CN113142394A

CN113142394A - Feed additive, feed and preparation method and application thereof

Info

Publication number: CN113142394A
Application number: CN202110276229.8A
Authority: CN
Inventors: 张运强; 柳高峰; 周探科
Original assignee: Hunan Canzoho Biological Technology Co ltd
Current assignee: Hunan Canzoho Biological Technology Co ltd
Priority date: 2021-03-15
Filing date: 2021-03-15
Publication date: 2021-07-23

Abstract

The application provides a feed additive, a feed, and a preparation method and application thereof. The feed additive comprises the following components in parts by weight: 6-9 parts of schizochytrium limacinum oil, 1-4 parts of mycobacterium phlei, 1-3 parts of anthocyanin and 1-4 parts of choline chloride. Feed, including basal feed and feed additives. The preparation method of the feed comprises the following steps: mixing the basic feed with feed additive. The application of the feed is used for feeding hens. The feed additive provided by the application extracts DHA oil from schizochytrium limacinum, strengthens the absorption and conversion of DHA in a chicken body by using mycobacterium phlei and choline chloride, utilizes anthocyanin to be efficient and antioxidant, and finally achieves the purpose of efficiently and highly-pure enriching DHA in eggs.

Description

Feed additive, feed and preparation method and application thereof

Technical Field

The application relates to the field of feeds, in particular to a feed additive, a feed, and a preparation method and application thereof.

Background

The DHA eggs are eggs rich in DHA, and the chemical name of the DHA is docosahexaenoic acid, which is a polyunsaturated fatty acid very important for human bodies. DHA is a main element for the growth and maintenance of nervous system cells, is an important component of brain and retina, and has three main functions: (1) promoting brain neural development; (2) promoting development of optic nerve; (3) improving immunity. As the channel for supplementing DHA is single, a large amount of DHA can be obtained only by eating seafood products, and the fish oil capsule products have low digestibility, the DHA is enriched into eggs, so that the channel for obtaining DHA by people is high-efficiency.

The DHA in the eggs mainly exists in the form of lecithin type DHA, and is different from artificially purified DHA and DHA in natural fish oil or algae oil, the artificially purified DHA is usually ethyl ester type DHA, namely ethanol plus DHA, and the ethanol is commonly called as alcohol, so that some people allergic to the alcohol, especially pregnant women and women in lactation period, are not suitable for eating the ethyl ester type DHA. Natural DHA is often "triglyceride-type" DHA, which has a much lower digestibility than the "lecithin-type DHA in eggs. The egg is known as "whole nutrient food", and DHA egg still contains other essential nutrients in addition to being rich in DHA: such as protein, mineral, vitamin and rich long-chain unsaturated fatty acid, break the mode of single-purity DHA supplement in the health care product. DHA is enriched in eggs, so that the daily required DHA can be supplemented more comprehensively, safely and conveniently by the public, and the DHA-enriched egg has important significance for improving the health of human beings.

Although the DHA content of a lot of omega-3 rich eggs in the market is high, due to the fact that the eggs contain a large amount of linolenic acid and EPA, the DHA purity (particularly the content proportion of DHA in omega-3 unsaturated fatty acid) is low, so that a lot of challenges exist in the process of enriching DHA in the eggs: (1) because DHA belongs to long-chain unsaturated fatty acid, the conversion efficiency of the chicken group into DHA through octadecatrienoic acid (ALA, linolenic acid) is very low, so that the DHA-related additives (such as linseed and the like) are not preferable to be fed; (2) if fish oil or linseed oil with high DHA content is fed, a certain amount of EPA (eicosapentaenoic acid) (ALA) and eicosapentaenoic acid (EPA) can be produced by the body due to the fact that the EPA is rich, and the EPA can expand blood vessels, but the problems of thrombopoiesis obstacle and the like can be caused by excessive consumption of the eggs by infants, so the eggs with high EPA content are not suitable for children and pregnant women; (3) even if the feed for the chickens does not contain ALA and EPA, the organism has very limited digestion and absorption rate of DHA; (4) in addition, because unsaturated fatty acid is easily oxidized in the body, the DHA enrichment efficiency in eggs is difficult to guarantee.

