Feed additive containing compound probiotics and preparation method thereof

Abstract

The invention relates to the field of animal feeding feeds, in particular to a compound probiotic-containing feed additive and a preparation method thereof, and the compound probiotic-containing feed additive comprises the following raw materials: maguey extract, modified pachymaran, compound probiotics, compound vitamins, limonin, acetic acid, beta-amylase, corn and wheat bran. The maguey extract, the modified pachymaran and the compound probiotics play a role through a synergistic effect, the modified pachymaran wraps the compound probiotics and the maguey extract in the feed to form a compound, and the maguey extract provides energy for the compound probiotics, so that the loss process of substances such as the compound probiotics in the feed can be reduced; meanwhile, the effect of the compound probiotics on intestinal epithelial cells is promoted, and nutrient absorption is promoted. The maguey extract, the modified pachymaran and the compound probiotics form a dynamic balance system, and the compound probiotic feed additive which is effective to animal health is provided.

Description

Technical Field

[0001] The invention relates to the field of animal feeding feed, in particular to a feed additive containing composite probiotics and a preparation method thereof. Background Art

[0002] With the rapid development of the breeding industry, the demand for feed additives in livestock and poultry breeding is becoming more and more stringent. Traditional feed additives have problems such as single function, poor stability, and unsatisfactory application effect. These problems not only limit the improvement of animal production performance, but also may pose a potential threat to animal health and food safety. The main hazards of lack of probiotics in feed include impaired intestinal health, imbalance of intestinal flora, weakened epithelial barrier function, abnormal increase in intestinal permeability, thereby increasing the risk of disease, providing colonization opportunities for pathogens, increasing the incidence of diarrhea, and causing animal immunity to decline. At present, the common probiotic feed additives on the market mainly include lactic acid bacteria, bacillus, yeast, etc. Although these single strains have their own characteristics, they cannot comprehensively improve the intestinal health and production performance of animals. Therefore, it is difficult to meet the diverse needs of modern intensive breeding. In addition, in complex environments such as feed processing, storage and animal digestive tract, there are problems such as poor strain adaptability, low survival rate, and poor compatibility of some strains with feed raw materials, which affect product stability. Therefore, the development of a composite probiotic feed additive with stable performance, synergistic functions and good application effect is of great significance to improving breeding benefits and promoting the healthy development of the breeding industry. Summary of the invention

[0003] (1) Technical issues to be resolved The purpose of the present invention is to provide a feed additive containing composite probiotics and a preparation method thereof. The method not only incorporates a variety of composite probiotics into the feed, but also ensures the efficient utilization and survival of these probiotics in the animal intestine by finely screening and optimizing the components of the composite probiotic feed additive, thereby greatly improving the probiotic effect of the probiotics, and finally providing a high-quality composite probiotic feed that is both nutritious and healthy.

[0004] (2) Technical solution To achieve the above object, on the one hand, the present invention provides a feed additive containing composite probiotics, comprising the following raw materials in parts by weight: 10-20 parts of agave extract, 5-15 parts of acetic acid, 1-3 parts of β-amylase, 20-40 parts of composite probiotics, 20-40 parts of composite vitamins, 5-15 parts of limonin, 10-20 parts of taurine, 100-200 parts of corn, and 20-30 parts of wheat bran; The composite probiotic feed additive also includes: Modified pachymaran; The modified pachymaran and the composite probiotics are in a weight ratio of 1:(3-6); The modified pachymaran is prepared by encapsulating pachymaran in a lipid bilayer structure, and the molecular weight of the modified pachymaran is 15000-20000 Da.

[0005] Furthermore, the preparation method of the modified Poria cocos polysaccharide comprises: S11. Add 50-60 g of Poria cocos to 75%-80% ethanol solution under heating and stirring, heat to 60-70°C, stir for 4-5 hours, stir evenly until completely dissolved, and obtain Poria cocos extract; S12. 5 to 6 g of phosphatidylethanolamine was stirred at room temperature, and then a 75% to 80% ethanol solution was added to promote the reaction to obtain a liposome mixture; S13. After the liposome mixture was cooled to room temperature, the Poria extract was slowly added to the liposome mixture, stirred evenly, and 0.5 to 1M sodium hydroxide was added dropwise to adjust the pH to 6 to 7 to obtain a first mixed solution; S14. Add the first mixed solution and 75% to 80% ethanol solution to a rotary evaporator and evaporate until membrane lipids are formed on the inner wall of the bottle on the rotary evaporator, continue to add 75% to 80% ethanol solution, and perform ultrasonic treatment to form an emulsion, then place the bottle on the rotary evaporator in a 30 to 40° C. water bath to remove the emulsion gel on the bottle, thereby obtaining a first liposome; S15. The pH of the first liposome was adjusted to 6-7 with phosphate buffer, and then a high-pressure homogenizer was used to obtain 200-220 nm Poria liposomes, which were freeze-dried at -4--10°C for 8-12 hours to obtain modified Poria polysaccharide.

