Rumen-permeated Schizochytrium preparation, preparation method and application of rumen-permeated Schizochytrium preparation

Abstract

The present application discloses a rumen-passed Schizochytrium preparation, a preparation method of the rumen-passed Schizochytrium preparation, and an application thereof. The rumen-passed Schizochytrium preparation comprises a core material and at least one layer of coating, wherein the core material comprises calcium alginate microspheres embedded with Schizochytrium cells, and the coating is coated on the core material so that the oil is wrapped in the calcium alginate microspheres, thereby improving the stability of unsaturated fatty acids and reducing the risk of unsaturated fatty acids leaking in the rumen; the preparation method comprises firstly subjecting the Schizochytrium fermentation product to calcium alginate microsphere embedding and immobilization treatment, then drying the immobilized fermentation product to obtain the core material, and finally coating the core material to obtain the rumen-passed Schizochytrium preparation, effectively avoiding the problems of "pipeline blockage" and "oxidation and deterioration of Schizochytrium powder", thereby improving the yield of Schizochytrium powder in the drying process and simplifying the preparation process, and the finally prepared rumen-passed Schizochytrium preparation has an ideal hardness.

Description

Technical Field

[0001] The present application relates to the technical field of animal feed, and in particular to a rumen-permeated Schizochytrium algae preparation, a preparation method of the rumen-permeated Schizochytrium algae preparation, and an application of the same. Background Art

[0002] Docosahexaenoic acid (DHA) belongs to the ω-3 polyunsaturated fatty acid family and is essential to the human body. It is found in high concentrations in fish oil. Animal glycerophospholipids contain varying amounts of DHA, which can be produced from α-linolenic acid during metabolism in the body, but the production level is low and is primarily supplemented through food. DHA is a key component of human cell membranes and nervous tissue, and has important physiological functions such as promoting brain development, promoting optic nerve development, and preventing and treating Alzheimer's disease. It also promotes brain cell development in infants and young children, prevents cardiovascular disease, improves blood circulation, and ensures the normal functioning of the nervous system.

[0003] The human body cannot synthesize DHA on its own and needs to supplement DHA through food, such as livestock products rich in DHA. Schizochytrium algae powder is rich in DHA, and the DHA content in livestock products can be increased by feeding livestock with Schizochytrium algae powder. The conventional preparation method of Schizochytrium algae powder is: centrifugal spray drying the Schizochytrium algae fermentation product to obtain Schizochytrium algae powder, which has the following disadvantages: 1. Since the fermentation product contains a large amount of oil, the prepared Schizochytrium algae powder has the problem of high surface oil content, which makes the Schizochytrium algae powder prone to oxidation and deterioration when heated, and is prone to clogging of the pipeline during the centrifugal spray drying process, resulting in a low yield of Schizochytrium algae powder; 2. The fermentation product needs to be repeatedly centrifuged and washed with water to reduce the salt and sugar content in the fermentation product, which has the disadvantage of high energy consumption. At present, the prior art also discloses a variety of preparation methods for Schizochytrium algae powder, as described below:

[0004] (1) Chinese patent application CN112544799A provides a method for preparing rumen-permeated Schizochytrium algae powder, which comprises shearing and mixing a low-melting-point oil, an attractant, an antioxidant, and a core material obtained by treating Schizochytrium fermentation broth, and then melt-spraying the mixture to obtain Schizochytrium algae powder. In other words, the rumen-permeated Schizochytrium algae powder is directly prepared from the fermentation broth.

[0005] (2) In the existing technology regarding whole-cell rumen-passed algae powder, Chinese patent application CN114698738A provides a method for preparing rumen-passed Schizochytrium algae powder, which uses mesoporous silica to load Schizochytrium algae powder, and then uses stearic acid and corn fiber as wall materials to coat the core material; Chinese patent application CN114158651A provides a method for preparing rumen-passed nutritional enhancer granules, which first mixes Schizochytrium algae powder with stearic acid and then coats the granules with stearic acid; Chinese patent application CN111449172A provides a method for preparing rumen-protected Schizochytrium algae powder, which first uses hydrophobic beeswax to granulate the Schizochytrium algae powder and then uses a membrane material to coat the granules. The above three patent applications all coat the dried Schizochytrium algae powder.

[0006] (3) In the existing technology regarding whole-cell rumen-pass algae powder using fermentation broth as the starting material, Chinese patent application CN101999522B provides a method for preparing whole-cell microalgae powder that allows mammals to produce high DHA milk, which comprises first concentrating the fermentation broth and then adding an antioxidant, then adding it to an aqueous phase containing starch and an emulsifier, drying it to form algae powder, and finally coating the algae powder with resin; CN111011621A provides a method for preparing Schizochytrium algae powder, which comprises first mixing Schizochytrium algae cells, wall materials, emulsifiers and antioxidants, stirring them, shearing and homogenizing them to obtain a suspension, then drying the suspension to obtain Schizochytrium algae powder, and finally coating the Schizochytrium algae powder. The preparation methods of the above two patent applications both add a large amount of auxiliary materials such as wall materials and emulsifiers, which not only increases the production cost, but also leads to a decrease in the strength and yield of the product.

[0007] Therefore, how to provide a Schizochytrium preparation with low manufacturing cost, ideal hardness and not easy to oxidize and deteriorate is the technical problem to be solved by this application. Summary of the Invention

[0008] The present application provides a rumen-permeated Schizochytrium preparation, a preparation method of the rumen-permeated Schizochytrium preparation, and an application thereof, so as to provide a Schizochytrium preparation with low manufacturing cost, ideal hardness, and not easy to oxidize and deteriorate.

[0009] In a first aspect, the present application provides a rumen-permeable Schizochytrium preparation, comprising:

[0010] a core material comprising calcium alginate microspheres embedded with Schizochytrium cells; and

[0011] At least one layer of film is coated on the core material.

[0012] Optionally, the rumen-permeated Schizochytrium preparation consists of the core material and at least one layer of the envelope, and the core material is calcium alginate microspheres embedded with Schizochytrium cells;

[0013] And / or, the average particle size of the calcium alginate microspheres is 10 μm to 1000 μm.