In view of this, the present application is specifically made.

Disclosure of Invention

The present application aims to provide a feed additive, a feed, a preparation method and an application thereof, so as to solve the above problems.

In order to achieve the purpose, the following technical scheme is adopted in the application:

a feed additive comprises the following components in parts by weight:

6-9 parts of schizochytrium limacinum oil, 1-4 parts of mycobacterium phlei, 1-3 parts of anthocyanin and 1-4 parts of choline chloride.

The egg yolk in the egg is a precursor of the egg yolk, during the process of enriching the egg yolk by the egg yolk, the egg yolk precursor carries DHA to be combined with a specific receptor on the egg yolk and enters the egg yolk through endocytosis, and during the physiological metabolic process, the DHA enrichment in the egg yolk is influenced by the following 4 aspects: (1) DHA needs to be highly pure in the feed: excessive other fatty acids exist and can form metabolic competition effect with DHA, so that the enrichment of DHA is influenced, and the contents of linolenic acid and EPA are controlled, so that the contents of the two in eggs can be controlled; (2) health degree of intestinal mucosa of chicken organism: the health of the intestinal mucosa is the basis for guaranteeing the digestion and absorption of DHA; (3) the integral oxidation resistance of chicken organism: as DHA belongs to long-chain unsaturated fatty acid, oxidation is easy to occur, and if the organism oxidation resistance of the chicken is not strong, DHA can be consumed in the organism, thereby reducing the DHA enrichment in the eggs; (4) metabolic capacity of liver for lipids: the DHA is enriched in the eggs, mainly through a lipoprotein metabolism path, and the DHA enrichment can be influenced by the strong and weak metabolic capability of the liver on the lipoprotein.

Optionally, in the feed additive, the amount of schizochytrium oil can be any value between 6 parts, 7 parts, 8 parts, 9 parts and 6-9 parts by weight; the amount of Mycobacterium phlei may be any of 1 part, 2 parts, 3 parts, 4 parts and 1-4 parts; the amount of anthocyanins can be any value between 1 part, 2 parts, 3 parts and 1-3 parts; the choline chloride may be used in any amount between 1 part, 2 parts, 3 parts, 4 parts, and 1-4 parts.

Preferably, the feed additive comprises the following components in parts by weight: 6 parts of schizochytrium limacinum oil, 2 parts of mycobacterium phlei, 1 part of anthocyanin and 1 part of choline chloride.

Preferably, the preparation method of the schizochytrium oil comprises the following steps:

and (3) sequentially centrifuging, drying and carrying out enzymolysis on the schizochytrium to obtain the schizochytrium oil.

Preferably, the centrifugal force of the centrifugation is 10000 Xg-12000 Xg, the temperature is 3-5 ℃, and the time is 20-40 min;

preferably, the drying comprises freeze-drying at a temperature of-50 ℃ to-40 ℃ for a period of 18-30 h.

The schizochytrium oil obtained by the method has the DHA purity as high as 40% and the EPA content is low (less than or equal to 0.4%).

Optionally, the centrifugal force of the centrifugation may be any value between 10000 xg, 11000 xg, 12000 xg and 10000 xg-12000 xg, the temperature may be any value between 3 ℃, 4 ℃, 5 ℃ and 3-5 ℃, and the time may be any value between 20min, 30min, 40min and 20-40 min; the temperature of the freeze drying can be any value between-50 ℃, 45 ℃, 40 ℃ and-50 ℃ to-40 ℃, and the time can be any value between 18h, 24h, 30h and 18-30 h.

