110651917A

CN110651917A - Feed additive for protecting fish liver under oxidative stress condition and preparation method thereof

Info

Publication number: CN110651917A
Application number: CN201911074254.7A
Authority: CN
Inventors: 王杰; 吉中力; 陈春明
Original assignee: WUXI HANOVE ANIMAL HEALTH PRODUCTS CO Ltd
Current assignee: WUXI HANOVE ANIMAL HEALTH PRODUCTS CO Ltd
Priority date: 2019-11-06
Filing date: 2019-11-06
Publication date: 2020-01-07

Abstract

The invention discloses a feed additive for protecting fish liver under oxidative stress and a preparation method thereof, wherein the feed additive comprises the following raw materials in percentage by mass: 2-8% of tea polyphenol, 8-15% of selenium, 20-28% of vitamin C, 20-25% of vitamin E, 15-20% of liquorice, 10-15% of polygonum cuspidatum, 5-10% of lycium barbarum polysaccharide and the balance of filler. According to the feed additive for protecting the liver of the fish under the oxidative stress condition and the preparation method thereof, tea polyphenol can play a synergistic role with selenium, vitamin C, vitamin E, liquorice, giant knotweed and other Chinese herbal medicines, active oxygen free radicals can be eliminated, the oxidation resistance of an organism can be improved, free radical disorder caused by oxidation of feed grease can be regulated, energy metabolism is promoted, damaged hepatic cells are repaired, the liver of the fish can be protected under the oxidative stress condition, and the healthy growth of the fish can be promoted.

Description

Feed additive for protecting fish liver under oxidative stress condition and preparation method thereof

Technical Field

The invention belongs to the technical field of feed additives, and particularly relates to a feed additive for protecting fish livers under oxidative stress conditions and a preparation method thereof.

Background

As one of the important essential nutrients for animals, fats and oils have various physiological functions in the living metabolism of the body, such as: can provide energy for organism, participate in tissue cell constitution, promote absorption of fat-soluble vitamins, provide essential fatty acid for organism, etc. In recent years, as the prices of high-quality animal proteins such as fish meal and the like are gradually increased, various large feed enterprises try to improve the oil content in the feed to stimulate the energy supply efficiency of the fish to fat, so as to reduce the dependence on the fish meal and achieve the purpose of saving the cost.

Long-term cultivation practice shows that the improvement of the oil content in the feed has certain promotion effect on the growth of cultivated fishes. However, since the oil contains a large amount of unsaturated bonds, the oil is easily oxidized and rancid by high temperature and high humidity during the transportation, storage and processing of the feed, and various aldehydes, ketones, acids and hydrocarbon compounds are generated. When the intake of these lipid oxidation products exceeds the detoxification capability of the liver, liver cells are damaged, inflammatory mediators and carcinogenic factors are released, and hepatic fibrosis and hepatobiliary syndromes such as fatty liver are caused in severe cases.

In view of the many adverse consequences of fat oxidation, most feeds will have antioxidants added to them. The commonly used antioxidants at present comprise dibutyl p-cresol (BHT), Butyl Hydroxyanisole (BHA), tert-butyl hydroquinone (TBHQ) and the like, but the antioxidants are chemical synthesis products, have low toxicity and have certain potential safety hazards, for example, the butyl hydroxyanisole may cause liver injury, even liver cancer and the like. Some natural antioxidants have the problems of high price, poor stability and the like. Therefore, starting from the cultured fish, how to solve the problem of oxidation of feed grease is important, and the improvement of the oxidation resistance and the detoxification capability of the liver is important.

Disclosure of Invention

In order to solve the technical problems in the prior art, the invention aims to provide the feed additive for protecting the liver of the fish under the condition of oxidative stress and the preparation method thereof.

In order to achieve the purpose and achieve the technical effect, the invention adopts the technical scheme that:

the feed additive for protecting the liver of the fish under the oxidative stress condition comprises the following raw materials in percentage by mass: 2-8% of tea polyphenol, 8-15% of selenium, 20-28% of vitamin C, 20-25% of vitamin E, 15-20% of liquorice, 10-15% of polygonum cuspidatum, 5-10% of lycium barbarum polysaccharide and the balance of filler.

