A probiotic preparation for promoting livestock and poultry growth and preparation method thereof

Abstract

The present invention relates to a probiotic preparation for promoting the growth of livestock and poultry, which uses edible substances to wrap probiotics with modified chitosan with flocculation effect, so that the probiotic content in the probiotic preparation is increased, the ecological regulation effect on the water body is enhanced, the water quality stability of the water body is improved, and it is beneficial to the growth of aquatic livestock and poultry. In the preparation of the probiotic preparation, chitosan is added to have a flocculation effect on the probiotics, and the concentration of the probiotics in the preparation is increased. Moreover, the addition of single-terminated amino polyethylene glycol can introduce a large number of amino groups into the chitosan, improve the water solubility and biocompatibility of the chitosan, thereby improving the flocculation effect of the chitosan on the probiotics, thereby achieving the effect of increasing the concentration of the probiotics in the preparation.

Description

Technical Field

[0001] The invention relates to the technical field of feed additives, and in particular to a probiotic preparation for promoting the growth of livestock and poultry and a preparation method thereof. Background Art

[0002] Probiotics refer to organisms or substances that can promote the balance of intestinal microorganisms. Applying probiotics in aquaculture water or aquatic feed can improve the aquaculture ecological environment, enhance the immunity of farmed animals, inhibit pathogenic microorganisms, thereby reducing the occurrence of diseases and promoting the growth of aquatic livestock and poultry. However, in the process of livestock and poultry farming, the pH value of the aquaculture water, the content of nitrates, excess bait and feces, etc., are deposited at the bottom of the pond, the algae increase, the dissolved oxygen decreases, and its decomposition causes an increase in the content of N and P in the water. A series of problems will cause the water body to deteriorate seriously, destroy the pond ecosystem, and cause the water quality stability to be poor, which is not conducive to the growth of aquatic livestock and poultry.

[0003] The Chinese invention patent with application number CN201810187135.1 discloses a probiotic preparation for improving water quality in aquaculture and a preparation method thereof, which is prepared by mixed bacterial segmented liquid fermentation or solid-state fermentation of Lactobacillus plantarum TK9, Bifidobacterium animalis 937, Lactobacillus paracasei TK1501, Bifidobacterium adolescentis TK99, Bacillus coagulans TQ33 and Bacillus subtilis TK-1. The probiotic preparation can not only significantly improve water quality, but also the probiotic can synthesize multiple vitamins required by aquatic animals in the breeding industry, which helps to increase the body's nutrient absorption and enhance the body's ability to digest and absorb feed, but the probiotic preparation has poor stability, which is not conducive to the regulation of water bodies and the growth of livestock and poultry.

[0004] A Chinese invention patent with application number CN201510653786.1 discloses a bottom improver for aquaculture ponds. The recycled garbage particles with a special particle size after ball milling have excellent adsorption effect, which not only effectively solves the problem of construction waste disposal, but also provides a new improver adsorption body with good adsorption capacity; the probiotic mixed preparation can effectively promote the decomposition of pond bottom bacteria, thereby optimizing water quality; however, its production process is only a simple compound of several raw materials. After the construction waste is used as the bacterial agent carrier and is put into the water body, it is easy to settle to the bottom of the water, and the purification effect on the water quality is not long-lasting.

[0005] In summary, in order to solve the above-mentioned defects, it is urgently necessary to develop a probiotic preparation that promotes the growth of livestock and poultry, so as to solve the problem that the probiotic preparations in the prior art have poor stability, resulting in poor regulation ability of probiotics on water bodies, which is not conducive to the growth of aquatic livestock and poultry. Summary of the invention

[0006] Based on this, the present invention has developed a probiotic preparation for promoting the growth of livestock and poultry, which uses edible substances to encapsulate probiotics and modified chitosan with flocculation effect, thereby increasing the probiotic content in the probiotic preparation, enhancing the ecological regulating effect on water bodies, and improving the water quality stability of water bodies, which is beneficial to the growth of aquatic livestock and poultry.