[0014] Optionally, the coating material comprises one or more of a coating agent, an attractant, an antioxidant and an emulsifier.

[0015] Optionally, the coating agent comprises a fat having a melting point below 80°C;

[0016] and / or, the phagostimulant comprises phospholipid;

[0017] And / or, the antioxidant includes one or more of ethoxyquinoline, butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, tert-butylhydroquinone, tea polyphenols, vitamin E, ascorbyl palmitate, phytic acid, rosemary and isoascorbic acid;

[0018] And / or, the emulsifier includes one or more of glyceryl monostearate, glyceryl distearate, Tween, Span, sorbitan fatty acid ester, sucrose fatty acid ester, sodium alginate, gum arabic and xanthan gum.

[0019] Optionally, the coating is a single layer, and the material of the coating is one or more of stearic acid, hydrogenated palm oil and palmitic acid.

[0020] Optionally, the mass of the core material accounts for 80% to 90% of the total mass of the Schizochytrium rumen-permeable preparation;

[0021] and / or, the mass of the coating accounts for 10% to 20% of the total mass of the Schizochytrium preparation;

[0022] And / or, the thickness of the envelope is 0.1 mm to 0.6 mm.

[0023] In a second aspect, the present application also provides a method for preparing a rumen-permeated Schizochytrium preparation, the preparation method comprising the following steps:

[0024] providing a mixture comprising alginate and a Schizochytrium fermentation product comprising Schizochytrium cells;

[0025] mixing the mixture with an aqueous solution containing calcium ions to cause a cross-linking and curing reaction to obtain a product, wherein the product includes calcium alginate microspheres embedded with Schizochytrium cells;

[0026] Drying the product to obtain a core material; and

[0027] The core material is subjected to a coating treatment to obtain a Schizochytrium algae preparation.

[0028] Optionally, the concentration of Schizochytrium in the Schizochytrium fermentation product is 100 g / L to 300 g / L, calculated as a percentage by mass and volume;

[0029] and / or, in the mixture, the Schizochytrium algae cells are measured by dry weight, and the mass ratio of the alginate to the Schizochytrium algae cells is 1:(10-20);

[0030] And / or, the step of mixing the mixture with an aqueous solution containing calcium ions comprises the steps of: injecting the mixture into the aqueous solution containing calcium ions at 25° C. to 45° C. and mixing.

[0031] Optionally, in the step of injecting the mixture into the aqueous solution containing calcium ions, the injection rate is 25 L / h to 35 L / h;

[0032] And / or, the injection pressure is 0.2 bar to 0.5 bar.

[0033] In a third aspect, the present application provides the use of the rumen-permeated Schizochytrium preparation as described in any one of the first aspects, or the rumen-permeated Schizochytrium preparation prepared by the preparation method as described in any one of the second aspects, in increasing the DHA content in poultry and livestock products, specifically: feeding poultry and livestock with the rumen-permeated Schizochytrium preparation as described in any one of the first aspects, or the rumen-permeated Schizochytrium preparation prepared by the preparation method as described in any one of the second aspects.

[0034] The present application provides a rumen-permeated Schizochytrium algae preparation, a preparation method of the rumen-permeated Schizochytrium algae preparation, and an application thereof, which have the following technical effects:

[0035] The rumen-passing Schizochytrium preparation includes a core material and at least one layer of coating, wherein the core material includes calcium alginate microspheres embedded with Schizochytrium cells, and the coating is coated on the core material, so that the oil is wrapped in the calcium alginate microspheres, thereby improving the stability of unsaturated fatty acids and reducing the risk of unsaturated fatty acids leaking in the rumen. The rumen-passing rate of the Schizochytrium preparation can reach more than 96%, and the calcium alginate microspheres and the coating give the rumen-passing Schizochytrium preparation an ideal hardness; in addition, because the oil is wrapped in the calcium alginate microspheres and will not remain on the surface of the rumen-passing Schizochytrium preparation, the rumen-passing Schizochytrium preparation is not prone to oxidation and deterioration.

[0036] In the preparation method of the rumen-permeated Schizochytrium preparation, the fermentation product of Schizochytrium is first immobilized by embedding calcium alginate microspheres, and then the immobilized fermentation product is dried to obtain a core material, and finally the core material is coated to obtain the rumen-permeated Schizochytrium preparation, which effectively avoids the problems of "pipeline blockage" and "oxidation and deterioration of Schizochytrium powder", thereby improving the yield of Schizochytrium powder in the drying process, the yield can reach 95%, and simplifying the preparation process. The final rumen-permeated Schizochytrium preparation has an ideal hardness. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The following detailed description of the specific embodiments of the present application in conjunction with the accompanying drawings will make the technical solutions and beneficial effects of the present application apparent.

[0038] Figure 1 This is a schematic flow chart of a method for preparing a rumen-passed Schizochytrium preparation provided in this application.

[0039] Figure 2 This is a photo of the calcium alginate microspheres embedded with Schizochytrium cells prepared in step S1.3 of Example 1 of the present application. DETAILED DESCRIPTION

[0040] The technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of this application. The experimental methods in the following embodiments that do not specify specific conditions are generally implemented according to conventional technical means in this field, or implemented according to the conditions recommended by the manufacturer.

[0041] Various embodiments in this application may be presented in the form of a range; it should be understood that the description in a range format is only for convenience and brevity and should not be construed as a hard limitation on the scope of this application; therefore, the range description should be considered to have specifically disclosed all possible subranges and single numbers within the range. For example, the description of a range from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as single numbers within the range, such as 1, 2, 3, 4, 5, and 6, regardless of the range. Whenever a numerical range is indicated herein, it is intended to include any cited numeral (fractional or integer) within the indicated range.