[0006] Furthermore, the complex vitamin mainly comprises vitamin A, vitamin D and vitamin E, wherein the mass ratio of vitamin A, vitamin D and vitamin E is 1:(1~2):(1~3).

[0007] Furthermore, the mass ratio of the agave extract to acetic acid is 1:(2-4).

[0008] Furthermore, the preparation method of the agave extract comprises: S21. Take 80-90 g of dried fresh leaves and rhizomes of agave and crush them, pass them through a 50-60 mesh sieve, and then put them into a wall breaking machine to crush them to obtain an agave mixture; S22. Soaking the agave mixture in a 95-98% acetone solution for 1-3 hours, filtering to obtain an agave soaking solution, and maintaining the heating temperature at 40-50° C. to obtain a first mixture; S23. The first mixture was distilled with 95 to 98% ethanol for 1 to 3 hours, and then the first mixture after distillation was extracted with 2 to 5 ml of ethanol. After extraction 3 to 5 times, an agave extract was obtained; S24. The agave extract is concentrated by a rotary evaporator to 1 / 2 to 1 / 4 of the original volume to obtain an agave concentrate; S25. Subsequently, the mixture is centrifuged at 3000-4000 rpm for 3-5 min, and 0.5-1.0 M sodium hydroxide is added to the supernatant to adjust the pH to 6-7 to obtain an agave extract.

[0009] Furthermore, the composite probiotics mainly include Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium, and the weight ratio of Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium is (10-20): (5-7): (10-15).

[0010] Furthermore, the mass ratio of the composite probiotics, composite vitamins and limonin is (1-20):(2-4):(1-3).

[0011] On the other hand, based on the same inventive concept, the present invention also provides a method for preparing a feed additive containing a composite probiotic, which is applied to the feed additive containing a composite probiotic, comprising the following steps: S31. Add 100 parts of corn and 30 parts of wheat bran in a container, wash and drain, install it and crush it into slag, stir repeatedly and add 100 ~ 200g of pure water to obtain a first mixture; S32. The composite probiotics were inoculated into a liquid culture medium, cultured at 30-37 ° C for 24-36 h, and then the cells were collected by centrifugation and washed with sterile saline to prepare a composite probiotic suspension; S33. Add the composite probiotic suspension, agave extract, acetic acid, β-amylase, multivitamins, limonin, and taurine to the first mixture in a stirring state, and stir for 4 to 6 hours under heating conditions at a heating temperature of 30 to 35 ° C until fully mixed to obtain a second mixture; S34. The second mixture is compressed and granulated by a flat die granulator to form granular feed, which is packaged after cooling.

[0012] The mechanism of action of the above raw material components is as follows: Modified Poria cocos polysaccharide is formed by encapsulating Poria cocos polysaccharide with liposomes. Poria cocos polysaccharide comes from the fruiting body, mycelium and fermentation liquid of Poria cocos. Poria cocos is the dried sclerotium of the fungus Poria cocos of the genus Poria of the Polyporaceae family, and is an important medicinal and edible fungus. The double-layer structure of liposomes can effectively encapsulate Poria cocos polysaccharide, protect its properties from the influence of the external environment, and improve its stability. Modified Poria cocos polysaccharide can promote the growth of beneficial bacteria, such as Bifidobacterium and Lactobacillus acidophilus, while inhibiting the growth of harmful bacteria, such as Escherichia coli and Salmonella, thereby maintaining the balance of intestinal microecology. Modified Poria cocos polysaccharide can promote the growth and repair of intestinal mucosa, enhance the intestinal barrier function, reduce the intestinal absorption of harmful substances, and reduce the risk of intestinal inflammation.