[0007] One object of the present invention is to provide a probiotic preparation for promoting the growth of livestock and poultry, wherein the probiotic preparation for promoting the growth of livestock and poultry has a core-shell structure and comprises the following components in parts by weight:

[0008] Nuclear structure component A:

[0009] 30-40 servings of probiotic liquid

[0010] 50-70 copies of culture medium;

[0011] Nuclear structural component B:

[0012] Modified chitosan 10-15 parts

[0013] Alum 10-15 parts

[0014] 5-10 parts of modified hydroxylated silicon dioxide;

[0015] Shell structure:

[0016] edible substances;

[0017] The modified chitosan is a product obtained by cross-linking chitosan with glutaraldehyde and single-terminated amino polyethylene glycol;

[0018] The modified hydroxylated silicon dioxide is a product of partially grafting fatty acids onto hydroxylated silicon dioxide;

[0019] The edible substance is a viscous mixture of starch, corn flour, sweet potato flour and water.

[0020] Furthermore, the fatty acid is selected from one or more of oleic acid, linoleic acid, linolenic acid, and isostearic acid.

[0021] Furthermore, the mass ratio of the starch, corn flour, sweet potato flour and water is 4-7:3-4:4-5:8-12.

[0022] Furthermore, the probiotic liquid is a mixed bacterial liquid of Bacillus subtilis, Lactobacillus plantarum and Candida albicans.

[0023] Furthermore, the culture medium is a mixture of inorganic salts, trace elements and water.

[0024] Further, the inorganic salt is selected from one or more of nitrates, phosphates, sulfates, and carbonates;

[0025] Furthermore, the trace elements are selected from one or more of potassium salts, calcium salts, zinc salts, magnesium salts and sodium salts.

[0026] Preferably, the inorganic salt is potassium nitrate, disodium hydrogen phosphate, magnesium sulfate, and calcium carbonate.

[0027] Furthermore, the mass ratio of potassium nitrate, disodium hydrogen phosphate, magnesium sulfate, and calcium carbonate is 1.5-2.5:1.5-3:0.5-2.5:2-3.5.

[0028] Furthermore, the preparation method of the culture medium is: dissolving potassium nitrate, disodium hydrogen phosphate, magnesium sulfate, and calcium carbonate in distilled water, adding appropriate amount of growth factors and appropriate amount of trace elements, stirring evenly and adjusting the pH value to 6.5-7.0.

[0029] Furthermore, in the probiotic liquid,

[0030] The content of Bacillus subtilis is 2-5×10 8 cfu / g;

[0031] The content of plant lactobacillus is 3-6×10 8 cfu / g;

[0032] The content of Candida is 1-4×10 8 cfu / g.

[0033] The present invention also provides a method for preparing a probiotic preparation for promoting the growth of livestock and poultry, and the method for preparing the probiotic preparation for promoting the growth of livestock and poultry comprises the following steps:

[0034] S1. Preparation of modified chitosan: dissolving chitosan in acetic acid solution, stirring and adding alkali to precipitate it; then adding glutaraldehyde and single-terminated amino polyethylene glycol, and aluminum sulfate, and heating to react;

[0035] S2. Preparation of modified hydroxylated silicon dioxide: adding hydroxylated silicon dioxide into liquid fatty acid, ultrasonicating and heating;

[0036] S3, blending the probiotic solution and culture medium to obtain the core structure component A; blending the modified chitosan, modified hydroxylated silica and alum to obtain the core structure component B;

[0037] S4. Blending the core structure component A and the core structure component B, and wrapping them with the edible substance to obtain the probiotic preparation for promoting the growth of livestock and poultry.

[0038] Furthermore, in step S1, the mass ratio of chitosan, glutaraldehyde and single-terminated amino polyethylene glycol is 1:3-5:1-2.

[0039] Furthermore, in step S2, the heating temperature is 70-80°C, and the ultrasonic time is 10-30 minutes.

[0040] Furthermore, in step S1, the pH after adding alkali is 8-10.

[0041] Furthermore, the alkali is selected from one or more of sodium hydroxide and potassium hydroxide.