[0042] Unless otherwise defined or indicated, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art, and the materials and reagents used in the Examples, Comparative Examples, and Experimental Examples of this application are commercially available. In addition, any methods and materials similar or equivalent to those described herein can be applied to this application. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0043] In this application, the terms "comprises," "comprising," "having," "containing," or similar expressions are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0044] In this application, the term "at least one" refers to one or more. The terms "at least one" and "any of the following" refer to any combination of these, including any combination of a single or multiple types. For example, "at least one of a, b, or c" or "at least one of a, b, and c" can be expressed as: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can each be a single type or multiple types.

[0045] In the present application, "plurality" refers to two or more, for example, two, three, four, etc.

[0046] In this application, "and / or" means that the selection range includes any one of two or more related listed items, and also includes any and all combinations of the related listed items, and the said any and all combinations include any two related listed items, any more related listed items, or the combination of all related listed items. For example, "A and / or B" includes three parallel solutions of A, B and A+B. For another example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the four-item combination of A, B, C, and D (that is, the technical solution connected by "logical AND").

[0047] In this application, “first”, “second”, “third” or similar expressions are used for descriptive purposes only and should not be understood as indicating or implying relative importance, nor as indicating or implying a sequence of precedence.

[0048] In the present application, "schizochytrium" refers to Schizochytrium, a type of marine fungus belonging to the Labyrinthulomycota, Labyrinthulomycota, Thraustochytriales and Thraustochytridaceae, which is a single-cell structure with a spherical shape.

[0049] The present application studies a rumen-passing Schizochytrium preparation, a preparation method for the rumen-passing Schizochytrium preparation, and an application thereof. The rumen-passing Schizochytrium preparation comprises a core material and at least one layer of coating, wherein the core material comprises calcium alginate microspheres embedded with Schizochytrium cells, and the coating is coated on the core material. In the rumen-passing Schizochytrium preparation of the present application, the Schizochytrium cells are embedded in the calcium alginate microspheres, so that the oil is wrapped in the calcium alginate microspheres, effectively improving the problem of the Schizochytrium powder in the prior art being prone to oxidation and deterioration due to the high surface oil content, and improving the stability of unsaturated fatty acids (such as DHA) and reducing the risk of unsaturated fatty acids leaking in the rumen. The rumen-passing rate of the rumen-passing Schizochytrium preparation can reach more than 96%, and the calcium alginate microspheres and the coating give the rumen-passing Schizochytrium preparation an ideal hardness.

[0050] It should be noted that in the prior art, rumen-pass formulations can be made using unsaturated fatty acids (e.g., DHA), coating agents (e.g., stearic acid), and other excipients. When the mass percentage of unsaturated fatty acids is 5%, the mass percentage of the coating agent can reach 90%, resulting in an ideal hardness for the rumen-pass formulation. However, when the rumen-pass formulation is a Schizochytrium algae-coated product, the coating directly coats the Schizochytrium algae cells, which displace the coating. For example, if the mass percentage of unsaturated fatty acids in the Schizochytrium algae-coated product is 5%, the mass percentage of the coating agent is approximately 70%, resulting in poor hardness for the Schizochytrium algae-coated product. In the embodiments of the present application, the coating coats the calcium alginate microspheres embedded with Schizochytrium algae cells. The calcium alginate microspheres embedded with the Schizochytrium algae cells have an ideal hardness, so even if the mass percentage of the coating does not reach 90%, the rumen-pass Schizochytrium algae formulation still has an ideal hardness.

[0051] In some embodiments of the present application, the Schizochytrium algae preparation is composed of the core material and at least one layer of the coating, and the core material is calcium alginate microspheres embedded with Schizochytrium algae cells.

[0052] In some embodiments of the present application, the average particle size of the calcium alginate microspheres is 10 μm to 1000 μm, for example, 10 μm to 50 μm, 50 μm to 100 μm, 100 μm to 500 μm, or 500 μm to 1000 μm.

[0053] In some embodiments of the present application, the coating material comprises one or more of a coating agent, an attractant, an antioxidant, and an emulsifier.

[0054] In some embodiments of the present application, the coating agent includes a fat with a melting point below 80° C., and the coating agent includes but is not limited to one or more of stearic acid and hydrogenated palm oil.

[0055] In some embodiments of the present application, the phagostimulant comprises a phospholipid.

[0056] In some embodiments of the present application, the antioxidant includes one or more of ethoxyquin, butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, tert-butylhydroquinone, tea polyphenols, vitamin E, ascorbyl palmitate, phytic acid, rosemary and isoascorbic acid.

[0057] In some embodiments of the present application, the emulsifier includes one or more of glyceryl monostearate, glyceryl distearate, Tween, Span, sorbitan fatty acid ester, sucrose fatty acid ester, sodium alginate, gum arabic and xanthan gum.

[0058] As an example, the coating is a single layer, that is, the coating is a coating, and the material of the coating is one or more of stearic acid, hydrogenated palm oil and palmitic acid.

[0059] In some embodiments of the present application, the mass of the core material accounts for 80% to 90% of the total mass of the Schizochytrium spp. preparation, for example, it can be 80% to 81%, 81% to 82%, 82% to 83%, 83% to 84%, 84% to 85%, 85% to 86%, 86% to 87%, 87% to 88%, 88% to 89%, or 89% to 90%.

[0060] In some embodiments of the present application, the mass of the envelope accounts for 10% to 20% of the total mass of the Schizochytrium spp. preparation, for example, it can be 10% to 11%, 11% to 12%, 12% to 13%, 13% to 14%, 14% to 15%, 15% to 16%, 16% to 17%, 17% to 18%, 18% to 19%, or 19% to 20%.

[0061] In some embodiments of the present application, the thickness of the envelope is 0.1 mm to 0.6 mm, for example, 0.1 mm to 0.2 mm, 0.2 mm to 0.3 mm, 0.3 mm to 0.4 mm, 0.4 mm to 0.5 mm, or 0.5 mm to 0.6 mm.

