Microecological preparation for improving animal immunity and preparation method thereof

Abstract

The invention provides a microecological preparation for improving animal immunity and a preparation method thereof, and belongs to the field of preparation of polysaccharides and derivatives thereof. The preparation method comprises the following steps: preparing the prebiotic extracting solution, preparing the composite carrier, preparing the composite outer wall and preparing the composite preparation. The loading capacity of the prepared composite carrier is 8.9-9.3%, and the loading capacity of the composite carrier is high; the prebiotic synanthrin component in the prepared microecological preparation is slowly released in the artificial intestinal juice; the synanthrin content of the microecological preparation prepared by the method is 94.5% after the microecological preparation is refrigerated at 5 DEG C for 10 months, the synanthrin content of the microecological preparation is 89.7% after the microecological preparation is stored at 25 DEG C for 10 months, the synanthrin content of the microecological preparation is 80.7% after the microecological preparation is stored at 37 DEG C for 10 months, and the storage capacity of the microecological preparation is high.

Description

Technical Field

[0001] This invention provides a microecological preparation for enhancing animal immunity and its preparation method, belonging to the field of polysaccharide and its derivative preparation. Background Technology

[0002] Microecological preparations, as key technological products in the field of green additives, can be divided into three categories based on their composition and function: prebiotics, prebiotics, and mixtures of the two. Prebiotics, as the core functional components in the field of green feed additives, refer to a class of non-digestible substances that are not digested and absorbed by the upper digestive tract of animals, can be directed to the large intestine, and selectively promote the proliferation and metabolism of beneficial bacteria (such as bifidobacteria and lactic acid bacteria) in the intestine. Based on their source and structure, they can be divided into the following categories: First, oligosaccharides (such as fructooligosaccharides, galactooligosaccharides, and isomaltooligosaccharides), which resist the decomposition of digestive enzymes through specific glycosidic bond structures; second, polysaccharides (such as inulin, konjac glucomannan, apple pectin, and spirulina), which achieve targeted delivery to the intestine based on their high molecular chain structure; and third, microbial metabolic derivatives (such as yeast cell wall polysaccharide degradation products and lactic acid bacteria extracellular polysaccharides), which have both nutritional supply and mucosal adhesion assistance functions.

[0003] Compared to probiotics, prebiotics have significant advantages in application: First, there is no risk of live bacteria inactivation, eliminating concerns about activity loss due to high-temperature pelleting and long-term storage of feed, and their stability far exceeds that of live bacteria preparations such as lactic acid bacteria and Bacillus. Second, they have extremely strong compatibility, suitable for different breeds such as pigs, poultry, and ruminants, and have no antagonistic effect on nutrients such as protein, minerals, and vitamins in feed, nor are they affected by trace heavy metal ions or residual antibiotics. Third, they have outstanding green and safe attributes, with core components being natural biological substances that can be metabolized into beneficial products such as short-chain fatty acids (such as acetic acid, propionic acid, and butyric acid) in the animal's body through intestinal microbial metabolism, without the problem of chemical drug additive residues, which aligns with the current policy orientation and market demand for green farming.

[0004] However, due to the structural characteristics of prebiotics themselves and the complexity of the animal gut physiological environment, prebiotics still face many key technical bottlenecks in practical applications: First, prebiotics degrade prematurely in the upper digestive tract. The strong acid environment in the animal's stomach will destroy the glycosidic bond structure of prebiotics, causing a large amount of prebiotics to be converted into monosaccharides and directly absorbed, thus failing to play the role of targeted proliferation of beneficial bacteria. Second, the processing and storage stability is poor. The high temperature during feed pelleting will cause the prebiotic structure to be destroyed, and when the humidity in the storage environment is too high, prebiotics are prone to absorbing moisture and clumping, which not only affects the palatability of the feed, but also further aggravates its degradation.

[0005] To address the aforementioned issues, existing technologies primarily employ physical encapsulation or chemical modification to optimize prebiotics by combining them with external loading materials. This aims to improve the retention rate and stability of prebiotics, as well as enhance their loading capacity. Patent CN112972631A discloses a traditional Chinese medicine composition and its preparation method that improves gastric health and regulates intestinal flora. This patent obtains microcapsules containing prebiotic substances by encapsulating Dendrobium officinale extract, sodium alginate, pectin, and xylooligosaccharides, which can improve gastric health and regulate intestinal flora. However, analysis reveals the following shortcomings in the aforementioned patent's method: First, the patent uses sodium alginate gel as the wall material, which has limited loading capacity, restricting the amount of prebiotics that can be loaded. Second, the microencapsulated formulations prepared using this encapsulation method exhibit low tolerance in humid environments and poor storage resistance. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention provides a microecological preparation for enhancing animal immunity and its preparation method. The prepared microecological preparation can effectively protect the activity of prebiotics, effectively utilize the beneficial effects of polysaccharides, and has a high carrier loading rate.