[0042] Furthermore, in the single-terminated amino polyethylene glycol, the number of repeating units is 8-12.

[0043] The present invention has the following beneficial effects:

[0044] 1. The present invention adds chitosan when preparing the probiotic preparation, which has a flocculation effect on the probiotics and increases the concentration of the probiotics in the preparation. Since the single-terminated amino polyethylene glycol is a molecule with a hydrophobic end and a hydrophilic center, it can be introduced into chitosan, which can increase the hydrophilicity of chitosan, thereby increasing its water solubility, and at the same time can improve the biocompatibility of chitosan, enhance the compatibility and selectivity of chitosan with probiotics, thereby improving the flocculation effect of chitosan on the probiotics, thereby achieving the effect of increasing the concentration of probiotics in the preparation.

[0045] 2. In the preparation of the probiotic preparation, the present invention adds liquid fatty acid to the hydroxylated silica, thereby improving the compatibility between the hydroxylated silica and the modified chitosan, and the three-dimensional network between the long-chain molecules on the modified hydroxylated silica and the modified chitosan and the probiotics improves the flocculation effect of chitosan on the probiotics, thereby ensuring that the probiotic preparation has the advantages of high flocculation rate, small dosage, and high probiotic content, which is beneficial to water quality regulation, thereby regulating the ammonia nitrogen content and nitrite content in the water body, making the water quality stable, and being beneficial to the growth of aquatic poultry and livestock. DETAILED DESCRIPTION

[0046] In order to more clearly illustrate the technical solution of the present invention, the following examples are listed. Unless otherwise stated, the raw materials, reactions and post-treatment methods shown in the examples are common raw materials on the market and technical methods well known to those skilled in the art.

[0047] The words "preferred", "preferably", "more preferably", etc. in the present invention refer to embodiments of the present invention that can provide certain beneficial effects in certain circumstances. However, other embodiments may also be preferred under the same circumstances or other circumstances. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the present invention.

[0048] It should be understood that, except in any operating examples, or where otherwise indicated, all numbers indicating, for example, the amounts of ingredients used in the specification and claims should be understood to be modified in all cases by the term "about". Therefore, unless indicated to the contrary, the numerical parameters set forth in the following specification and the appended claims are approximate values ​​that vary depending on the desired properties to be obtained by the present invention.

[0049] In the embodiments of the present invention, the number of parts refers to the number of parts by mass.

[0050] In an embodiment of the present invention, the probiotic liquid is a mixed liquid of Bacillus subtilis, Lactobacillus plantarum, and Candida oceani;

[0051] In the probiotic solution,

[0052] The content of Bacillus subtilis is 2-5×10 8 cfu / g;

[0053] The content of plant lactobacillus is 3-6×10 8 cfu / g;

[0054] The content of Candida is 1-4×10 8 cfu / g.

[0055] The trace elements are potassium nitrate, calcium carbonate, zinc chloride, magnesium sulfate and sodium chloride in a mass ratio of 1:1:1:1:1.

[0056] The inorganic salts are potassium nitrate, disodium hydrogen phosphate, magnesium sulfate, and calcium carbonate.

[0057] The preparation steps of the culture medium are as follows: dissolving 2 parts of potassium nitrate, 2.5 parts of disodium hydrogen phosphate, 1.5 parts of magnesium sulfate, and 3 parts of calcium carbonate in 1200 parts of distilled water, adding 0.5 parts of growth factor and 2.5 parts of trace elements, stirring evenly and adjusting the pH value to 7.0 to obtain the culture medium.

[0058] The edible substance is a viscous mixture of starch, corn flour, sweet potato flour and water; the mass ratio of the starch, corn flour, sweet potato flour and water is 5:3:4.5:10.

[0059] The preparation method of the edible substance is as follows: weigh the raw materials according to the above mass ratio, dissolve starch, corn flour and sweet potato flour in distilled water, heat to 55° C., mix evenly, and cool to room temperature to obtain the edible substance.

[0060] The single-terminated amino polyethylene glycol is selected from the amino-deca polyethylene glycol with model number SY251162 produced by Shaoyuan Company.