[0062] The present application also provides a method for preparing a rumen-passed Schizochytrium preparation, such as Figure 1 As shown, the following steps are included:

[0063] S1. providing a mixture comprising alginate and a Schizochytrium umbellatum fermentation product, wherein the Schizochytrium umbellatum fermentation product comprises Schizochytrium umbellatum cells;

[0064] S2. mixing the mixture of step S1 with an aqueous solution containing calcium ions to cause a cross-linking and curing reaction to obtain a product, wherein the product includes calcium alginate microspheres embedded with Schizochytrium cells;

[0065] S3, drying the product to obtain a core material;

[0066] S4. The core material is coated to obtain the rumen-permeated Schizochytrium preparation.

[0067] It should be noted that the conventional preparation method of Schizochytrium algae powder is to centrifugally spray-dry the Schizochytrium algae fermentation product to obtain Schizochytrium algae powder. The disadvantages of this preparation method are: first, the fermentation product needs to be subjected to repeated centrifugation and water washing processes to reduce the salt and sugar content in the fermentation product, which has the disadvantages of complicated preparation process and high energy consumption; second, since the fermentation product contains a large amount of oil, the Schizochytrium algae powder obtained after the fermentation product is spray-dried has a high surface oil content, and is prone to pipe clogging during the spray drying process; third, during the spray drying process and the subsequent coating process, the Schizochytrium algae powder is prone to oxidation and deterioration. In the preparation method of the embodiment of the present application, the fermentation product of Schizochytrium spp. is first immobilized by embedding calcium alginate microspheres, and then the immobilized fermentation product is dried to obtain a core material, and finally the core material is coated to obtain a rumen-permeated Schizochytrium spp. preparation, which effectively avoids the problems of "blocking the pipeline" and "oxidation and deterioration of Schizochytrium spp. powder", thereby improving the yield of Schizochytrium spp. powder in the drying process and simplifying the preparation process. The final rumen-permeated Schizochytrium spp. preparation has an ideal hardness.

[0068] Specifically, in step S1, the alginate includes, but is not limited to, one or more of sodium alginate and potassium alginate, with sodium alginate being an example. To facilitate the dissolution of the alginate, the alginate can be dissolved in water to produce an alginate aqueous solution, which is then mixed with the fermentation product of Schizochytrium schizochytrium to obtain a mixture. The alginate aqueous solution can be prepared by dissolving an appropriate amount of alginate in water to obtain an alginate aqueous solution. In some embodiments of the present application, the concentration of alginate in the alginate aqueous solution is 10 g / L to 100 g / L.

[0069] In order to improve the embedding effect of Schizochytrium cells, in some embodiments of the present application, step S1 includes the step of adding an alginate aqueous solution to the Schizochytrium fermentation product under stirring.

[0070] In addition, when the volume of the fermentation broth to be processed is large, in order to avoid the addition of alginate further expanding the volume of the fermentation broth and thus prolonging the spraying process time, the alginate can be directly dissolved in the fermentation broth under stirring conditions. In other embodiments, step S1 includes the step of slowly dispersing the alginate into the Schizochytrium fermentation product under stirring conditions.

[0071] In step S2, the "Schizochytrium fermentation product" refers to the culture obtained by inoculating Schizochytrium into a liquid culture medium and fermenting it for a specific period of time. The culture may be directly mixed with the alginate aqueous solution without undergoing any other separation and purification steps, or the culture may be simply concentrated and then mixed with the alginate aqueous solution. In the embodiments of the present application, the "Schizochytrium fermentation product" may be a shake flask fermentation product or a fermentor fermentation product.

[0072] To improve the encapsulation effect of Schizochytrium cells, in some embodiments of the present application, the Schizochytrium cell concentration in the Schizochytrium fermentation product is 100 g / L to 300 g / L, calculated as mass volume percentage, for example, 100 g / L to 150 g / L, 150 g / L to 200 g / L, 200 g / L to 250 g / L, or 250 g / L to 300 g / L. It should be noted that, in the present application, "cell concentration" is measured on a dry weight basis.

[0073] In order to ensure that the Schizochytrium cells in the Schizochytrium fermentation product are fully embedded in calcium alginate, in some embodiments of the present application, in the mixture, the Schizochytrium cells are measured by dry weight, and the mass ratio of alginate to Schizochytrium cells is 1:(10-20), for example, it can be 1:(10-12), 1:(12-14), 1:(14-16), 1:(16-18), or 1:(18-20).

[0074] In step S2, the aqueous solution containing calcium ions may be prepared by dissolving an appropriate amount of a calcium salt in water to obtain an aqueous solution containing calcium ions. The calcium salt includes, but is not limited to, calcium halide and calcium nitrate, and an exemplary calcium salt is calcium chloride. In some embodiments of the present application, the concentration of calcium ions in the aqueous solution containing calcium ions is 1 g / L to 10 g / L, for example, 1 g / L to 3 g / L, 3 g / L to 5 g / L, 5 g / L to 7 g / L, or 7 g / L to 10 g / L.

[0075] To further enhance the encapsulation efficiency of Schizochytrium cells, in some embodiments of the present application, step S2 includes the step of injecting the mixture into an aqueous solution containing calcium ions at a temperature between 25°C and 45°C and mixing. Injection methods include, but are not limited to, injection with a sharp-orifice spray gun and dropwise addition. It should be noted that the crosslinking and curing reaction temperature can be selected based on actual needs, for example, room temperature. The crosslinking and curing reaction time is determined by the injection rate of the mixture. After the mixture is injected, the entire reaction system can be left to stand for a predetermined period of time.

[0076] In order to obtain a more ideal embedding morphology, in some embodiments of the present application, the injection rate is 25L / h to 35L / h, for example, it can be 25L / h to 28L / h, 28L / h to 30L / h, 30L / h to 32L / h, or 33L / h to 35L / h; and / or, the injection pressure is 0.2bar to 0.5bar, for example, it can be 0.2bar to 0.3bar, 0.3bar to 0.4bar, or 0.4bar to 0.5bar.