Preferably, the enzymatic hydrolysis comprises: firstly, carrying out enzymolysis for 2-4h by using neutral protease at the temperature of 40-50 ℃ and under the condition of pH 6-7; then carrying out enzymolysis for 5-7h by using alkaline protease at 65-75 ℃ and under the condition of pH9-10, and then centrifuging and layering to remove upper-layer grease.

Optionally, the temperature for enzymolysis by neutral protease can be any value between 40 ℃, 45 ℃, 50 ℃ and 40-50 ℃, the pH can be any value between 6, 6.5, 7 and 6-7, and the time can be any value between 2h, 3h, 4h and 2-4 h; the temperature for enzymolysis by alkaline protease can be 65 ℃, 70 ℃, 75 ℃ and any value between 65 ℃ and 75 ℃, the pH can be 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10 and any value between 9 and 10, and the time can be any value between 5h, 6h, 7h and 5 to 7 h.

Preferably, the mycobacterium phlei species is f.u.36;

preferably, the Mycobacterium phlei is inactivated for 15-25min at 110-130 ℃ prior to use.

The main effect of inactivation is to reduce the risk of adverse reactions caused by live bacteria.

Optionally, the temperature for inactivating Mycobacterium phlei may be any value between 110 ℃, 120 ℃, 130 ℃ and 110-.

The application also provides a feed, which comprises a basic feed and the feed additive.

Preferably, the feed additive accounts for 0.5-0.7% of the total mass of the feed.

Alternatively, the proportion of the feed additive to the total mass of the feed may be any value between 0.5%, 0.6%, 0.7% and 0.5-0.7%.

Preferably, the basal feed comprises one or more of corn, soybean meal and premix.

The premix referred to in this application may be any commonly used feed additive that is premixed. The following premix is preferably used: protein is taken as a carrier, and each kilogram of the protein provides the following contents of minerals and vitamins, namely 150mg of Fe, Cu: 20mg, Zn 150mg, Mn 30mg, Se 0.4mg, I0.3 mg, VA 17800IU, VD 35000IU, VE 44IU, VK 5mg, VB 18mg, VB2 15mg, VB6 8.5mg, VB12 0.05mg, VB3 60mg, VB5 25mg and VB9 2.5 mg.

The application also provides a preparation method of the feed, which comprises the following steps:

mixing the basal feed and the feed additive.

The application also provides an application of the feed for feeding hens.

Compared with the prior art, the beneficial effect of this application includes:

the feed additive and the feed provided by the application realize high-purity DHA purification by extracting DHA oil from schizochytrium limacinum, and the mycobacterium phlei is used for improving the digestive absorption rate of the intestinal tract (improving the nonspecific immunity of the intestinal tract mucous membrane, thereby ensuring the health of the intestinal tract mucous membrane and further achieving the purpose of efficiently absorbing DHA, if the health of the intestinal tract cannot be ensured, the absorption and the utilization of any nutrient elements cannot be mentioned at all), choline chloride is used for strengthening lipid metabolism, anthocyanin is used for efficiently resisting oxidation, and finally the purpose of efficiently and highly purifying enriched DHA of eggs is achieved, under the condition of improving the DHA content in eggs, the ALA and EPA contents are very low, the problem that alcohol allergy people generated by ethyl ester type DHA are not applicable is avoided, the egg is suitable for pregnant women, lactating women and children of low age to eat, and the egg has important significance on the development of cranial nerves and the development of optic nerves of fetuses and infants;

the preparation method of the feed is simple in process.

Drawings

To more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are required to be used in the embodiments are briefly described below, and it should be understood that the following drawings only illustrate some embodiments of the present application and therefore should not be considered as limiting the scope of the present application.

FIG. 1 is a photograph of Schizochytrium oil obtained in example 1;

FIG. 2 is a photograph of the feed obtained in example 1;

figure 3 is a photograph of feeding with the feed provided herein.

Detailed Description

The terms as used herein:

"prepared from … …" is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

The conjunction "consisting of … …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of … …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.