[0007] To solve the above-mentioned technical problems, the present invention adopts the following technical solution: A method for preparing a microecological preparation to enhance animal immunity includes the following steps: preparing a prebiotic extract, preparing a composite carrier, preparing a composite outer wall, and preparing a composite preparation; The method for preparing the prebiotic extract is as follows: Jerusalem artichoke tubers dried to constant weight are ground with a pulverizer and then sieved. After sieving, Jerusalem artichoke powder is obtained. The Jerusalem artichoke powder is added to deionized water for extraction. The container temperature is controlled at 100-110℃ and the heating time is 2-3 hours. After heating, the filtrate is obtained by filtration. The filtrate is concentrated to 1 / 5 of its original volume. The concentration temperature is 65-70℃ and the vacuum degree is 0.078-0.080 MPa. After concentration, the filtrate is cooled to room temperature to obtain a prebiotic extract containing inulin. The sieve used in the sieving step is a 100-mesh sieve; The inulin content is 35.5 g / 100 mL; The mass ratio of the dried Jerusalem artichoke to deionized water is 9-11:80-85.

[0008] The method for preparing the composite carrier is as follows: wheat bran is added to deionized water, stirred evenly, the pH of the solution is adjusted to 10-11, and heated and stirred in a water bath at 55-65℃ for 1.5-2 hours. After stirring, it is washed until neutral, dried and pulverized to obtain purified wheat bran fiber; natural attapulgite is added to sulfuric acid solution, stirred in a water bath at 75-85℃ for 2-3 hours, filtered, washed until neutral, placed in a calcining furnace, and calcined at 290-300℃ for 2-2.3 hours. After calcination, it is cooled and pulverized to obtain activated attapulgite. Purified wheat bran fiber and activated attapulgite were added to pure water and magnetically stirred at room temperature for 20-30 minutes. After stirring, the mixture was transferred to an ultrasonic reactor and ultrasonically oscillated in an ultrasonic field of 35-45 kHz for 5-6 minutes with an ultrasonic power of 170-180 W. After ultrasonication, tannic acid was added to adjust the pH of the solution to 6.5-7.0, and the reaction was carried out at 28-30℃ for 1-1.5 hours. After the reaction, the mixture was filtered, washed, dried, and sieved to obtain wheat bran fiber-attapulgite composite carrier powder, i.e., composite carrier. The mass ratio of wheat bran to deionized water is 1:15-17; The mass ratio of the natural attapulgite clay to the sulfuric acid solution is 1:20-23; The mass ratio of purified wheat bran fiber, activated attapulgite clay, pure water, and tannic acid is 3-5:1:80-90:0.05-0.1. The activated attapulgite clay has a particle size of 120 mesh; the purified wheat bran fiber has a particle size of 100 mesh. The sulfuric acid solution has a mass concentration of 3-5%; the wheat bran fiber-attapulgite composite carrier powder has a particle size of 100 mesh.

[0009] The method for preparing the composite outer wall is as follows: High-ester pectin powder is added to a container containing deionized water, the container temperature is controlled at 50-60℃, and the mixture is stirred at a speed of 90-110 r / min for 30-60 min. After stirring, calcium lactate solution is added dropwise to the container at a rate of 1 mL / min, with continuous stirring during the addition at a speed of 90-110 r / min for 1.5-2.5 h. After stirring, Ca is obtained. 2+ - High-ester pectin composite gel, i.e., composite outer wall; The mass concentration of the calcium lactate solution is 7-9%; The mass ratio of the high-ester pectin powder, deionized water, and calcium lactate solution is 3-5:75-85:10-15.

[0010] The method for preparing the composite formulation is as follows: wheat bran fiber-attapulgite composite carrier powder, prebiotic extract, and deionized water are added to a container and mixed. The mixture is stirred at 150-200 r / min for 1.5-1.8 h at room temperature. After stirring, the mixture is dried and weighed to obtain the prebiotic-loaded composite carrier. The mixing-drying-weighing steps are repeated according to the above process until the mass difference obtained from three consecutive weighings does not exceed ±0.02 g, finally obtaining a saturated prebiotic-loaded composite carrier. The saturated prebiotic-loaded composite carrier, Ca... 2+ High-ester pectin composite gel, pure water, and HEPES buffer were added to a container. The equipment temperature was controlled at 35-45℃, and the mixture was sonicated in a 35-45kHz ultrasonic field for 5-6 minutes at an ultrasonic power of 170-180W. After sonication, high-ester pectin-Ca was obtained. 2+ -A composite carrier cross-linking formulation that incorporates high-ester pectin-Ca 2+ - The composite carrier cross-linking preparation is freeze-dried under vacuum and then pulverized at low temperature to obtain a composite preparation, namely a microecological preparation; The pH of the HEPES buffer solution is 6.8-8.0; The mass ratio of the wheat bran fiber-attapulgite composite carrier powder, prebiotic extract and deionized water is 18-23:22-28:40-50. The particle size of the composite preparation after low-temperature pulverization is 100 mesh; The composite carrier of saturated prebiotics, Ca 2+ The mass ratio of high-ester pectin complex gel, pure water, and HEPES buffer is 26-32:6-10:55-61:2-5.