[0077] In step S3, "drying treatment" includes all processes that can enable the product to obtain higher energy and be converted into a solid. An example of drying treatment is centrifugal spray drying. An example of process parameters for centrifugal spray drying is: inlet air temperature of 180°C to 200°C, outlet air temperature of 85°C to 90°C, and feed rate of 260kg / h to 350kg / h. It should be noted that before the drying treatment, the average particle size of the calcium alginate microspheres can reach 2000μm to 5000μm. Compared with the calcium alginate microspheres before the drying treatment, the average particle size of the calcium alginate microspheres after the drying treatment is reduced, that is, a "shrinkage" phenomenon occurs. The average particle size of the calcium alginate microspheres after the drying treatment can be controlled at 10μm to 1000μm.

[0078] In step S4, "coating" refers to forming one or more functional layers on the surface of the core material. These functional layers have important functions, such as shaping, protecting the active ingredient, improving stability, solubilizing, assisting dissolution, and providing controlled release of the active ingredient. This ensures that the Schizochytrium rumen-permeable preparation has a certain hardness, shelf life, and bioavailability, thereby improving the safety and efficacy of the Schizochytrium rumen-permeable preparation. Materials for the functional layers include, but are not limited to, one or more coating agents, attractants, antioxidants, and emulsifiers. Coating methods include, but are not limited to, at least one of bottom spray coating, fluidized bed coating, and roller coating.

[0079] The embodiments of the present application also provide the use of any of the above-described Schizochytrium preparations, or the Schizochytrium preparations prepared by any of the above-described preparation methods, in increasing the DHA content in poultry and livestock products, specifically: feeding poultry and livestock with any of the above-described Schizochytrium preparations, or the Schizochytrium preparations prepared by any of the above-described preparation methods.

[0080] Among them, "poultry and livestock products" include but are not limited to dairy products, meat products and egg products. "Poultry and livestock" include but are not limited to sheep, cattle, horses, pigs, rabbits, dogs, chickens, ducks and geese.

[0081] I. Description of the Schizochytrium fermentation products involved in the examples of this application:

[0082] (1) Origin of Schizochytrium

[0083] Schizochytrium is a strain owned by Jiabiyu. The name of the preservation institution is: China Center for Type Culture Collection, the preservation institution address is: Wuhan University, Wuhan, China, the preservation number is: CCTCC NO: M 2019990, the classification name is: Schizochytrium, and the preservation date is: December 2, 2019.

[0084] (2) Preparation of Schizochytrium fermentation products

[0085] The Schizochytrium fermentation product was prepared with reference to Example 4 in Chinese patent application CN111235035A. The Schizochytrium fermentation product was a fermentation broth obtained by fermentation culture (without post-treatment).

[0086] II. Method for detecting the rumen release rate of the test samples involved in the experimental examples of this application

[0087] The in vitro artificial simulated digestive fluid method was used to detect the rumen release rate of the test sample (the rumen-permeable Schizochytrium preparations prepared in the examples and comparative examples), comprising the following steps:

[0088] S100. Accurately weigh 10 g of the test sample and place it at the bottom of a 50 mL stoppered test tube.

[0089] S200. Add 20 mL of artificial simulated rumen digestion fluid to the stoppered test tube containing the test sample, cover the test tube stopper tightly, and digest in a constant temperature water bath shaker at 39°C for 0 h, 12 h, 24 h and 48 h. Sampling is performed at 12 h, 24 h and 48 h, respectively. After rinsing and filtering, the filtrate is collected and the volume of the filtrate is adjusted to 5 mL. The content of the active ingredient DHA in the filtrate is determined.

[0090] The pH of the artificial simulated rumen digestive fluid is 6.6. The preparation method of the artificial simulated rumen digestive fluid is as follows: weigh 5.72 g of citric acid and 52.09 g of disodium hydrogen phosphate, dissolve the two in a small amount of distilled water, and adjust the volume to 1000 mL.

[0091] Rumen passage rate of the test sample (W1) = A n / A0×100%, where A0 is the content of DHA in the test sample (undigested), A n It is the DHA content of the test sample when it is digested for a specific time.

[0092] III. Method for detecting the yield of Schizochytrium algae powder involved in the experimental examples of this application

[0093] Since the largest loss in yield occurs mainly in the drying process and the loss in the coating process is very small, the yield of the test samples tested in the experimental examples of this application refers to the yield after completing the drying process. The reason why the drying process causes yield loss is that during the drying process, oil will seep out and cause the wall to stick, and it is necessary to manually scrape off the Schizochytrium powder that sticks to the wall. In subsequent evaluations, the manually scraped Schizochytrium powder often shows characteristics such as easy oxidation and a strong fishy smell.

[0094] The calculation formula of the yield of the test sample is: yield of the test sample = weight of Schizochytrium algae powder actually obtained without manual scraping / (volume of fermentation liquid×cell concentration).

[0095] IV. Hardness Evaluation Method of Test Samples in Experimental Examples of This Application

[0096] Take 10 particles of the sample to be evaluated (basically the same size) and place them in a watch glass. Then move the watch glass into a desiccator filled with a saturated magnesium nitrate solution and keep it at a constant temperature of 25±1℃ for 24 hours. Then take it out and immediately place it one by one into a glass tube (with an inner diameter of 24mm and a length of 200mm) standing on a wooden board (thickness 2cm). Take a cylindrical weight (made of polytetrafluoroethylene, with a diameter of 22mm and a mass of 20±0.1g) and let it fall freely from the mouth of the glass tube. Observe whether the particles are broken and record the number of broken particles. The number of broken particles is used to indicate the friability.

[0097] V. Odor Evaluation Method of Test Samples in Experimental Examples of This Application

[0098] Prior research has revealed that cattle are selective about the fishy odor of feed, and feed with a strong fishy smell can affect their consumption. The applicant has a standard process for evaluating odor sensory perception, using standard samples. Ten evaluators score the fishy odor intensity of the samples on a 7-point scale (absent = 0, somewhat present = 1, mild = 2, moderate = 3-4, relatively strong = 5-6, and very strong = 7). The average of the 10 evaluators' scores represents the fishy odor intensity.

[0099] The technical solutions and beneficial effects of the present application are further illustrated below with reference to embodiments, comparative examples and experimental examples.