[0061] The chitosan in the examples of the present invention was purchased from Qingdao Honghai Biotechnology Co., Ltd., with a deacetylation degree of 91.0%.

[0062] Example 1

[0063] A probiotic preparation for promoting the growth of livestock and poultry, the probiotic preparation for promoting the growth of livestock and poultry having a core-shell structure, comprising the following components in parts by weight:

[0064] Nuclear structure component A:

[0065] Probiotic liquid 30 servings

[0066] Culture medium 50 copies;

[0067] Nuclear structural component B:

[0068] Modified chitosan 10 parts

[0069] 10 parts alum

[0070] 5 parts of modified hydroxylated silica;

[0071] Shell structure:

[0072] Edible substances; 400 portions.

[0073] The preparation method of the probiotic preparation for promoting livestock and poultry growth comprises the following steps:

[0074] S1. Preparation of modified chitosan: dissolve chitosan in an acetic acid solution with a pH of 5, stir, add sodium hydroxide solution to a pH of 8 to precipitate it, and filter it; then add glutaraldehyde and aluminum sulfate, pre-react in a water bath at 80°C for 0.5h, then add aminodecapolyethylene glycol and continue to react for 2h, precipitate it with acetone after the reaction is completed, filter it, then wash the product with ethanol, filter it, and dry it in an oven at 90°C to obtain modified chitosan; wherein the mass ratio of the chitosan, glutaraldehyde, aminodecapolyethylene glycol and aluminum sulfate is 1:3:1:1.5;

[0075] S2. Preparation of modified hydroxylated silica: add 1 part of hydroxylated silica to 10 parts of linoleic acid, perform ultrasonic treatment for 10 minutes and heat at 70°C. After the reaction is completed, place the mixture in an oven at 50°C and bake to constant weight to obtain modified hydroxylated silica;

[0076] S3, blending the above-mentioned probiotic solution and culture medium in the above-mentioned mass fractions, and culturing at 35° C. for 100 hours to obtain the core structure component A; blending the above-mentioned mass fractions of modified chitosan, modified hydroxylated silica and alum to obtain the core structure component B;

[0077] S4. Blending the core structure component A and the core structure component B in the above-mentioned mass fractions, and wrapping them with the edible substance to obtain the probiotic preparation for promoting the growth of livestock and poultry.

[0078] Example 2

[0079] A probiotic preparation for promoting the growth of livestock and poultry, the probiotic preparation for promoting the growth of livestock and poultry having a core-shell structure, comprising the following components in parts by weight:

[0080] Nuclear structure component A:

[0081] Probiotic liquid 35 servings

[0082] 60 copies of culture medium;

[0083] Nuclear structural component B:

[0084] Modified chitosan 12 parts

[0085] 12 parts alum

[0086] 7 parts of modified hydroxylated silica;

[0087] Shell structure:

[0088] Edible substances; 600 portions;

[0089] The preparation method of the probiotic preparation for promoting livestock and poultry growth comprises the following steps:

[0090] S1. Preparation of modified chitosan: dissolve chitosan in an acetic acid solution with a pH of 5, stir, and then add a sodium hydroxide solution to a pH of 9 to precipitate it; filter with suction; then add glutaraldehyde and aluminum sulfate, pre-react in a water bath at 80° C. for 1 hour, then add amino-deca-polyethylene glycol and continue to react for 3 hours, precipitate with acetone after the reaction is completed, filter with suction, then wash the product with ethanol, filter with suction, and dry in an oven at 90° C. to obtain modified chitosan; wherein the mass ratio of the chitosan, glutaraldehyde, amino-deca-polyethylene glycol and aluminum sulfate is 1:4:1.5:2.5;

[0091] S2. Preparation of modified hydroxylated silica: 2 parts of hydroxylated silica were added to 12 parts of linoleic acid, ultrasonicated for 25 minutes and heated at 75°C. After the reaction was completed, the mixture was placed in an oven at 50°C and dried to constant weight to obtain modified hydroxylated silica;

[0092] S3, blending the above-mentioned probiotic solution and culture medium in the above-mentioned mass fractions to obtain the core structure component A; blending the above-mentioned mass fractions of modified chitosan, modified hydroxylated silica and alum to obtain the core structure component B;

[0093] S4. Blending the core structure component A and the core structure component B in the above-mentioned mass fractions, and wrapping them with the edible substance to obtain the probiotic preparation for promoting the growth of livestock and poultry.