[0011] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. After purification, wheat bran fiber is free of starch and protein impurities, exposing a large number of hydroxyl and carboxyl active sites while retaining its network structure. After activation, attapulgite clay forms a porous structure, and the pores can be filled with prebiotics, which improves the ion exchange and adsorption capacity with the outside world. Under the action of stirring and ultrasound, prebiotics enter the network pores and micropores of the carrier and are trapped by the physical structure, which improves the loading capacity of the composite carrier for prebiotics.

[0012] 2. The wheat bran fiber-attapulgite composite carrier prepared by this invention, on the one hand, physically retains and adsorbs prebiotics by purifying the network pores of wheat bran fiber and activating the porous structure of attapulgite; on the other hand, it uses tannic acid to construct a ternary chemical cross-linking network of carrier-tannic acid-prebiotics, anchoring the prebiotics inside the carrier and initially resisting the erosion of high-temperature feed pelleting and the strong acid environment of the animal's stomach; the outer Ca 2+ - The high-ester pectin complex gel further forms a protective barrier. High-ester pectin forms anti-H through intermolecular interactions in gastric acid.+ The corrosive hydrophobic network protects the prebiotics inside from being digested and broken down. After entering the small intestine, pectinase in the small intestine can specifically degrade high-ester pectin, causing the gel structure to disintegrate, and the prebiotics are released into the intestine through the opening of the carrier pores.

[0013] 3. The composite carrier prepared by this invention has a loading capacity of 8.9-9.3%, indicating a high prebiotic loading capacity. The prebiotic content of the microecological preparation prepared by this invention is 94.5% after refrigeration at 5°C for 10 months, 89.7% after storage at 25°C for 10 months, and 80.7% after storage at 37°C for 10 months, indicating strong storage capacity of the microecological preparation. Detailed Implementation

[0014] To provide a clearer understanding of the technical features, objectives, and effects of the present invention, specific embodiments of the present invention are now described.

[0015] Example 1 A method for preparing a microecological preparation to enhance animal immunity includes the following steps: preparing a prebiotic extract, preparing a composite carrier, preparing a composite outer wall, and preparing a composite preparation; (1) Preparation of prebiotic extract Jerusalem artichoke tubers dried to constant weight were ground using a pulverizer and then sieved to obtain Jerusalem artichoke powder. The Jerusalem artichoke powder was added to deionized water for extraction, with the container temperature controlled at 105℃ and the heating time at 2.5h. After heating, the filtrate was obtained by filtration. The filtrate was concentrated to 1 / 5 of its original volume at a temperature of 67℃ and a vacuum degree of 0.079MPa. After concentration, the filtrate was cooled to room temperature to obtain a prebiotic extract containing inulin. The sieve used in the sieving step is a 100-mesh sieve; The inulin content is 35.5 g / 100 mL; The mass ratio of the dried Jerusalem artichoke to deionized water is 10:83.

[0016] (2) Preparation of composite carrier Wheat bran was added to deionized water and stirred evenly. The pH of the solution was adjusted to 10.5, and the mixture was heated and stirred in a 60°C water bath for 1.7 hours. After stirring, the mixture was washed until neutral, dried, and pulverized to obtain purified wheat bran fiber. Natural attapulgite was added to sulfuric acid solution and stirred in a 80°C water bath for 2.5 hours. After filtration and washing until neutral, the mixture was placed in a calcining furnace and calcined at 295°C for 2.1 hours. After calcination, the mixture was cooled and pulverized to obtain activated attapulgite. The purified wheat bran fiber and activated attapulgite were added to pure water and magnetically stirred at room temperature for 25 minutes. After stirring, the mixed solution was transferred to an ultrasonic reactor and ultrasonically oscillated in a 40kHz ultrasonic field for 5.5 minutes at an ultrasonic power of 175W. After ultrasonication, tannic acid was added to adjust the pH of the solution to 6.7, and the mixture was reacted at 29°C for 1.3 hours. After the reaction, the mixture was filtered, washed, dried, and sieved to obtain wheat bran fiber-attapulgite composite carrier powder, i.e., the composite carrier. The mass ratio of wheat bran to deionized water is 1:16; The mass ratio of the natural attapulgite clay to the sulfuric acid solution is 1:22; The mass ratio of purified wheat bran fiber, activated attapulgite clay, pure water, and tannic acid is 4:1:85:0.08. The activated attapulgite clay has a particle size of 120 mesh; the purified wheat bran fiber has a particle size of 100 mesh. The sulfuric acid solution has a mass concentration of 4%; the wheat bran fiber-attapulgite composite carrier powder has a particle size of 100 mesh.