[0100] Example 1

[0101] This embodiment provides a rumen-permeated Schizochytrium preparation and a preparation method thereof. The rumen-permeated Schizochytrium preparation is a granule, and the rumen-permeated Schizochytrium preparation consists of a core material and a layer of coating. The core material is calcium alginate microspheres embedded with Schizochytrium cells, and the coating material is palmitic acid.

[0102] The preparation method of the rumen-permeated Schizochytrium preparation comprises the following steps:

[0103] S1.1. Dissolve 10 kg of sodium alginate in tap water by shearing to obtain a 50 g / L sodium alginate aqueous solution.

[0104] S1.2. Providing 10 L of a Schizochytrium fermentation product, wherein the Schizochytrium fermentation product has a Schizochytrium cell concentration of 200 g / L calculated as a mass volume percentage, adding 2000 mL of a sodium alginate aqueous solution to the Schizochytrium fermentation product under stirring, and mixing to obtain a mixture;

[0105] S1.3. Provide a calcium chloride aqueous solution (calcium ion concentration is 1 g / L). At 39°C, inject the mixture prepared in step S1.2 into 2000 mL of calcium chloride aqueous solution through a sharp-hole spray gun at an injection rate of 30 L / h (0.2 bar pressure). Then, let it stand at room temperature (20°C) for 30 minutes to undergo a cross-linking and curing reaction to obtain a product comprising calcium alginate microspheres (such as Figure 2 shown);

[0106] S1.4. Centrifugal spray drying the product obtained in step S1.3 to obtain a core material. The process parameters for centrifugal spray drying are as follows: inlet air temperature of 200°C, outlet air temperature of 90°C, feed rate of 300 kg / h, blower power of 35 Hz (50 Hz at full load), induced draft fan power of 47 Hz (50 Hz at full load), and tower pressure of -100 Pa.

[0107] S1.5. The core material of step S1.4 is coated by bottom spray coating. The coating agent is palmitic acid. The process parameters of the bottom spray coating process are: inlet air temperature is 45°C, material temperature is 32°C, induced draft fan power is 15HZ, feed speed is 2r / min, and spray pressure is 0.2MPa to obtain the rumen-permeable Schizochytrium preparation.

[0108] Example 2

[0109] This embodiment provides a rumen-permeated Schizochytrium preparation and a preparation method thereof. Compared with the preparation method of the rumen-permeated Schizochytrium preparation in Example 1, the preparation method of the rumen-permeated Schizochytrium preparation in this embodiment is different in that: in step S1.2, "adding 2000 mL of sodium alginate aqueous solution to the Schizochytrium fermentation product" is replaced by "adding 4000 mL of sodium alginate aqueous solution to the Schizochytrium fermentation product", and step S1.3 is replaced by "providing a calcium chloride aqueous solution (calcium ion concentration of 5 g / L), injecting the mixture prepared in step S1.2 into 2000 mL of calcium chloride aqueous solution through a sharp-hole spray gun at 39°C, at an injection rate of 25 L / h (0.2 bar pressure), and then standing at room temperature (20°C) for 30 minutes to perform a cross-linking and curing reaction to obtain a product, wherein the product comprises calcium alginate microspheres embedding Schizochytrium cells."

[0110] Example 3

[0111] This embodiment provides a rumen-permeated Schizochytrium preparation and a preparation method thereof. Compared with the preparation method of the rumen-permeated Schizochytrium preparation in Example 1, the preparation method of the rumen-permeated Schizochytrium preparation in this embodiment is different in that: in step S1.2, "adding 2000 mL of sodium alginate aqueous solution to the Schizochytrium fermentation product" is replaced by "adding 1000 mL of sodium alginate aqueous solution to the Schizochytrium fermentation product."

[0112] Example 4

[0113] This embodiment provides a rumen-permeated Schizochytrium preparation and a preparation method thereof. Compared with the preparation method of the rumen-permeated Schizochytrium preparation in Example 1, the preparation method of the rumen-permeated Schizochytrium preparation in this embodiment is different in that: step S1.3 is replaced by "providing a calcium chloride aqueous solution (calcium ion concentration is 0.5 g / L), and at 39°C, injecting the mixture prepared in step S1.2 into 2000 mL of the calcium chloride aqueous solution through a sharp-hole spray gun at an injection rate of 30 L / h (0.2 bar pressure), and then standing at room temperature (20°C) for 30 minutes to perform a cross-linking and curing reaction to obtain a product, wherein the product comprises calcium alginate microspheres embedding Schizochytrium cells."

[0114] Example 5

[0115] This embodiment provides a Schizochytrium algae preparation and a preparation method thereof. Compared with the preparation method of the Schizochytrium algae preparation in Example 1, the preparation method of the Schizochytrium algae preparation in this embodiment is different in that: step S1.3 is replaced by "providing a calcium chloride aqueous solution (calcium ion concentration is 1 g / L), and at 39°C, injecting the mixture prepared in step S1.2 into 2000 mL of the calcium chloride aqueous solution through a sharp-hole spray gun at an injection rate of 50 L / h (1 bar pressure), and then standing at room temperature (20°C) for 30 minutes to perform a cross-linking and curing reaction to obtain a product, wherein the product comprises calcium alginate microspheres embedding Schizochytrium algae cells."

[0116] The product is in strip form and needs to be crushed after drying before the next coating step.

[0117] Example 6

[0118] This embodiment provides a Schizochytrium algae preparation and a preparation method thereof. Compared with the preparation method of the Schizochytrium algae preparation in Example 1, the preparation method of the Schizochytrium algae preparation in this embodiment is different in that: step S1.3 is replaced by "providing a calcium chloride aqueous solution (calcium ion concentration is 1 g / L), and at 39°C, injecting the mixture prepared in step S1.2 into 2000 mL of the calcium chloride aqueous solution through a sharp-hole spray gun at an injection rate of 10 L / h (0.1 bar pressure), and then standing at room temperature (20°C) for 30 minutes to perform a cross-linking and curing reaction to obtain a product, wherein the product comprises calcium alginate microspheres embedding Schizochytrium algae cells."