[0013] Agave extract is obtained from the stems and leaves of Agave americana through a distillation extraction process. Agave extract is rich in a variety of active ingredients, such as polysaccharides, saponins, phenolic compounds, etc., which make it have good biocompatibility and stability. Among them, agave fructan is a water-soluble dietary fiber, mainly composed of short-chain fructose molecules, which are connected by (2-1) glycosidic bonds and have glucose units as the end groups of the chain. The active ingredients in agave extract can regulate the structure of intestinal flora, promote the growth of beneficial bacteria, and inhibit the reproduction of harmful bacteria. By reducing the content of ammonia in the intestine, maintaining the acid-base balance of the intestine, and providing suitable pH conditions for digestive enzymes to exert their highest activity, the digestion and absorption of nutrients are promoted.

[0014] The compound probiotics are mainly composed of Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium. Lactobacillus rhamnosus can strengthen the intestinal barrier, regulate immune response, inhibit the growth of harmful bacteria, improve digestive function, and reduce allergic reactions. Lactobacillus acidophilus lowers the intestinal pH value by producing lactic acid, inhibits harmful bacteria, and synthesizes B vitamins and vitamin K, enhances immunity, and lowers cholesterol. Bifidobacterium helps maintain the balance of intestinal microecology, promotes intestinal peristalsis, produces short-chain fatty acids with anti-inflammatory effects, strengthens the intestinal barrier, and lowers cholesterol and blood pressure.

[0015] As one of the short-chain fatty acids, acetic acid can lower the pH value of the intestine, provide a suitable living environment for beneficial bacteria, inhibit the growth of harmful bacteria, and thus maintain the balance of intestinal flora. At the same time, it promotes intestinal peristalsis, helps digest food and promotes defecation, and improves gastrointestinal motility and digestion and absorption functions. Acetic acid can also serve as the main energy source for intestinal cells, regulate the absorption of various nutrients and hormone production in the intestine, and participate in the energy metabolism process.

[0016] Limonin is a bitter compound found in citrus fruits and belongs to the tetracyclic triterpene lactones in plants. It is a fat-soluble compound with low solubility in water. Limonin is relatively stable at high temperatures, which makes it difficult to remove by heating. Limonin can stimulate intestinal smooth muscle and enhance intestinal motility, thereby helping food and digestive waste pass through the intestines more smoothly. At the same time, it can stimulate the gastrointestinal tract to secrete digestive juices, including gastric juice and intestinal juice, and improve digestion efficiency. Limonin can promote the growth of lactobacilli and bifidobacteria, help digestion, produce short-chain fatty acids (SCFAs), and regulate immune responses.

[0017] Corn and wheat bran are the main energy sources in feed. Corn has high starch and fat content, so it has high energy value and provides animals with a rich source of energy. Wheat bran is rich in crude fiber, which helps to increase the mechanical stimulation of the digestive tract of ruminants and promote digestion and absorption. The fiber components in wheat bran help promote intestinal peristalsis and improve intestinal health. β-amylase is an exoenzyme that can hydrolyze the non-reducing ends of starch molecules and cut off maltose molecules in turn. β-amylase mainly hydrolyzes α-1,4-glycosidic bonds to produce maltose as a product. Since β-amylase cannot hydrolyze the α-1,6-glycosidic bond in amylopectin, its hydrolysis stops when it encounters this bond. This characteristic allows β-amylase to break down starch in a targeted manner in feed and provide animals with easily absorbed sugars.

[0018] Taurine, also known as α-aminoethanesulfonic acid or β-aminoethanesulfonic acid, is a sulfur-containing non-protein amino acid. It can significantly improve the length, weight, villus height and crypt depth of the intestine. Taurine contributes to the normal development of the intestine, which is essential for the bone development, muscle growth and fur health of animals. Vitamins A, D and E in multivitamins have a protective effect on the intestinal mucosa, can enhance the intestinal barrier function, prevent harmful substances from entering the body, and promote the proliferation and differentiation of intestinal cells, which helps repair and regenerate intestinal tissues. By increasing the content of intestinal protein and DNA and reducing the apoptosis of intestinal epithelial cells, these vitamins participate in various metabolic processes in the body, promote protein synthesis, and accelerate cell division, thereby accelerating the growth rate of animals and improving body shape and meat quality.