[0017] (3) Preparation of composite outer wall High-ester pectin powder was added to a container containing deionized water, and the container temperature was controlled at 55℃. The mixture was stirred at 100 rpm for 45 min. After stirring, calcium lactate solution was added dropwise to the container at a rate of 1 mL / min, with continuous stirring at 99 rpm for 2.0 h. After stirring, Ca was obtained. 2+ - High-ester pectin composite gel, i.e., composite outer wall; The mass concentration of the calcium lactate solution is 8%; The mass ratio of the high-ester pectin powder, deionized water, and calcium lactate solution is 4:80:13.

[0018] (4) Preparation of compound formulations Wheat bran fiber-attapulgite composite carrier powder, prebiotic extract, and deionized water were added to a container and mixed. The mixture was stirred at 180 r / min for 1.7 h at room temperature. After stirring, the mixture was dried and weighed to obtain the prebiotic-loaded composite carrier. The mixing-drying-weighing steps were repeated until the mass difference between three consecutive weighings did not exceed ±0.02 g, finally obtaining a saturated prebiotic-loaded composite carrier. The saturated prebiotic-loaded composite carrier, Ca... 2+ High-ester pectin composite gel, pure water, and HEPES buffer were added to a container. The equipment temperature was controlled at 40℃, and the mixture was sonicated in a 40kHz ultrasonic field for 5.5 minutes at an ultrasonic power of 175W. After sonication, high-ester pectin-Ca was obtained. 2+ -A composite carrier cross-linking formulation that incorporates high-ester pectin-Ca 2+ - The composite carrier cross-linking preparation is freeze-dried under vacuum and then pulverized at low temperature to obtain a composite preparation, namely a microecological preparation; The pH of the HEPES buffer solution is 6.9; The mass ratio of the wheat bran fiber-attapulgite composite carrier powder, prebiotic extract and deionized water is 20:25:45. The particle size of the composite preparation after low-temperature pulverization is 100 mesh; The composite carrier of saturated prebiotics, Ca 2+ The mass ratio of high-ester pectin complex gel, pure water, and HEPES buffer is 28:8:59:4.

[0019] Example 2 A method for preparing a microecological preparation to enhance animal immunity includes the following steps: preparing a prebiotic extract, preparing a composite carrier, preparing a composite outer wall, and preparing a composite preparation; (1) Preparation of prebiotic extract Jerusalem artichoke tubers dried to constant weight were ground using a pulverizer and then sieved to obtain Jerusalem artichoke powder. The Jerusalem artichoke powder was added to deionized water for extraction, with the container temperature controlled at 100℃ and the heating time at 2 hours. After heating, the filtrate was obtained by filtration. The filtrate was concentrated to 1 / 5 of its original volume at a temperature of 65℃ and a vacuum degree of 0.078 MPa. After concentration, the filtrate was cooled to room temperature to obtain a prebiotic extract containing inulin. The sieve used in the sieving step is a 100-mesh sieve; The inulin content is 35.5 g / 100 mL; The mass ratio of the dried Jerusalem artichoke to deionized water is 9:80.

[0020] (2) Preparation of composite carrier Wheat bran was added to deionized water and stirred evenly. The pH of the solution was adjusted to 10, and the mixture was heated and stirred in a water bath at 55°C for 1.5 hours. After stirring, the mixture was washed until neutral, dried, and pulverized to obtain purified wheat bran fiber. Natural attapulgite was added to sulfuric acid solution and stirred in a water bath at 75°C for 2 hours. After filtration and washing until neutral, the mixture was placed in a calcining furnace and calcined at 290°C for 2 hours. After calcination, the mixture was cooled and pulverized to obtain activated attapulgite. The purified wheat bran fiber and activated attapulgite were added to pure water and magnetically stirred at room temperature for 20 minutes. After stirring, the mixed solution was transferred to an ultrasonic reactor and ultrasonically oscillated in a 35kHz ultrasonic field for 5 minutes at an ultrasonic power of 170W. After ultrasonication, tannic acid was added to adjust the pH of the solution to 6.5, and the mixture was reacted at 28°C for 1 hour. After the reaction, the mixture was filtered, washed, dried, and sieved to obtain wheat bran fiber-attapulgite composite carrier powder, i.e., the composite carrier. The mass ratio of wheat bran to deionized water is 1:15; The mass ratio of the natural attapulgite clay to the sulfuric acid solution is 1:20; The mass ratio of the purified wheat bran fiber, activated attapulgite clay, pure water, and tannic acid is 3:1:80:0.05. The activated attapulgite clay has a particle size of 120 mesh; the purified wheat bran fiber has a particle size of 100 mesh. The sulfuric acid solution has a mass concentration of 3%; the wheat bran fiber-attapulgite composite carrier powder has a particle size of 100 mesh.