[0119] Among them, the product has a "wire-drawing" phenomenon and is difficult to shape. After drying, it needs to be crushed before the next coating step can be carried out.

[0120] Comparative Example 1

[0121] This comparative example provides a Schizochytrium algae preparation and a preparation method thereof. The Schizochytrium algae preparation is a granule. The preparation method of the Schizochytrium algae preparation comprises the following steps:

[0122] S10.1. Provide 10,000 mL of a Schizochytrium salina fermentation product, wherein the Schizochytrium salina cell concentration in the Schizochytrium salina fermentation product is 200 g / L, calculated as mass volume percentage. Centrifugal spray drying the Schizochytrium salina fermentation product to obtain a core material. The centrifugal spray drying process parameters are as follows: an inlet air temperature of 200° C., an outlet air temperature of 90° C., a feed rate of 300 kg / h, a forced draft fan power of 35 Hz (50 Hz at full load), an induced draft fan power of 47 Hz (50 Hz at full load), and a tower pressure of -100 Pa.

[0123] S10.2. The core material of step S10.1 is coated by bottom spray coating. The coating agent is palmitic acid. The process parameters of the bottom spray coating process are: inlet air temperature is 45°C, material temperature is 32°C, induced draft fan power is 15HZ, feed speed is 2r / min, and spray pressure is 0.2MPa to obtain the rumen-permeable Schizochytrium preparation.

[0124] Comparative Example 2

[0125] This comparative example provides a Schizochytrium algae preparation and a preparation method thereof. The Schizochytrium algae preparation is a granule. The preparation method of the Schizochytrium algae preparation comprises the following steps:

[0126] S11.1. Provide 10,000 mL of a Schizochytrium fermentation product, wherein the Schizochytrium fermentation product has a Schizochytrium cell concentration of 200 g / L, calculated as a mass volume percentage. Heat the Schizochytrium fermentation product to 50° C., add 2,000 mL of a gelatin aqueous solution (gelatin concentration of 100 g / L) to the Schizochytrium fermentation product, and disperse uniformly to obtain a mixed system.

[0127] S11.2. Add 500 mL of an aqueous sodium polyphosphate solution (sodium polyphosphate concentration: 50 g / L) to the mixture at 50°C. After mixing, adjust the pH to 4.5 with a 10% by mass aqueous acetic acid solution. Cool the mixture to 4°C and continue stirring for 6 hours to obtain a suspension. During the pH adjustment process and the cooling process thereafter, a coacervate formed by gelatin and sodium polyphosphate coats the surface of the oil droplets to form primary microcapsules.

[0128] S11.3. Washing the suspension with water, and spray drying the washed suspension to obtain a core material. The spray drying process parameters are: an inlet air temperature of 170°C and an outlet air temperature of 80°C.

[0129] S11.4. Refer to step S1.5.

[0130] Comparative Example 3

[0131] This comparative example provides a Schizochytrium algae preparation and a preparation method thereof. The Schizochytrium algae preparation is a granule. The preparation method of the Schizochytrium algae preparation comprises the following steps:

[0132] S12.1. Provide 10,000 mL of a Schizochytrium fermentation product, wherein the Schizochytrium cell concentration in the Schizochytrium fermentation product is 200 g / L, calculated as mass volume percentage. Heat the Schizochytrium fermentation product to 50° C., add 0.05 kg of sodium starch octenylsuccinate, 0.05 kg of whey protein, 0.05 kg of monoglyceride, 0.0005 kg of butylated hydroxyanisole, and 0.0002 kg of vitamin E to the Schizochytrium fermentation product, and stir and shear at 40° C. for 40 min to obtain a suspension.

[0133] S12.2. Spray drying the suspension to obtain a core material. The spray drying process parameters are: an air inlet temperature of 170°C and an air outlet temperature of 80°C.

[0134] S12.3. Refer to step S1.5.

[0135] Experimental Example 1

[0136] The yields of Schizochytrium algae powder in Examples 1 to 6 and Comparative Examples 1 to 3 were tested using the method in (III). The results are shown in Table 1 below:

[0137] Table 1 Summary of the yield of Schizochytrium powder in Examples 1 to 6 and Comparative Examples 1 to 3

[0138]

[0139] As can be seen from Table 1, the yield of Schizochytrium algae powder in Examples 1 and 2 has obvious advantages. The reason is that the Schizochytrium algae cells are embedded in the calcium alginate microspheres, so that the oil is wrapped in the calcium alginate microspheres and does not remain on the surface, thereby reducing the risk of "clogging the pipe" and "sticking to the wall" during the drying process, and effectively improving the yield of Schizochytrium algae powder.

[0140] The yield of Schizochytrium algae powder in Comparative Example 1 was the worst. The reason was that the fermentation product of Schizochytrium algae contained a large amount of oil, which resulted in a high oil content on the surface of the Schizochytrium algae powder after spray drying, making it prone to "pipe clogging" and "wall sticking" problems, resulting in a decrease in the yield of the Schizochytrium algae powder.

[0141] The yield of the Schizochytrium algae powder in Comparative Example 2 was poor because: in Comparative Example 2, the Schizochytrium algae powder was embedded in the form of gelatin complex coacervation, but the strength of gelatin was poor and it was easy to break during the drying process, resulting in oil overflow, thereby reducing the yield of the Schizochytrium algae powder.

[0142] The yield of Schizochytrium powder in Comparative Example 3 was average. The reason was that although a variety of auxiliary materials were used to wrap the Schizochytrium cells in Comparative Example 3 so that the oil was wrapped inside, due to the characteristics of the emulsifier used, the amount did not completely wrap the cells, thereby failing to completely reduce the risks of "clogging the pipe" and "sticking to the wall" during the drying process. In addition, the use of multiple auxiliary materials increased the production cost.