[0019] In the composite probiotic feed additive, agave extract, modified Poria cocos polysaccharide and composite probiotics play a role through synergistic effects. During the preparation of feed additives, functional groups such as amino and hydroxyl groups in the molecular structure of modified Poria cocos polysaccharide interact with nutrients such as proteins, amino acids, polysaccharides in composite probiotics, and the composite probiotics and other nutrients are tightly wrapped in the complex formed by modified Poria cocos polysaccharide through physical adsorption and chemical bonding. This wrapping effect not only effectively prevents the loss of nutrients such as composite probiotics before entering the animal, but also enables the composite probiotics to be more fully absorbed and utilized by the animal body when consumed. When the complex enters the small intestine of the animal, the modified Poria cocos polysaccharide formed on its surface is decomposed, and the composite probiotics are exposed to the intestinal environment. Among them, probiotics such as Lactobacillus rhamnosus can activate receptors on the surface of intestinal epithelial cells by secreting specific short-chain fats as signals in the intestinal environment, thereby initiating cell signal transduction. In this process, agave extract further stimulates compound probiotics to secrete short-chain fats. Compound probiotics such as Lactobacillus rhamnosus can use agave extract for fermentation metabolism, breaking down carbohydrates into smaller molecules and producing a series of metabolites, further increasing the content of short-chain fats. The increase in short-chain fat content will affect the increase in signal transduction, and these signal transduction pathways can promote the enhancement of tight junctions, reduce intestinal permeability, and prevent harmful substances and pathogens from entering the body.

[0020] (3) Beneficial effects Compared with the prior art, the present invention has the following beneficial effects: 1. Modified tuckahoe polysaccharide is used as the core wrapping material to tightly wrap the compound probiotics, agave extract and other nutrients in the compound, and the modified tuckahoe polysaccharide film serves as the outer protective barrier to enhance the stability and antioxidant capacity of the compound. This dynamic balance system not only maintains the content of probiotics in the feed, but also enables the probiotics to be effectively absorbed and utilized in animals; 2. Agave extract can provide nutrients for probiotics. After reaching the animal's intestines, agave extract can continuously stimulate the compound probiotics to secrete signal molecules, which not only promotes communication and collaboration between probiotics, but also enhances the intestinal barrier function and improves the intestinal channel, thereby effectively preventing the invasion of harmful substances and pathogens; 3. Agave extract and modified Poria cocos polysaccharide are both natural plant ingredients. They are not only rich in bioactive substances, but also have good biocompatibility and stability with compound probiotics. They help probiotics to establish dominant flora in the intestines, and can also effectively resist the invasion of harmful bacteria and maintain the microecological balance of the intestines; 4. Agave extract, modified Poria cocos polysaccharide and compound probiotics work synergistically to promote the improvement of intestinal permeability in animals, enhance the tightness of intestinal epithelial cells, maintain intestinal health and promote nutrient absorption. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a SEM image of the modified Pachymaran of Example 1 of the present invention; Figure 2 This is the SEM image of the agave extract of Example 1 of the present invention. DETAILED DESCRIPTION

[0022] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] The experimental equipment and preparations of the embodiments described below are as follows: electronic balance (Sartorius, Germany), electric constant temperature water bath (Jiangsu Kedao), magnetic stirrer (Shanghai Meiyingpu), ultrasonic instrument (Shanghai Yixin), high-speed centrifuge (Guangzhou Jidi), vacuum drying oven (Shanghai Jiecheng), rotary evaporator (Shanghai Yaote), scanning electron microscope (Zeiss, Germany), specific surface area analyzer (Beijing Best Instrument Technology), freeze dryer (Shanghai Pudong Freeze Drying), pH meter (Shanghai Yidian), constant temperature incubator (Shanghai Hetian), ultrasonic vibrator (ELMA- E5K); chemicals and reagents were purchased from Sigma-Aldrich.

[0024] Example 1: This example discloses a feed additive containing a composite probiotic, comprising the following raw materials in parts by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the weight ratio of the modified Poria cocos polysaccharide to the composite probiotics is 1:6. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15,000~20,000 Da.