[0021] (3) Preparation of composite outer wall High-ester pectin powder was added to a container containing deionized water, and the container temperature was controlled at 50℃. The mixture was stirred at 90 rpm for 30 min. After stirring, calcium lactate solution was added dropwise to the container at a rate of 1 mL / min, with continuous stirring at 90 rpm for 1.5 h. After stirring, Ca was obtained. 2+ - High-ester pectin composite gel, i.e., composite outer wall; The mass concentration of the calcium lactate solution is 7%; The mass ratio of the high-ester pectin powder, deionized water, and calcium lactate solution is 3:75:10.

[0022] (4) Preparation of compound formulations Wheat bran fiber-attapulgite composite carrier powder, prebiotic extract, and deionized water were added to a container and mixed. The mixture was stirred at 150 r / min for 1.5 h at room temperature. After stirring, the mixture was dried and weighed to obtain the prebiotic-loaded composite carrier. The mixing-drying-weighing steps were repeated until the mass difference between three consecutive weighings did not exceed ±0.02 g, finally obtaining a saturated prebiotic-loaded composite carrier. The saturated prebiotic-loaded composite carrier, Ca... 2+ High-ester pectin composite gel, pure water, and HEPES buffer were added to a container. The equipment temperature was controlled at 35℃, and the mixture was sonicated in a 35kHz ultrasonic field for 5 minutes at an ultrasonic power of 170W. After sonication, high-ester pectin-Ca was obtained. 2+ -A composite carrier cross-linking formulation that incorporates high-ester pectin-Ca 2+ - The composite carrier cross-linking preparation is freeze-dried under vacuum and then pulverized at low temperature to obtain a composite preparation, namely a microecological preparation; The pH of the HEPES buffer solution is 6.8; The mass ratio of the wheat bran fiber-attapulgite composite carrier powder, prebiotic extract and deionized water is 18:22:40. The particle size of the composite preparation after low-temperature pulverization is 100 mesh; The composite carrier of saturated prebiotics, Ca 2+ The mass ratio of high-ester pectin complex gel, pure water, and HEPES buffer is 26:6:55:2.

[0023] Example 3 A method for preparing a microecological preparation to enhance animal immunity includes the following steps: preparing a prebiotic extract, preparing a composite carrier, preparing a composite outer wall, and preparing a composite preparation; (1) Preparation of prebiotic extract Jerusalem artichoke tubers dried to constant weight were ground using a pulverizer and then sieved to obtain Jerusalem artichoke powder. The Jerusalem artichoke powder was added to deionized water for extraction, with the container temperature controlled at 110℃ and the heating time at 3 hours. After heating, the filtrate was obtained by filtration. The filtrate was concentrated to 1 / 5 of its original volume at a temperature of 70℃ and a vacuum degree of 0.080 MPa. After concentration, the filtrate was cooled to room temperature to obtain a prebiotic extract containing inulin. The sieve used in the sieving step is a 100-mesh sieve; The inulin content is 35.5 g / 100 mL; The mass ratio of the dried Jerusalem artichoke to deionized water is 11:85.

[0024] (2) Preparation of composite carrier Wheat bran was added to deionized water and stirred evenly. The pH of the solution was adjusted to 11, and the mixture was heated and stirred in a 65°C water bath for 2 hours. After stirring, the mixture was washed until neutral, dried, and pulverized to obtain purified wheat bran fiber. Natural attapulgite was added to sulfuric acid solution and stirred in a 85°C water bath for 3 hours. After filtration and washing until neutral, the mixture was placed in a calcining furnace and calcined at 300°C for 2.3 hours. After calcination, the mixture was cooled and pulverized to obtain activated attapulgite. The purified wheat bran fiber and activated attapulgite were added to pure water and magnetically stirred at room temperature for 30 minutes. After stirring, the mixed solution was transferred to an ultrasonic reactor and ultrasonically oscillated in a 45kHz ultrasonic field for 6 minutes at an ultrasonic power of 180W. After ultrasonication, tannic acid was added to adjust the pH of the solution to 7.0, and the mixture was reacted at 30°C for 1.5 hours. After the reaction, the mixture was filtered, washed, dried, and sieved to obtain wheat bran fiber-attapulgite composite carrier powder, i.e., the composite carrier. The mass ratio of wheat bran to deionized water is 1:17; The mass ratio of the natural attapulgite clay to the sulfuric acid solution is 1:23; The mass ratio of the purified wheat bran fiber, activated attapulgite clay, pure water, and tannic acid is 5:1:90:0.1. The activated attapulgite clay has a particle size of 120 mesh; the purified wheat bran fiber has a particle size of 100 mesh. The sulfuric acid solution has a mass concentration of 5%; the wheat bran fiber-attapulgite composite carrier powder has a particle size of 100 mesh.