[0094] Example 3

[0095] A probiotic preparation for promoting the growth of livestock and poultry, the probiotic preparation for promoting the growth of livestock and poultry having a core-shell structure, comprising the following components in parts by weight:

[0096] Nuclear structure component A:

[0097] Probiotics liquid 40 servings

[0098] 70 copies of culture medium;

[0099] Nuclear structural component B:

[0100] Modified chitosan 15 parts

[0101] 15 parts alum

[0102] 10 parts of modified hydroxylated silica;

[0103] Shell structure:

[0104] Edible substances; 1000 portions;

[0105] The preparation method of the probiotic preparation for promoting livestock and poultry growth comprises the following steps:

[0106] S1. Preparation of modified chitosan: dissolve chitosan in an acetic acid solution with a pH of 5, stir, add sodium hydroxide solution to a pH of 10 to precipitate it, filter with suction, then add glutaraldehyde and aluminum sulfate, pre-react in a water bath at 85°C for 1.5h, then add aminodecapolyethylene glycol and continue to react for 4h, precipitate with acetone after the reaction is completed, filter with suction, then wash the product with ethanol, filter with suction, and dry in an oven at 90°C to obtain modified chitosan; wherein the mass ratio of the chitosan, glutaraldehyde, aminodecapolyethylene glycol and aluminum sulfate is 1:5:2:3;

[0107] S2. Preparation of modified hydroxylated silica: 4 parts of hydroxylated silica were added to 15 parts of linoleic acid, ultrasonicated for 30 minutes and heated to 80° C. After the reaction was completed, the mixture was placed in an oven at 50° C. and dried to constant weight to obtain modified hydroxylated silica;

[0108] S3, blending the above-mentioned probiotic solution and culture medium in the above-mentioned mass fractions to obtain the core structure component A; blending the above-mentioned mass fractions of modified chitosan, modified hydroxylated silica and alum to obtain the core structure component B;

[0109] S4. Blending the core structure component A and the core structure component B in the above-mentioned mass fractions, and wrapping them with the edible substance to obtain the probiotic preparation for promoting the growth of livestock and poultry.

[0110] Comparative Example 1

[0111] The raw materials, raw material portions and preparation methods used in Comparative Example 1 are the same as those in Example 1, with the only difference being that in Comparative Example 1, an equal mass portion of modified chitosan is replaced by unmodified chitosan.

[0112] Comparative Example 2

[0113] The raw materials, raw material portions and preparation methods used in Comparative Example 2 are the same as those in Example 1, with the only difference being that in Comparative Example 2, no modified hydroxylated silica is added.

[0114] Test Case

[0115] The probiotic preparations prepared in Example 1 and Comparative Examples 1-2 were subjected to performance tests to compare the differences in performance of the probiotic preparations.

[0116] Test method: In three adjacent duck ponds with a water surface area of ​​about 30 mu, the water depth was 1.5 meters, and the water was divided into experimental group and control group 1 and control group 2. 50 ducks of average size and weight were put into each cage, and 200 ml of the probiotic preparation prepared in Example 1 and comparative example 1-2 were used per mu of water surface in the experimental group, control group 1 and control group 2, respectively; the water was not changed during the whole breeding process, and the water loss by evaporation was supplemented with clean fresh water. After 15 days, the pH value, ammonia nitrogen concentration, nitrite concentration, duck survival rate, duck weight gain rate, COD, and transparency were measured.