When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when the range "1 ~ 5" is disclosed, the ranges described should be construed to include the ranges "1 ~ 4", "1 ~ 3", "1 ~ 2 and 4 ~ 5", "1 ~ 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.

In these examples, the parts and percentages are by mass unless otherwise indicated.

"part by mass" means a basic unit of measure indicating a mass ratio of a plurality of components, and 1 part may represent any unit mass, for example, 1g or 2.689 g. If we say that the part by mass of the component A is a part by mass and the part by mass of the component B is B part by mass, the ratio of the part by mass of the component A to the part by mass of the component B is a: b. alternatively, the mass of the A component is aK and the mass of the B component is bK (K is an arbitrary number, and represents a multiple factor). It is unmistakable that, unlike the parts by mass, the sum of the parts by mass of all the components is not limited to 100 parts.

"and/or" is used to indicate that one or both of the illustrated conditions may occur, e.g., a and/or B includes (a and B) and (a or B).

Embodiments of the present application will be described in detail below with reference to specific examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present application and should not be construed as limiting the scope of the present application. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

Example 1

The embodiment provides a feed additive, which comprises the following components in parts by weight: 6 parts of schizochytrium limacinum oil, 2 parts of mycobacterium phlei, 1 part of anthocyanin and 1 part of choline chloride.

Wherein, the schizochytrium oil is prepared by fresh schizochytrium through low-speed centrifugation, freeze drying and two-step enzymolysis method in turn, and the detailed steps are as follows: (1) centrifuging at 11200 Xg centrifugal force and 4 deg.C for 30min with a low speed centrifuge; (2) removing supernatant after centrifugation, freeze-drying the residual algae mud for 24h at-45 ℃, and crushing to form dry powder; (3) performing two-step enzymolysis treatment, wherein in the first step, 7% of neutral protease is added, the pH value is adjusted to pH 6.5, the enzymolysis time is 3 hours at the temperature of 45 ℃, in the second step, 10% of alkaline protease is added, the pH value is adjusted to pH 9.4, and the enzymolysis time is 6 hours at the temperature of 68 ℃; (4) and (3) taking out the enzymatic hydrolysate after the enzymolysis is finished, centrifuging for 10min at 5000rpm, layering, and taking out the upper layer of grease, namely the purified schizochytrium oil (as shown in figure 1). The oil content of the schizochytrium limacinum oil is more than or equal to 90 percent and the DHA content is more than or equal to 40 percent. The strain of Mycobacterium phlei is F.U.36, and is inactivated at 120 deg.C for 20min before use.

The laying period of the laying hens is based on the basic feed, and the feed additive is added according to 0.6% of the total weight of the feed to obtain the feed (shown in figure 2), and the feed is mixed and fed (shown in figure 3). The obtained egg has an octadecatrienoic acid (ALA) content of less than or equal to 40mg/100g, an eicosapentaenoic acid (EPA) content of less than or equal to 15mg/100g, and a docosahexaenoic acid (DHA) content of greater than or equal to 150mg/100 g.

Example 2

The embodiment provides a feed additive, which comprises the following components in parts by weight: 6.5 parts of schizochytrium limacinum oil, 3 parts of mycobacterium phlei, 2 parts of anthocyanin and 3 parts of choline chloride.

Wherein, the schizochytrium oil is prepared by fresh schizochytrium through low-speed centrifugation, freeze drying and two-step enzymolysis method in turn, and the detailed steps are as follows: (1) centrifuging at a centrifugal force of 10000 Xg and a temperature of 3 ℃ for 40min by using a low-speed centrifuge; (2) removing supernatant after centrifugation, freeze-drying the residual algae mud at-50 ℃ for 30h, and crushing to form dry powder; (3) performing two-step enzymolysis treatment, wherein in the first step, neutral protease is added by 7%, the pH value is adjusted to pH6, and the enzymolysis is performed for 4 hours at the temperature of 40 ℃, and in the second step, alkaline protease is added by 10%, the pH value is adjusted to pH9, and the enzymolysis is performed for 7 hours at the temperature of 65 ℃; (4) and taking out the enzymatic hydrolysate after the enzymolysis is finished, centrifuging for 10min at 5000rpm, layering, and taking out the upper layer of grease, namely the purified schizochytrium oil. The oil content of the schizochytrium limacinum oil is more than or equal to 90 percent and the DHA content is more than or equal to 40 percent. The strain of Mycobacterium phlei is F.U.36, and is inactivated at 110 deg.C for 25min before use.