[0025] In the composite probiotic feed additive, agave extract, modified Poria cocos polysaccharide and composite probiotics play a synergistic role. Table 1 shows the SEM of modified Poria cocos polysaccharide. It can be seen from the figure that modified Poria cocos polysaccharide presents a sheet structure and has a hollow structure, which is easy to encapsulate nutrients. Modified Poria cocos polysaccharide can encapsulate the composite probiotics and agave extract in the feed to form a stable complex. Figure 2The SEM of agave extract is a distinct spherical structure with good stability. It not only ensures the stability and integrity of these active ingredients during feed processing, storage and animal digestion. By reducing the damage of the external environment to the compound probiotics and agave extract, the modified Poria cocos polysaccharide significantly reduces the loss rate of these beneficial substances, thereby improving their bioavailability. When these complexes enter the animal intestine, the agave extract further plays its unique role. It can promote the compound probiotics to secrete specific signal molecules, which act on intestinal epithelial cells and enhance the tight junctions of intestinal epithelial cells through a series of complex signal transduction pathways, thereby maintaining the integrity of the intestinal barrier. The signal molecules such as exopolysaccharides and bacteriocins secreted by the compound probiotics are accurately recognized by specific receptors of intestinal epithelial cells (such as pattern recognition receptors such as TLR), thereby activating a series of signal transduction pathways, including the activation of intracellular kinases (such as ERK1 / 2, p38 MAPK), the regulation of intracellular calcium signals, and the activation of transcription factors (such as NF-κB, STATs). These activated transcription factors then induce the expression of tight junction protein-related genes (such as occludin and claudin), prompting cells to synthesize and transport these key proteins to the cell membrane. On the cell membrane, proteins such as occludin and claudin bind tightly to the corresponding proteins of adjacent cells to form and strengthen tight junctions, thereby strictly limiting the permeability of substances between cells and significantly enhancing the intestinal barrier function. Taking Lactobacillus rhamnosus as an example, the signal molecules it secretes are mainly short-chain fatty acids (acetic acid, propionic acid, butyric acid, etc.) and bacterial metabolites (indole, etc.). When it enters the intestinal cells, the short-chain fatty acids (such as butyric acid) secreted can activate GPR41, GPR43 and GPR109A receptors. Agave extract can further provide energy to stimulate it to secrete more short-chain fatty acids, thereby triggering downstream signaling pathways, which can further promote the expression of tight junction proteins (such as ZO-1, occludin and claudin) and enhance the intestinal barrier function. This process not only maintains the integrity of the intestinal barrier, but also promotes the timely repair of the barrier function. This role is crucial for resisting pathogen invasion, reducing intestinal inflammation and improving the balance of intestinal microecology. In this process, agave extract is rich in a variety of nutrients that can be used by microorganisms, which can provide the necessary energy source for compound probiotics and support their survival, reproduction and metabolic activities in the intestine. In addition, agave extract can also promote the nutrient absorption function of intestinal epithelial cells. By regulating the metabolic activity and gene expression of intestinal epithelial cells, it helps to increase the absorption efficiency of nutrients, improve the utilization rate of feed, and thus promote the growth and development of animals.

[0026] The preparation method of the modified tuckahoe polysaccharide comprises: S11. Add 50-60 g of Poria cocos to 75%-80% ethanol solution under heating and stirring, heat to 60-70°C, stir for 4-5 hours, stir evenly until completely dissolved, and obtain Poria cocos extract; S12. 5 to 6 g of phosphatidylethanolamine was stirred at room temperature, and then a 75% to 80% ethanol solution was added to promote the reaction to obtain a liposome mixture; S13. After the liposome mixture was cooled to room temperature, the Poria extract was slowly added to the liposome mixture, stirred evenly, and 0.5 to 1M sodium hydroxide was added dropwise to adjust the pH to 6 to 7 to obtain a first mixed solution; S14. Add the first mixed solution and 75% to 80% ethanol solution to a rotary evaporator and evaporate until membrane lipids are formed on the inner wall of the bottle on the rotary evaporator, continue to add 75% to 80% ethanol solution, and perform ultrasonic treatment to form an emulsion, then place the bottle on the rotary evaporator in a 30 to 40° C. water bath to remove the emulsion gel on the bottle, thereby obtaining a first liposome; S15. The pH of the first liposome was adjusted to 6-7 with phosphate buffer, and then a high-pressure homogenizer was used to obtain 200-220 nm Poria liposomes, which were freeze-dried at -4--10°C for 8-12 hours to obtain modified Poria polysaccharide.

[0027] The complex vitamin mainly comprises vitamin A, vitamin D and vitamin E, wherein the mass ratio of vitamin A, vitamin D and vitamin E is 1:1:1.

[0028] The mass ratio of the corn to the β-amylase is 1:0.5.

[0029] The mass ratio of the agave extract to acetic acid is 1:2.