Further, the selenium is selenium yeast.

Further, the filler is talcum powder.

The invention also provides a preparation method of the feed additive for protecting the liver of the fish under the oxidative stress condition, which comprises the following steps:

step (1), weighing a proper amount of tea polyphenol, selenium, vitamin C, vitamin E, liquorice, polygonum cuspidatum, lycium barbarum polysaccharide and fillers according to the mass percentage of 100% for later use;

step (2), mixing the liquorice and the giant knotweed rhizome weighed in the step (1), carrying out superfine grinding, and sieving the micro powder with a 300-400-mesh sieve to obtain a micro powder mixture;

step (3), the micro powder mixture prepared in the step (2) is cooked for 4-6 hours at the temperature of 80-100 ℃, and then is filtered to obtain a cooking filtrate;

step (4), fully mixing the tea polyphenol, selenium, vitamin C, vitamin E and lycium barbarum polysaccharide weighed in the step (1) to obtain a mixture, and then directly and uniformly spraying the cooking filtrate prepared in the step (3) on the surface of the mixture;

and (5) uniformly mixing the mixture of the sprayed filtrate obtained in the step (4) with fillers to obtain the required feed additive.

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

(1) the tea polyphenol can remove redundant free radicals or peroxides in fish bodies by supplying electron hydrogen, improve the activity of main antioxidant enzymes of organisms such as GPx, SOD, CAT and the like, can regenerate vitamin E, directly protect and repair an antioxidant system, and further improve the antioxidant capacity of the organisms; a plurality of oxidation reactions in a machine body need catalysis of metal ions, the tea polyphenol used by the invention can complex a plurality of metal ions, not only can reduce the oxidation reaction rate, but also has the detoxification function, and in addition, the tea polyphenol can also play a synergistic effect with other antioxidant substances, such as selenium, vitamin C, vitamin E and the like;

(2) selenium (Se) is an important component of glutathione peroxidase (GSH-Px), and can protect the body from oxidative damage of peroxidation products and free radicals to cells, so that the integrity and functionality of the cell structure are maintained;

(3) vitamin C is one of essential micronutrients for maintaining normal physiological functions of fishes, and can also be used as an antioxidant and an immunopotentiator, wherein the vitamin C can promote cell growth, eliminate excessive free radicals in cells, increase the activity of glutathione peroxidase in the cells and improve the oxidation stress resistance of organisms;

(4) vitamin E is one of essential nutrients of fish, has the functions of maintaining fish quality, improving immunity, preventing cell hemolysis and the like, and is used as a fat-soluble antioxidant to participate in oxygen radical removal in fish bodies to terminate lipid peroxidation;

(5) the lycium barbarum polysaccharide can relieve mitochondrial injury and remove hydroxyl free radicals;

(6) the liquorice is the king of a popular medicine, has the efficacy of removing the toxin, and the extract of the liquorice has the functions of resisting inflammation and virus, protecting the liver, detoxifying, enhancing immunity and the like;

(7) the giant knotweed rhizome extract has the effects of promoting microcirculation of damaged liver tissues, promoting regeneration and repair of liver cells and inhibiting adhesion of white blood cells, platelets and liver endothelial cells;

(8) the Chinese herbal medicines (liquorice and giant knotweed) are superfined to break cell walls, so that effective ingredients are efficiently released in the cooking process, and the efficacy is improved;

(9) the product is a mixture of Chinese herbal medicine extracts, tea polyphenol, selenium, vitamin C, vitamin E and lycium barbarum polysaccharide, the finished product is powder, and compared with the traditional liver-protecting and anti-stress powder, the product has the advantages of quick response, strong efficacy and low addition amount, and can take effect only by 0.10-0.20%;

(10) the invention is suitable for various aquaculture fishes, is particularly suitable for cultivating fishes which are high in density and easy to stress, and can effectively reduce liver damage caused by stress such as grease oxidation and the like after long-term use, protect liver health and promote healthy growth of the fishes.