The feed additive is added into the laying hens in the laying period according to 0.5% of the total weight of the feed on the basis of the basic feed, and the laying hens are mixed and fed.

Example 3

The embodiment provides a feed additive, which comprises the following components in parts by weight: 9 parts of schizochytrium limacinum oil, 4 parts of mycobacterium phlei, 3 parts of anthocyanin and 4 parts of choline chloride.

Wherein, the schizochytrium oil is prepared by fresh schizochytrium through low-speed centrifugation, freeze drying and two-step enzymolysis method in turn, and the detailed steps are as follows: (1) centrifuging at 12000 Xg centrifugal force and 5 deg.C for 20min with low speed centrifuge; (2) removing supernatant after centrifugation, freeze-drying the residual algae mud at-40 deg.C for 18h, and pulverizing to obtain dry powder; (3) performing two-step enzymolysis treatment, namely adding 7% of neutral protease in the first step, adjusting the pH value to pH 7, performing enzymolysis for 3 hours at the temperature of 50 ℃, adding 10% of alkaline protease in the second step, adjusting the pH value to pH10, and performing enzymolysis for 5 hours at the temperature of 75 ℃; (4) and taking out the enzymatic hydrolysate after the enzymolysis is finished, centrifuging for 10min at 5000rpm, layering, and taking out the upper layer of grease, namely the purified schizochytrium oil. The oil content of the schizochytrium limacinum oil is more than or equal to 90 percent and the DHA content is more than or equal to 40 percent. Mycobacterium phlei strain is F.U.36, and is inactivated at 130 deg.C for 15min before use.

The feed additive is added into the laying hens in the laying period according to 0.7% of the total weight of the feed on the basis of the basic feed, and the laying hens are mixed and fed.

Comparative example 1

The basal feed was recorded as group C1 without any feed additives as a blank control.

Comparative example 2

1.2 wt% of tuna oil (product number: CAS:8016-13-5) was added to the basal feed as a feed (DHA content in the feed was 0.15%), and the feed was designated as group C2.

Comparative example 3

2 wt% of linseed oil (product number: CAS:8001-26-1) was added to the basal feed as a feed, and recorded as group C3.

Comparative example 4

0.3 wt% of schizochytrium limacinum oil is added on the basis of the basic feed to be used as feed, and the feed is recorded as T1 group.

Comparative example 5

0.3 wt% of schizochytrium limacinum oil, 0.15 wt% of mycobacterium phlei and 0.1 wt% of choline chloride are added on the basis of the basic feed, and the feed is marked as T2 group.

Comparative example 6

0.3 wt% of schizochytrium limacinum oil, 0.05 wt% of anthocyanin and 0.1 wt% of choline chloride are added on the basis of the basic feed, and the feed is marked as T3 group.

Comparative example 7

0.3 wt% of schizochytrium limacinum oil, 0.15 wt% of mycobacterium phlei and 0.05 wt% of anthocyanin are added on the basis of the basic feed, and the mixture is marked as T4 group.

Test examples

0.3 wt% of schizochytrium limacinum oil, 0.15 wt% of mycobacterium phlei, 0.05 wt% of anthocyanin and 0.1 wt% of choline chloride are added on the basis of the basic feed, and the feed is marked as T5 group.

Measurement indexes are as follows: removing shell of ovum gallus Domesticus, mixing, and detecting omega-3 unsaturated fatty acid (alpha linolenic acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)) with index of 2 g.