[0025] (3) Preparation of composite outer wall High-ester pectin powder was added to a container containing deionized water, and the container temperature was controlled at 60℃. The mixture was stirred at 110 rpm for 60 min. After stirring, calcium lactate solution was added dropwise to the container at a rate of 1 mL / min, with continuous stirring at 110 rpm for 2.5 h. After stirring, Ca was obtained. 2+ - High-ester pectin composite gel, i.e., composite outer wall; The mass concentration of the calcium lactate solution is 9%; The mass ratio of the high-ester pectin powder, deionized water, and calcium lactate solution is 5:85:15.

[0026] (4) Preparation of compound formulations Wheat bran fiber-attapulgite composite carrier powder, prebiotic extract, and deionized water were added to a container and mixed. The mixture was stirred at 200 rpm for 1.8 h at room temperature. After stirring, the mixture was dried and weighed to obtain the prebiotic-loaded composite carrier. The mixing-drying-weighing steps were repeated until the mass difference between three consecutive weighings did not exceed ±0.02 g, finally obtaining a saturated prebiotic-loaded composite carrier. The saturated prebiotic-loaded composite carrier, Ca... 2+ High-ester pectin composite gel, pure water, and HEPES buffer were added to a container. The equipment temperature was controlled at 45℃, and the mixture was sonicated in a 45kHz ultrasonic field for 6 minutes at an ultrasonic power of 180W. After sonication, high-ester pectin-Ca was obtained. 2+ -A composite carrier cross-linking formulation that incorporates high-ester pectin-Ca 2+ - The composite carrier cross-linking preparation is freeze-dried under vacuum and then pulverized at low temperature to obtain a composite preparation, namely a microecological preparation; The pH of the HEPES buffer solution is 8.0; The mass ratio of the wheat bran fiber-attapulgite composite carrier powder, prebiotic extract and deionized water is 23:28:50. The particle size of the composite preparation after low-temperature pulverization is 100 mesh; The composite carrier of saturated prebiotics, Ca 2+ The mass ratio of high-ester pectin complex gel, pure water, and HEPES buffer is 32:10:61:5.

[0027] Comparative Example 1 Based on Example 1, the changes are as follows: no composite outer wall is prepared, no composite preparation is prepared, and wheat bran fiber-attapulgite composite carrier powder, prebiotic extract and deionized water are added to a container and mixed. The mixture is stirred at 180 r / min for 1.7 h at room temperature. After stirring, the mixture is dried and weighed to obtain a prebiotic-loaded composite carrier. The mixing-stirring-drying-weighing steps are repeated according to the above process until the mass difference obtained from three consecutive weighings does not exceed ±0.02 g. Finally, a saturated prebiotic-loaded composite carrier is obtained. The saturated prebiotic-loaded composite carrier is the microecological preparation. The microecological preparation has a particle size of 100 mesh.

[0028] Comparative Example 2 Based on Example 1, the change is that the composite carrier is not prepared. In the step of preparing the composite preparation, the prebiotic extract, the composite outer wall, pure water and HEPES buffer are added to the container, the equipment temperature is controlled at 40°C, and the mixture is ultrasonically oscillated in a 40kHz ultrasonic field for 5.5 minutes with an ultrasonic power of 175W. After the ultrasonication, the prebiotic-embedded gel preparation is obtained. The prebiotic-embedded gel preparation is then vacuum freeze-dried and pulverized at low temperature to obtain the composite preparation, i.e., the microecological preparation. The microecological preparation has a particle size of 100 mesh.

[0029] Test Example 1: Loading capacity test of composite carrier The composite carriers prepared in Examples 1-3 of the same mass were weighed and recorded as the initial mass m1. The composite carrier powder, prebiotic extract and deionized water were added to a container and mixed. The mixture was stirred at 180 r / min for 1.7 h at room temperature. After stirring, the mixture was dried and weighed to obtain the prebiotic-loaded composite carrier. The mixing-stirring-drying-weighing steps were repeated according to the above process until the mass difference of three consecutive weighings did not exceed ±0.02 g. Finally, the saturated prebiotic-loaded composite carrier was obtained, and the mass obtained was recorded as m2. Loading amount = (m2-m1) / m1. The results are shown in Table 1. Table 1. Test results of prebiotic loading on the composite carrier