Drawings

FIG. 1a is a liver anatomical map of a grass carp of a second oxidized fat control group in example 7 of the present invention;

FIG. 1b is a liver anatomical diagram of grass carp of the test group in example 7 of the present invention;

fig. 2a is a liver anatomical diagram of a crucian carp before feeding a feed additive in example 9 of the present invention;

fig. 2b is a liver anatomical diagram of crucian carp fed with the feed additive in example 9 of the present invention.

Detailed Description

The embodiments of the present invention will be described in detail with reference to the accompanying drawings so that the advantages and features of the invention can be more easily understood by those skilled in the art, and the scope of the invention will be clearly and clearly defined.

Example 1

The preparation method of the feed additive for protecting the liver of the fish under the oxidative stress condition comprises the following steps:

step (1), weighing 2% of tea polyphenol, 10% of selenium yeast, 20% of vitamin C, 20% of vitamin E, 18% of liquorice, 13% of giant knotweed, 8% of lycium barbarum polysaccharide and 9% of talcum powder according to 100% of mass percent.

And (2) mixing the liquorice and the giant knotweed rhizome weighed in the step (1), carrying out superfine grinding, and sieving micro powder with a 300-mesh sieve to obtain a micro powder mixture.

And (3) cooking the micro powder mixture at 100 ℃ for 4 hours, and filtering to obtain a cooking filtrate.

And (4) fully mixing the tea polyphenol, selenium, vitamin C, vitamin E and lycium barbarum polysaccharide weighed in the step (1) to obtain a mixture, and then directly and uniformly spraying the cooking filtrate obtained in the step (3) on the surface of the mixture.

And (5) uniformly mixing the mixture sprayed with the cooking filtrate obtained in the step (4) with talcum powder to obtain the feed additive for protecting the liver health of the fish.

Example 2

step (1), weighing 4% of tea polyphenol, 8% of selenium yeast, 22% of vitamin C, 20% of vitamin E, 20% of liquorice, 15% of giant knotweed, 5% of lycium barbarum polysaccharide and 6% of talcum powder according to 100% of mass percent.

And (2) mixing the liquorice and the giant knotweed rhizome weighed in the step (1), carrying out superfine grinding, and sieving by a 300-mesh sieve to obtain a micro powder mixture.

And (3) cooking the micro powder mixture at 80 ℃ for 6 hours, and filtering to obtain a cooking filtrate.

Example 3

step (1), weighing 6% of tea polyphenol, 12% of selenium yeast, 20% of vitamin C, 20% of vitamin E, 15% of liquorice, 10% of giant knotweed, 10% of lycium barbarum polysaccharide and 7% of talcum powder according to 100% of mass percent.

And (3) cooking the micro powder mixture at 100 ℃ for 5 hours, and filtering to obtain a cooking filtrate.

And (4) fully mixing the tea polyphenol, the selenium, the vitamin C, the vitamin E and the lycium barbarum polysaccharide to obtain a mixture, and then directly and uniformly spraying the obtained cooking filtrate on the surface of the mixture.

Example 4

step (1), weighing 8% of tea polyphenol, 15% of yeast selenium, 20% of vitamin C, 20% of vitamin E, 15% of liquorice, 10% of giant knotweed, 5% of wolfberry polysaccharide and 7% of talcum powder according to 100% of mass percent.

And (3) cooking the micro powder mixture at 100 ℃ for 6 hours, and filtering to obtain a cooking filtrate.

Example 5

step (1), weighing 6% of tea polyphenol, 10% of selenium yeast, 28% of vitamin C, 22% of vitamin E, 15% of liquorice, 10% of giant knotweed, 5% of lycium barbarum polysaccharide and 4% of talcum powder according to 100% of mass percent.

And (4) fully mixing the tea polyphenol, selenium, vitamin C, vitamin E and lycium barbarum polysaccharide weighed in the step (1) to obtain a mixture, and then directly and uniformly spraying the obtained cooking filtrate on the surface of the mixture.

Example 6

step (1), weighing 4% of tea polyphenol, 10% of selenium yeast, 25% of vitamin C, 25% of vitamin E, 15% of liquorice, 10% of giant knotweed, 5% of lycium barbarum polysaccharide and 6% of talcum powder according to 100% of mass percent.

And (3) cooking the micro powder mixture at 90 ℃ for 6 hours, and filtering to obtain a cooking filtrate.