Summary of omega-3 unsaturated fatty acid detection:

the detection method is the gas chromatography for detecting omega-3 polyunsaturated fatty acid in NY/T2068-2011 eggs and egg products by using a gas chromatograph. Brief introduction of the method: hydrolyzing a sample with hydrochloric acid, extracting fat with diethyl ether-petroleum ether, saponifying with potassium hydroxide and methanol, performing methyl esterification with a boron trifluoride-methanol solution to obtain fatty acid methyl ester, separating by a gas chromatographic column, detecting by a hydrogen flame ionization detector, quantifying the peak area of the peak appearance time of each fatty acid by an edible internal standard method, and calculating the content of each component according to a formula.

The results are shown in table 1:

TABLE 1 test results

As shown in table 1, the results show that: (1) the DHA content of common eggs is low, and is only 25.79mg/100 g; (2) the DHA content of the eggs fed with the fish oil is greatly increased, but the EPA content of the eggs is also increased, and the DHA content of the eggs reaches 34.59mg/100 g; (3) the feed is added with 2.00 percent of linseed oil, which can well enrich DHA, so that the DHA content reaches 152.18mg/100g, but the alpha linolenic acid content reaches 388.32mg/100g, thereby greatly reducing the DHA purity and having great influence on the flavor and the nutritional structure of eggs; (4) the DHA content of two combined groups (T2, T3 and T4) which respectively use algae oil to cooperate with mycobacterium phlei, anthocyanin or choline is improved compared with that of algae oil T1, but is slightly lower than that of the group T5; (5) the DHA content of the eggs in the algae oil combination scheme group (0.3% of schizochytrium oil, 0.15% of mycobacterium phlei, 0.05% of anthocyanin and 0.05% of choline chloride) reaches 189.07mg/100g, the DHA content and purity of the eggs are greatly improved compared with those of common eggs, and the DHA content and purity of the eggs are obviously improved compared with those of the rest control group and the treatment group. In conclusion, the algae oil feed additive provided by the invention can be used for efficiently enriching DHA in eggs, and the purity of DHA in the enriched omega-3 unsaturated fatty acid is improved.

Finally, it should be noted that: the above embodiments are only used for illustrating the technical solutions of the present application, and not for limiting the same; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present application.

Moreover, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the application and form different embodiments. For example, in the claims above, any of the claimed embodiments may be used in any combination. The information disclosed in this background section is only for enhancement of understanding of the general background of the application and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.

Claims

1. The feed additive is characterized by comprising the following components in parts by weight:

2. The feed additive according to claim 1, comprising, in parts by weight: 6 parts of schizochytrium limacinum oil, 2 parts of mycobacterium phlei, 1 part of anthocyanin and 1 part of choline chloride.

3. The feed additive of claim 1, wherein the schizochytrium oil is prepared by a method comprising:

4. The feed additive according to claim 3, wherein the centrifugal force of the centrifugation is 10000 Xg-12000 Xg, the temperature is 3-5 ℃, and the time is 20-40 min;

5. The feed additive of claim 3, wherein the enzymatic hydrolysis comprises: firstly, carrying out enzymolysis for 2-4h by using neutral protease at the temperature of 40-50 ℃ and under the condition of pH 6-7; then carrying out enzymolysis for 5-7h by using alkaline protease at 65-75 ℃ and under the condition of pH9-10, and then centrifuging and layering to remove upper-layer grease.

6. The feed additive of any one of claims 1-5 wherein the Mycobacterium phlei species is F.U.36;

7. A feed comprising a basal feed and a feed additive according to any one of claims 1 to 6.

8. The feed of claim 7, wherein the feed additive comprises 0.5% to 0.7% of the total mass of the feed;

9. A method of preparing the feed of claim 7 or 8, comprising:

mixing the basal feed and the feed additive.

10. Use of the feed according to claim 7 or 8 for feeding hens.