[0030] Experimental Example 2: Storage Test of Probiotic Preparations Experimental Environment: The microecological preparations obtained in Example 1 and Comparative Examples 1-2 were packaged into sealed bags, each containing 10g (simulating actual packaging). The sealed bags were stored at a constant temperature and protected from light at 5℃, 25℃, 37℃, and 75% relative humidity, respectively. Inulin Content Measurement Method: The microecological preparation samples were hydrolyzed, and the inulin content was tested according to the standard GB / T15672-2009. Inulin content (g / 100g) = [C×V×D / (m×1000)]×100, with the initial inulin content set at 100%. Where C: inulin concentration (mg / mL) obtained from the standard curve; V: volume of the test solution; D: dilution factor (if the concentration of the test solution is too high, it needs to be diluted before measurement; here D=1, undiluted); m: sample mass (g). The test results are shown in Table 2. Table 2.1 Changes in inulin content of the microecological preparation prepared in Example 1 at different temperatures

[0031] Table 2.2 Changes in inulin content of the microecological preparations prepared in Comparative Example 1 at different temperatures

[0032] Table 2.3 Changes in inulin content of the microecological preparations prepared in Comparative Example 2 at different temperatures

[0033] Example 3: Testing of probiotic preparations with artificial gastrointestinal fluid. Simulated artificial gastric fluid: The pH of a 0.2% (w / v) NaCl solution was adjusted to 2.0 with concentrated hydrochloric acid, and pepsin was added to achieve a final concentration of 0.3 g / L. The solution was then filtered through a 0.22 mm aqueous phase filter membrane for sterilization. Simulated artificial intestinal fluid: 1% (w / v) trypsin and 0.3% (w / v) bile salts were dissolved in MRS broth, the pH was adjusted to 6.5, and the mixture was filtered through a 0.22 μm aqueous membrane for sterilization. 3g of the probiotic preparation was placed in conical flasks containing 100mL of simulated gastric fluid and 250mL of simulated intestinal fluid, respectively. The flasks were incubated at 37℃ for 3 hours, and then shaken at 150r / min to dissolve the inulin. The solutions in the conical flasks were then analyzed to determine the inulin content and to assess the solubility in the simulated gastric and intestinal fluids. The results are shown in Table 3. Table 3 Results of Simulated Artificial Gastrointestinal Fluid Test

[0034] As can be seen from Table 1, the loading of the composite carriers prepared in Examples 1-3 is 8.9-9.3%, and the loading of prebiotics in the carriers is high.

[0035] As shown in Table 2.1, the inulin content of the microecological preparation prepared in Example 1 was 94.5% after refrigeration at 5℃ for 10 months, 89.7% after storage at 25℃ for 10 months, and 80.7% after storage at 37℃ for 10 months. Through the dual structure of the composite carrier and the composite outer wall, the prebiotics were adsorbed and fixed by the carrier, while the outer wall isolated moisture and oxygen, exhibiting optimal stability under all storage conditions and the lowest degradation rate within 10 months. Comparative Example 1, lacking the physical barrier of the outer wall, was more susceptible to environmental humidity, resulting in a significantly faster degradation rate than Example 1. Comparative Example 2, lacking the adsorption and fixation of prebiotics by the carrier, allowed inulin to easily diffuse within the gel and come into contact with the outside environment, leading to a faster degradation rate than Comparative Example 1.

[0036] As can be seen from Table 3, the prebiotic inulin in the microecological preparations prepared in Examples 1-3 was not released in the artificial stomach. In Comparative Example 1, without the protection of the outer wall, the prebiotic inulin was released in the stomach. In the microecological preparations prepared in Examples 1-3, the prebiotic inulin was slowly released in the artificial intestinal fluid. In Comparative Example 2, without the loading of the carrier, a large amount of prebiotic was released, and the artificial intestinal fluid contained a large amount of inulin.

[0037] Unless otherwise stated, all percentages used in this invention are mass percentages.

[0038] Finally, it should be noted that the above descriptions are merely preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a microecological preparation for improving immunity of an animal, characterized by, The preparation method comprises the steps of preparing a prebiotic extract, preparing a composite carrier, preparing a composite outer wall, and preparing a composite preparation. The method for preparing the composite carrier comprises the following steps: adding wheat bran into deionized water, adjusting the pH of the solution to 10-11, heating and stirring in a water bath, filtering, drying and crushing after the stirring is completed, to obtain purified wheat bran fiber; adding natural attapulgite into a sulfuric acid solution, stirring in a water bath, filtering, washing to neutral, and calcining, to obtain activated attapulgite; adding the purified wheat bran fiber and the activated attapulgite into pure water, and stirring magnetically, then transferring the mixed solution into an ultrasonic reactor for ultrasonic oscillation, adding tannic acid after the ultrasonic oscillation is completed, adjusting the pH of the solution to 6.5-7.0, filtering, washing, drying and sieving after the reaction is completed, to obtain a wheat bran fiber-attapulgite composite carrier powder, i.e. the composite carrier.