[0030] The preparation method of the composite probiotics comprises: S21. Take 80-90 g of dried fresh leaves and rhizomes of agave and crush them, pass them through a 50-60 mesh sieve, and then put them into a wall breaking machine to crush them to obtain an agave mixture; S22. Soaking the agave mixture in a 95-98% acetone solution for 1-3 hours, filtering to obtain an agave soaking solution, and maintaining the heating temperature at 40-50° C. to obtain a first mixture; S23. The first mixture was distilled with 95 to 98% ethanol for 1 to 3 hours, and then the first mixture after distillation was extracted with 2 to 5 ml of ethanol. After extraction 3 to 5 times, an agave extract was obtained; S24. The agave extract is concentrated by a rotary evaporator to 1 / 2 to 1 / 4 of the original volume to obtain an agave concentrate; S25. Subsequently, the mixture is centrifuged at 3000-4000 rpm for 3-5 min, and 0.5-1.0 M sodium hydroxide is added to the supernatant to adjust the pH to 6-7 to obtain an agave extract.

[0031] The composite probiotics mainly include Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium, and the weight ratio of Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium is 10:5:12.

[0032] A method for preparing a composite probiotics-containing feed additive, applied to the composite probiotics-containing feed additive, comprises the following steps: S31. Add 100 parts of corn and 30 parts of wheat bran in a container, wash and drain, install it and crush it into slag, stir repeatedly and add 100 ~ 200g of pure water to obtain a first mixture; S32. The composite probiotics were inoculated into a liquid culture medium, cultured at 30-37 ° C for 24-36 h, and then the cells were collected by centrifugation and washed with sterile saline to prepare a composite probiotic suspension; S33. Add the composite probiotic suspension, agave extract, acetic acid, β-amylase, multivitamins, limonin, and taurine to the first mixture in a stirring state, and stir for 4 to 6 hours under heating conditions at a heating temperature of 30 to 35 ° C until fully mixed to obtain a second mixture; S34. The second mixture is compressed and granulated by a flat die granulator to form granular feed, which is packaged after cooling.

[0033] Example 2: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 10 parts of agave extract, 10 parts of acetic acid, 2 parts of β-amylase, 30 parts of composite probiotics, 150 parts of corn, and 25 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:5. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0034] Example 3: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 10 parts of agave extract, 15 parts of acetic acid, 3 parts of β-amylase, 40 parts of composite probiotics, 200 parts of corn, and 30 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:4. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0035] Example 4: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 15 parts of agave extract, 15 parts of acetic acid, 3 parts of β-amylase, 40 parts of composite probiotics, 200 parts of corn, and 30 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:6. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0036] Example 5: This example discloses a feed additive containing a composite probiotic, including the following raw materials by weight: 20 parts of agave extract, 15 parts of acetic acid, 3 parts of β-amylase, 40 parts of composite probiotics, 200 parts of corn, and 30 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:5. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing a composite probiotic in this example is consistent with that in Example 1.

[0037] Example 6: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:5. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0038] Example 6: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:4. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0039] Example 7: This example discloses a feed additive containing composite probiotics, including the following raw materials by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:3. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this example is consistent with that in Example 1. The preparation method of a feed additive containing composite probiotics in this example is consistent with that in Example 1.

[0040] Control group 1: The difference between this embodiment and embodiment 1 is that modified Poria cocos polysaccharide is not contained. This embodiment discloses a feed additive containing composite probiotics, comprising the following raw materials in parts by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The preparation method of the composite probiotics in this embodiment is consistent with that in embodiment 1. The preparation method of a feed additive containing composite probiotics in this embodiment is consistent with that in embodiment 1.

[0041] Control group 2: The difference between this embodiment and embodiment 1 is that it does not contain composite probiotics. This embodiment discloses a feed additive containing composite probiotics, comprising the following raw materials in parts by weight: 10 parts of agave extract, 5 parts of acetic acid, 1 part of β-amylase, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, which is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000Da. The preparation method of the modified Poria cocos polysaccharide in this embodiment is consistent with that in embodiment 1. The preparation method of a feed additive containing composite probiotics in this embodiment is consistent with that in embodiment 1.

[0042] Control group 3: The difference between this embodiment and embodiment 1 is that agave extract is not contained. This embodiment discloses a feed additive containing composite probiotics, comprising the following raw materials in parts by weight: 5 parts of acetic acid, 1 part of β-amylase, 20 parts of composite probiotics, 100 parts of corn, and 20 parts of wheat bran. The composite probiotic feed additive also includes modified Poria cocos polysaccharide, and the modified Poria cocos polysaccharide and the composite probiotics are in a weight ratio of 1:3. The modified Poria cocos polysaccharide is prepared by encapsulating Poria cocos polysaccharide in a lipid bilayer structure, and the molecular weight of the modified Poria cocos polysaccharide is 15000~20000 Da. The preparation method of the modified Poria cocos polysaccharide and the composite probiotics in this embodiment is consistent with that in embodiment 1. The preparation method of a feed additive containing composite probiotics in this embodiment is consistent with that in embodiment 1.