Example 7

Selecting 600 healthy grass carp tails, weighing 480g +/-10 g initially, randomly dividing into 3 treatment groups, namely a control group I, an oxidized grease control group II and a test group, repeating 4 treatment groups for each treatment group, repeating 50 grass carp tails for each treatment group, and culturing for 3 months in 2m 1m pond net cages. The nutrient water of the feed of the 3 treatment groups is 28% of protein and 7% of fat on average, wherein the feed formula of the first control group is fish feed containing fresh soybean oil, the preparation method of the feed of the second oxidized fat control group is that the fresh soybean oil in the feed formula of the first control group is completely replaced by the oxidized soybean oil, the feed additive provided by the invention is not added to the first control group and the second oxidized fat control group, and the preparation method of the feed in the test group is that the feed additive prepared in the example 1 is added on the basis of the feed of the second oxidized fat control group in a mass ratio of 0.1%. Table 1-1 shows the influence of feed additives for protecting the livers of fishes on the growth of grass carps, the survival rate, the weight gain rate and the feed coefficient of grass carps are shown in Table 1-1, and the liver health conditions of grass carps in the oxidized fat control group II and the test group II are shown in figure 1a and figure 1 b.

TABLE 1-1

	Control group one	Control group II of oxidized oil	Test group
				Survival rate (%)	92±4^a	78±2^c	86±2^b
Weight gain (%)	316.8±10.2^a	220.4±8.6^c	280.2±4.2^ab
				Coefficient of feed	1.35±0.1^c	2.01±0.2^a	1.68±0.2^b

Note: in Table 1-1, the different lower case letters on the shoulder marks of the same row indicate significant differences.

Tables 1-2 show the effect of the feed additive for protecting the liver of fish on the liver ratio, antioxidant capacity and injury repair of grass carp, wherein the liver ratio, glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT), Malondialdehyde (MDA) and glutathione peroxidase (GSH-PX) are shown in tables 1-2.

Tables 1 to 2

Note: in tables 1-2, the different lower case letters on the shoulder marks of the same row indicate significant differences.

The test result shows that compared with the control group I, the liver is damaged by the oxidized oil, the growth performance of the grass carp in the oxidized oil control group II is obviously reduced, the liver is obviously enlarged, the oxidation resistance is obviously reduced, the color of the liver is whitish, and the damage is serious, as shown in figure 1 a; the experimental group added the feed additive prepared in the embodiment 1 into the fish feed containing oxidized soybean oil according to the mass ratio of 0.1% and fed the fish feed, so that the survival rate of the grass carp is remarkably improved, the weight gain rate of the grass carp is remarkably increased, enzyme activity indexes (AST and ALT) reflecting liver injury are remarkably reduced, the content of malondialdehyde is remarkably reduced, the antioxidant enzyme GSH-PX is remarkably increased, the color of the liver is ruddy, and the health condition is remarkably improved, as shown in figure 1 b. The results show that the feed additive can effectively improve the growth performance of the grass carp, enhance the liver oxidation resistance of the grass carp, relieve the oxidative stress damage of grease, and repair damaged liver cells.

Example 8

Selecting 360-tailed pelteobagrus fulvidraco fish with healthy physique, weighing 16.2 +/-0.4 g initially, randomly dividing the pelteobagrus fulvidraco fish into 4 treatment groups, namely a control group, an oxidized fat control group, a test group I and a test group II, repeating the treatment groups for 3 times, and repeating the treatment groups for 30 fish for 2 months in an indoor circulating water system. The nutrient water of the feed of the 4 treatment groups is 38% of protein and 10% of fat on average, wherein the feed formula of the control group is fish feed containing fresh soybean oil, the preparation method of the feed in the oxidized fat control group is that the fresh soybean oil in the feed formula of the control group is completely replaced by the oxidized soybean oil, the feed additive disclosed by the invention is not added in the control group and the oxidized fat control group, and the feed additive prepared in the example 2 is respectively added in the first test group and the second test group on the basis of the feed of the oxidized fat control group according to the mass ratio of 0.1% and 0.2%. The experiment is divided into two stages in total, including: the method comprises a liver oxidative stress injury molding stage, namely inducing the liver injury of the pelteobagrus fulvidraco by using a feed of an oil oxidation control group, and further comprises a stage of adding a feed additive for protecting the liver health of the fish to treat the liver injury, namely feeding the pelteobagrus fulvidraco with the liver injury after the liver injury of the fish, wherein molding results are shown in a table 2-1, the table 2-1 shows the influence of the oxidized oil on the liver health of the pelteobagrus fulvidraco, and the treatment effect of the feed additive for protecting the liver of the fish on the liver injury repair is shown in a table 2-2.