[0143] Experimental Example 2

[0144] The performance of the Schizochytrium rumen-passing preparations in Examples 1 to 6 and Comparative Examples 1 to 3 was tested. The performance test indicators included: hardness, average particle size of individual particles, mass percentage of core material, mass percentage of coating, coating thickness, rumen-passing rate, and odor. The performance test results are shown in Table 2 below:

[0145] Table 2 Summary of performance test results of Schizochytrium rumen-permeable preparations in Examples 1 to 6 and Comparative Examples 1 to 3

[0146]

[0147] As can be seen from Table 1, the comprehensive performance of the rumen-passed Schizochytrium preparations in Examples 1 and 2 is the best, with ideal hardness, suitable particle size, low fishy smell and high rumen-passing rate. Among them, under the premise that the average particle size difference of individual particles is small, the hardness of the rumen-passed Schizochytrium preparations in Examples 1 and 2 is significantly better than that of other examples and comparative examples, effectively improving the problem of "breaking into pieces as soon as crushed" in the existing rumen-passed Schizochytrium preparations, making it easier to store and having an ideal rumen-passing rate; in addition, the rumen-passed Schizochytrium preparations in Examples 1 and 2 have a small fishy smell, which is beneficial for cattle to eat.

[0148] The hardness of the rumen-passed Schizochytrium preparation in Comparative Example 1 was low and the fishy smell was high. The reason was that the coating directly covered the Schizochytrium cells, which squeezed out the content of the coating, resulting in a decrease in hardness. In addition, the fermentation product of Schizochytrium contained a large amount of oil, resulting in a high oil content on the surface of the spray-dried Schizochytrium powder, which enhanced the fishy smell and was not conducive to cattle eating.

[0149] The rumen-passing Schizochytrium preparation in Comparative Example 1 had a low hardness, a high fishy odor intensity, and a poor rumen-passing rate. The reason was that Comparative Example 2 used gelatin complex coacervation to embed the Schizochytrium powder, but the strength of gelatin was poor and it was prone to cracking during the drying process, resulting in oil spillage.

[0150] The comprehensive performance of the Schizochytrium algae preparation in Comparative Example 3 is average, but the preparation of the Schizochytrium algae preparation in Comparative Example 3 requires more auxiliary materials and has a higher production cost.

[0151] The above describes in detail the Schizochytrium rumen-permeable preparation, the preparation method, and the application of the Schizochytrium rumen-permeable preparation provided by this application. This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above examples is only intended to help understand the technical solutions and core concepts of this application. Those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some of the technical features therein with equivalents; these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of this application.

Claims

1. A rumen-passing Schizochytrium preparation, characterized in that: The rumen-passed Schizochytrium preparation comprises: a core material comprising calcium alginate microspheres embedded with Schizochytrium cells; and At least one layer of film, covering the core material; The mass of the core material accounts for 80% to 90% of the total mass of the Schizochytrium rumen-permeable preparation; The preparation method of the rumen-permeated Schizochytrium preparation comprises the following steps: Providing a mixture comprising alginate and a Schizochytrium fermentation product, wherein the Schizochytrium fermentation product comprises Schizochytrium cells; in the mixture, the Schizochytrium cells are measured by dry weight, and the mass ratio of the alginate to the Schizochytrium cells is 1:(10-20); mixing the mixture with an aqueous solution containing calcium ions to generate a cross-linking and curing reaction to obtain a product, wherein the product includes calcium alginate microspheres embedded with the Schizochytrium algae cells; performing centrifugal spray drying on the product to obtain a core material; and The core material is subjected to a coating treatment to obtain a Schizochytrium algae preparation.

2. The rumen-permeable Schizochytrium preparation according to claim 1, characterized in that The rumen-passed Schizochytrium preparation is composed of the core material and at least one layer of the coating, wherein the core material is calcium alginate microspheres embedded with Schizochytrium cells; And / or, the average particle size of the calcium alginate microspheres is 10 μm to 1000 μm.

3. The rumen-permeable Schizochytrium preparation according to claim 1 or 2, characterized in that The coating material comprises one or more of a coating agent, an attractant, an antioxidant and an emulsifier.

4. The rumen-permeable Schizochytrium preparation according to claim 3, characterized in that The coating agent includes oil with a melting point below 80°C; and / or, the phagostimulant comprises phospholipid; And / or, the antioxidant includes one or more of ethoxyquinoline, butylated hydroxyanisole, butylated hydroxytoluene, propyl gallate, tert-butylhydroquinone, tea polyphenols, vitamin E, ascorbyl palmitate, phytic acid, rosemary and isoascorbic acid; And / or, the emulsifier includes one or more of glyceryl monostearate, glyceryl distearate, Tween, Span, sorbitan fatty acid ester, sucrose fatty acid ester, sodium alginate, gum arabic and xanthan gum.

5. The rumen-permeated Schizochytrium preparation according to claim 3, characterized in that The coating is a single layer, and the material of the coating is one or more of stearic acid, hydrogenated palm oil and palmitic acid.

6. The Schizochytrium rumen-passing preparation according to claim 1 or 2, characterized in that The mass of the coating accounts for 10% to 20% of the total mass of the Schizochytrium rumen-permeable preparation; And / or, the thickness of the envelope is 0.1 mm to 0.6 mm.

7. The rumen-permeable Schizochytrium preparation according to claim 1, characterized in that Calculated by mass volume percentage, the concentration of Schizochytrium in the Schizochytrium fermentation product is 100 g / L to 300 g / L; And / or, the step of mixing the mixture with an aqueous solution containing calcium ions comprises the steps of: injecting the mixture into the aqueous solution containing calcium ions at 25° C. to 45° C. and mixing.

8. The rumen-permeable Schizochytrium preparation according to claim 7, characterized in that In the step of injecting the mixture into the aqueous solution containing calcium ions, the injection rate is 25 L / h to 35 L / h; And / or, the injection pressure is 0.2 bar to 0.5 bar.

9. Use of the rumen-permeable Schizochytrium preparation according to any one of claims 1 to 8 for increasing the DHA content in livestock products, characterized in that: Feeding livestock with the rumen-passed Schizochytrium preparation according to any one of claims 1 to 8.

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