[0043] Control group 4: This embodiment differs from embodiment 1 in that it does not contain agave extract, modified tuckahoe polysaccharide and composite probiotics. This embodiment discloses a feed additive containing composite probiotics, comprising the following raw materials in parts by weight: 5 parts of acetic acid, 1 part of β-amylase, 100 parts of corn, and 20 parts of wheat bran. The preparation method of a feed additive containing composite probiotics in this embodiment is consistent with that in embodiment 1.

[0044] Blank group 1: No compound probiotic feed additive was added to the feed.

[0045] Effect evaluation: Dutch rabbit growth performance evaluation test: 50 young Dutch rabbits (1-2 months) weighing 0.9-1.0 kg were selected, half male and half female, and randomly divided into 5 groups, 10 in each group. Experimental environment: temperature 18-22℃, humidity 40%-50%, free drinking water, ensure air circulation, and feed at 12:00 am and 6:00 pm every day. Add 50ml of compound probiotic feed additive to every 10 catties of Dutch rabbit feed in each experimental group, mix well and feed, add the same amount of mineral water to the blank group and mix well. The experimental period is six weeks. Every seven days in a quiet and undisturbed environment, gently place the Dutch rabbit on the scale. Make sure it stands stably, do not jump or move, and weigh and record. The effect of compound probiotic feed additive on the growth of Dutch rabbits was evaluated by calculating the weight gain and feed conversion rate of Dutch rabbits. The weight of Dutch rabbits was weighed every seven days in the experiment and the average value of each group was calculated.

[0046] Feed conversion ratio (FCR) = feed intake / weight gain Table 1 shows the statistical results of the growth of Dutch rabbits in each experimental group.

[0047] Table 1 is the statistical results of the growth of each experimental group feed. It can be seen from Table 1 that after the composite probiotic feed additive prepared by each experimental group was added to the feed, there was a significant difference in the weight of the Dutch rabbits. The control group showed a higher FCR ratio. The higher the FCR, the lower the feed conversion rate, that is, the animal uses more feed to obtain less weight gain, and the feeding efficiency is also lower. Therefore, the lower the FCR, the better the effect of the feed. Comparing the weight gain of Dutch rabbits in Examples 1 to 7 with that in Control Groups 1 to 4, it can be found that, overall, the composite probiotic feed additive prepared in Examples 1 to 7 will make the weight gain of Dutch rabbits more obvious, and the FCR ratio is 1.21 kg / kg, which is lower than that of other experimental groups, control groups and blank groups. This shows that the feed effect of the composite probiotic feed additive prepared in Example 1 is the best, and the weight gain of animals is more obvious, while the FCR prepared in Control Groups 1 to 3 is <1.21 kg / kg, which is at an ordinary level, which shows that by adding agave extract, modified tuckahoe polysaccharide and composite probiotics at the same time during the preparation of the feed additive, an excellent composite probiotic feed additive can be obtained. After the above limited experiments, the application effect of a composite probiotic feed additive containing an embodiment of the present invention 1 is significant. By adding agave extract, modified tuckahoe polysaccharide and composite probiotics at the same time during the production process, the utilization rate of feed can be significantly improved, and a multifunctional feed additive is provided for animal health. The three form a dynamic balance system to jointly maintain a healthy and nutritious feed additive.

[0048] Finally, it should be noted that: although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A feed additive containing composite probiotics, characterized in that: The invention comprises the following raw materials in parts by weight: 10-20 parts of agave extract, 5-15 parts of acetic acid, 1-3 parts of beta-amylase, 20-40 parts of compound probiotics, 20-40 parts of compound vitamins, 5-15 parts of limonin, 10-20 parts of taurine, 100-200 parts of corn, and 20-30 parts of wheat bran; The composite probiotic feed additive also includes: Modified pachymaran; The modified pachymaran and the composite probiotics are in a weight ratio of 1:(3-6); The modified pachymaran is prepared by encapsulating pachymaran in a lipid bilayer structure, and the molecular weight of the modified pachymaran is 15000-20000 Da.