2. The method according to claim 1, wherein the mass ratio of the wheat bran to the deionized water is 1:15-17.

3. The method according to claim 1, wherein the mass ratio of the natural attapulgite to the sulfuric acid solution is 1:20-23.

4. The method according to claim 1, wherein the mass ratio of the purified wheat bran fiber, the activated attapulgite, the pure water and the tannic acid is 3-5:1:80-90:0.05-0.

1.

5. The method according to claim 1, wherein the particle size of the activated attapulgite is 120 mesh, and the particle size of the purified wheat bran fiber is 100 mesh.

6. The method according to claim 1, wherein the mass concentration of the sulfuric acid solution is 3-5%, and the particle size of the wheat bran fiber-attapulgite composite carrier powder is 100 mesh.

3. The method according to claim 1, wherein the method for preparing the prebiotic extract comprises the following steps: grinding the dried Jerusalem artichoke tubers to constant weight with a grinder and sieving, obtaining Jerusalem artichoke powder after the sieving is completed, adding the Jerusalem artichoke powder into deionized water for extraction, controlling the temperature of the container to be 100-110℃, heating for 2-3h, filtering after the heating is completed, concentrating the filtrate, concentrating to 1 / 5 of the original volume of the filtrate, the concentration temperature being 65-70℃, the vacuum degree being 0.078-0.080MPa, cooling to room temperature after the concentration is completed, and obtaining a prebiotic extract containing inulin.

4. The method according to claim 3, wherein the screen used in the sieving step is 100 mesh.

5. The method according to claim 1, wherein the inulin content is 35.5g / 100mL.

6. The method according to claim 1, wherein the mass ratio of the dried Jerusalem artichoke to the deionized water is 9-11:80-85.

5. The method according to claim 1, wherein the method for preparing the composite outer wall comprises the following steps: adding the prebiotic extract into deionized water, adjusting the pH of the solution to 6.5-7.0, adding calcium lactate solution, adjusting the pH of the solution to 7.5-8.5, adding the high-ester pectin powder, adjusting the pH of the solution to 6.5-7.0, and stirring magnetically.

6. The method according to claim 5, wherein the mass concentration of the calcium lactate solution is 7-9%.

7. The method according to claim 5, wherein the mass ratio of the high-ester pectin powder, the deionized water and the calcium lactate solution is 3-5:75-85:10-15. ​ The method for preparing the composite outer wall is as follows: high-ester pectin powder is added into a container containing deionized water, the temperature of the container is controlled at 50-60°C, stirring is carried out at a rotation speed of 90-110 r / min for 30-60 min, after the stirring is completed, calcium lactate solution is added dropwise into the container, the dropping speed is controlled at 1 mL / min, stirring is continuously carried out during the dropping, the stirring rotation speed is 90-110 r / min, the stirring time is 1.5-2.5 h, after the stirring is completed, Ca 2+ - high-ester pectin composite gel, i.e. composite outer wall. ​ ​ ​ 7. The method for preparing the microecological preparation for improving immunity of animals according to claim 1, characterized in that, The method for preparing the composite preparation is as follows: the wheat bran fiber-attapulgite composite carrier powder, prebiotic extract liquid and deionized water are added into a container and mixed, stirred at a speed of 150-200 r / min at room temperature for 1.5-1.8 h, after stirring, dried and weighed to obtain the composite carrier loaded with prebiotics, and the mixing-stirring-drying-weighing steps are repeated according to the above process until the mass difference of three consecutive weighings is not more than ±0.02 g, and finally the composite carrier saturated with prebiotics is obtained; the composite carrier saturated with prebiotics, Ca 2+ - high ester pectin composite gel, pure water, HEPES buffer are added into a container, the equipment temperature is controlled at 35-45℃, ultrasonic oscillation is carried out in a 35-45 kHz ultrasonic field for 5-6 min, the ultrasonic power is 170-180 W, after ultrasonic, high ester pectin-Ca 2+ - composite carrier crosslinked preparation, the high ester pectin-Ca 2+ - composite carrier crosslinked preparation, the high ester pectin-Ca - composite carrier crosslinked preparation, the high ester pectin-Ca 8. The method for preparing the microecological preparation for improving immunity of animals according to claim 7, characterized in that, the pH of the HEPES buffer is 6.8-8.0; the mass ratio of the wheat bran fiber-attapulgite composite carrier powder, the prebiotic extract and the deionized water is 18-23:22-28:40-50; the particle size of the composite preparation after low-temperature pulverization is 100 mesh; The complex carrier of the saturated load prebiotic, Ca 2+ - mass ratio of high-ester pectin complex gel, pure water and HEPES buffer is 26-32:6-10:55-61:2-5.

9. The microecological preparation for improving immunity of animals prepared by the method according to any one of claims 1-8.

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