[0117] The pH value of the water body is measured according to "HJ 1147-2020 Electrode Method for Determining pH Value of Water Quality";

[0118] The ammonia nitrogen content of water was determined according to "HJ 535-2009 Determination of ammonia nitrogen in water quality - Nessler's reagent spectrophotometry";

[0119] The nitrite content of water was determined according to GB 7493-87 Water quality - Determination of nitrite nitrogen - Molecular absorption spectrophotometry;

[0120] The COD of the water body was measured according to "HJ 828-2017 Determination of Chemical Oxygen Demand of Water Quality - Dichromate Method";

[0121] The transparency of the water body is measured according to "HJ 1075-2019 Turbidimeter Method for Determination of Turbidity of Water Quality";

[0122] The test results are shown in Table 1:

[0123] Table 1

[0124]

[0125] It can be seen from the above table that the ammonia nitrogen content in the water body using the probiotic preparation prepared in Example 1 is low, while the ammonia nitrogen content in the water body in Comparative Examples 1 and 2 is high. This is because the probiotic preparation has the advantages of high flocculation rate, small dosage, but high probiotic content. The high content of probiotics is beneficial to the regulation of water quality, thereby regulating the ammonia nitrogen content and nitrite content in the water body, making the water quality stable, which is beneficial to the growth of poultry and livestock.

[0126] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0127] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A probiotic preparation for promoting the growth of livestock and poultry, characterized in that: The probiotic preparation for promoting livestock and poultry growth has a core-shell structure and includes the following components in parts by weight: Nuclear structure component A: 30-40 servings of probiotic liquid 50-70 copies of culture medium; Nuclear structural component B: Modified chitosan 10-15 parts Alum 10-15 parts 5-10 parts of modified hydroxylated silicon dioxide; Shell structure: edible substances; The modified chitosan is a product obtained by cross-linking chitosan with glutaraldehyde and single-terminated amino polyethylene glycol; The modified hydroxylated silicon dioxide is a product of partially grafting fatty acids onto hydroxylated silicon dioxide; The edible substance is a viscous mixture of starch, corn flour, sweet potato flour and water; The preparation method of the probiotic preparation for promoting livestock and poultry growth comprises the following steps: S1. Preparation of modified chitosan: dissolving chitosan in acetic acid solution, stirring and adding alkali to precipitate it; then adding glutaraldehyde and single-terminated amino polyethylene glycol, and aluminum sulfate, and heating to react; S2. Preparation of modified hydroxylated silicon dioxide: adding hydroxylated silicon dioxide into liquid fatty acid, ultrasonicating and heating; S3, blending the probiotic solution and culture medium to obtain the core structure component A; blending the modified chitosan, modified hydroxylated silica and alum to obtain the core structure component B; S4. Blending the core structure component A and the core structure component B, and wrapping them with the edible substance to obtain the probiotic preparation for promoting the growth of livestock and poultry.

2. The probiotic preparation for promoting livestock and poultry growth according to claim 1, characterized in that: The probiotic liquid is a mixed bacterial liquid of Bacillus subtilis, Lactobacillus plantarum and Candida.

3. The probiotic preparation for promoting the growth of livestock and poultry according to claim 1, characterized in that: The culture medium is a mixture of inorganic salts, trace elements and water.

4. The probiotic preparation for promoting the growth of livestock and poultry according to claim 2, characterized in that: In the probiotic solution, The content of Bacillus subtilis is 2-5×10 8 cfu / g; The content of plant lactobacillus is 3-6×10 8 cfu / g; The content of Candida is 1-4×10 8 cfu / g.

5. The probiotic preparation for promoting the growth of livestock and poultry according to claim 1, characterized in that: In step S1, the mass ratio of chitosan, glutaraldehyde and single-terminated amino polyethylene glycol is 1:3-5:1-2.

6. The probiotic preparation for promoting the growth of livestock and poultry according to claim 1, characterized in that: In step S2, the heating temperature is 70-80°C, and the ultrasonic time is 10-30 minutes.

7. The probiotic preparation for promoting the growth of livestock and poultry according to claim 1, characterized in that: In step S1, the pH after adding alkali is 8-10.

8. The probiotic preparation for promoting the growth of livestock and poultry according to claim 1, characterized in that: In the single-terminated amino polyethylene glycol, the number of repeating units is 8-12.

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