TABLE 2-1

	Control group	Control group of oxidized oil
			Liver body ratio (%)	1.2±0.1	1.5±0.2
Liver crude fat (%, dry weight)	19.2±0.2^a	30.5±0.3^b
			AST(U/L)	108.8±0.3^a	234.1±0.5^b
ALT(U/L)	1.8±0.2^a	7.6±0.1^b

Note: in Table 2-1, the different lower case letters on the shoulder marks of the same row indicate significant differences.

Tables 2 to 2

Note: the shoulder marks of the same row have different lower case letters to show remarkable difference.

The results show that the damage of the fat oxidation to the liver is particularly obvious for the pelteobagrus fulvidraco which is mainly carnivorous fish, and fat metabolism abnormality caused by liver cell damage can often cause fat deposition in the liver and induce liver and gall syndrome such as fatty liver and the like. In the experiment, after the liver injury of the pelteobagrus fulvidraco is induced, the experimental material of the feed additive prepared in the embodiment 2 with the addition amount of 0.1% and 0.2% is respectively used for continuously feeding the pelteobagrus fulvidraco with the liver injury. According to experimental results, after the feed additive is used, the damage to the liver of the pelteobagrus fulvidraco is obviously repaired, the AST and ALT enzyme activities are obviously reduced, the oxidation resistance of the liver is obviously enhanced, the MDA is obviously reduced, the GSH-Px is obviously increased, the fat metabolism of the liver is recovered to be normal, and the fat content of the liver is obviously reduced.

Example 9

The Changzhou gold jar farmer has the pond mouth with the scale of 20 mu, is used for intensive culture of crucian, and has the stocking density of 2000 fish/mu and the average specification of 13 fish/jin. In 8-month middle of 2019, the liver of crucian is found to turn white and have obvious pathological changes, as shown in fig. 2 a. The reason is that in summer, the high temperature and rain in the golden jar city, the ventilation of the storehouse for storing the crucian puffing materials is poor, the humidity is high, some open feeds can obviously smell a rancid smell, and the liver damage of the crucian caused by the oxidation of grease generated in the crucian puffing materials is suspected. In late 8 months in 2019, the farmer started using the crucian carp puffed feed added with 0.5% of the feed additive prepared in the embodiment 3, and the crucian carp is sampled after 20 days, so that the liver damage degree of the crucian carp is obviously improved, the liver color is ruddy, and the skin and hair are smooth, which is shown in fig. 2 b. The test results prove that the invention can obviously improve the liver damage caused by grease oxidation, enhance the anti-stress capability of cultured fishes and protect the liver health.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification and drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims

1. The feed additive for protecting the liver of the fish under the oxidative stress condition is characterized by comprising the following raw materials in percentage by mass: 2-8% of tea polyphenol, 8-15% of selenium, 20-28% of vitamin C, 20-25% of vitamin E, 15-20% of liquorice, 10-15% of polygonum cuspidatum, 5-10% of lycium barbarum polysaccharide and the balance of filler.

2. The feed supplement for protecting the liver of fish under oxidative stress as claimed in claim 1, wherein the selenium is selenium yeast.

3. The feed additive for protecting the liver of fish under oxidative stress as claimed in claim 1, wherein the filler is talc.

4. A method for preparing a feed additive for protecting the liver of fish under oxidative stress as claimed in any one of claims 1 to 3, comprising the steps of:

step (2), mixing the liquorice and the giant knotweed rhizome weighed in the step (1), carrying out superfine grinding, and sieving the micropowder with a 300-mesh and 400-mesh sieve to obtain a micropowder mixture;