2. A composite probiotic feed additive according to claim 1, characterized in that: The preparation method of the modified tuckahoe polysaccharide comprises: S11. Add 50-60 g of Poria cocos to 75%-80% ethanol solution under heating and stirring, heat to 60-70°C, stir for 4-5 hours, stir evenly until completely dissolved, and obtain Poria cocos extract; S12. 5 to 6 g of phosphatidylethanolamine was stirred at room temperature, and then a 75% to 80% ethanol solution was added to promote the reaction to obtain a liposome mixture; S13. After the liposome mixture was cooled to room temperature, the Poria extract was slowly added to the liposome mixture, stirred evenly, and 0.5 to 1M sodium hydroxide was added dropwise to adjust the pH to 6 to 7 to obtain a first mixed solution; S14. Add the first mixed solution and 75% to 80% ethanol solution to a rotary evaporator and evaporate until membrane lipids are formed on the inner wall of the bottle on the rotary evaporator, continue to add 75% to 80% ethanol solution, and perform ultrasonic treatment to form an emulsion, then place the bottle on the rotary evaporator in a 30 to 40° C. water bath to remove the emulsion gel on the bottle, thereby obtaining a first liposome; S15. The pH of the first liposome was adjusted to 6-7 with phosphate buffer, and then a high-pressure homogenizer was used to obtain 200-220 nm Poria liposomes, which were freeze-dried at -4--10°C for 8-12 hours to obtain modified Poria polysaccharide.

3. The composite probiotic feed additive according to claim 1, characterized in that: The complex vitamin mainly comprises vitamin A, vitamin D and vitamin E, wherein the mass ratio of vitamin A, vitamin D and vitamin E is 1:(1~2):(1~3).

4. The composite probiotic feed additive according to claim 1, characterized in that: The mass ratio of the agave extract to acetic acid is 1:(2-4).

5. The composite probiotic feed additive according to claim 1, characterized in that: The preparation method of the agave extract comprises: S21. Take 80-90 g of dried agave leaves and rhizomes, crush them, pass them through a 50-60 mesh sieve, and then put them into a wall breaking machine to crush them to obtain an agave mixture; S22. Soaking the agave mixture in a 95-98% acetone solution for 1-3 hours, filtering to obtain an agave soaking solution, and maintaining the heating temperature at 40-50° C. to obtain a first mixture; S23. The first mixture was distilled with 95 to 98% ethanol for 1 to 3 hours, and then the first mixture after distillation was extracted with 2 to 5 ml of ethanol. After extraction 3 to 5 times, an agave extract was obtained; S24. The agave extract is concentrated by a rotary evaporator to 1 / 2 to 1 / 4 of the original volume to obtain an agave concentrate; S25. Subsequently, the mixture is centrifuged at 3000-4000 rpm for 3-5 min, and 0.5-1.0 M sodium hydroxide is added to the supernatant to adjust the pH to 6-7 to obtain an agave extract.

6. The composite probiotic feed additive according to claim 1, characterized in that: The composite probiotics mainly include Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium, and the weight ratio of Lactobacillus rhamnosus, Lactobacillus acidophilus and Bifidobacterium is (10-20): (5-7): (10-15).

7. The composite probiotic feed additive according to claim 1, characterized in that: The mass ratio of the composite probiotics, composite vitamins and limonin is (1-20):(2-4):(1-3).

8. A method for preparing a composite probiotic feed additive, which is used to prepare a composite probiotic feed additive as claimed in any one of claims 1 to 7, characterized in that: The method comprises the following steps: S31. Add 100 parts of corn and 30 parts of wheat bran in a container, wash and drain, install it and crush it into slag, stir repeatedly and add 100 ~ 200g of pure water to obtain a first mixture; S32. The composite probiotics were inoculated into a liquid culture medium, cultured at 30-37 ° C for 24-36 h, and then the cells were collected by centrifugation and washed with sterile saline to prepare a composite probiotic suspension; S33. Add the composite probiotic suspension, agave extract, acetic acid, β-amylase, multivitamins, limonin, and taurine to the first mixture in a stirring state, and stir for 4 to 6 hours under heating conditions at a heating temperature of 30 to 35 ° C until fully mixed to obtain a second mixture; S34. The second mixture is compressed and granulated by a flat die granulator to form a granular feed, which is packaged after cooling to obtain a composite probiotic feed additive.

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