Preparation method of fattening and growth-promoting feed additive for ruminant animals

Abstract

The invention relates to the technical field of feed additives, and discloses a ruminant animal fattening and growth-promoting feed additive which comprises the following components in percentage by mass: 22-28 parts of phytosterol, 42-48 parts of a rumen protection type amino acid compound, 11-14 parts of a compound probiotic preparation, 6-7.5 parts of a compound enzyme preparation, 9-11 parts of a trace element compound and 4-4.8 parts of a buffer system. According to the preparation method of the fattening and growth-promoting feed additive for ruminant animals, a modified hydrogenated palm oil and lecithin compound is used as a rumen protection type amino acid coating material, and a high-pressure homogenizer is used for performing multi-stage homogenizing treatment, so that a coating layer is more compact in structure, and the weight of the coating layer is increased; the rumen microorganism lipase decomposition resistance is enhanced through the synergistic effect of the composite material, particle distribution is optimized in combination with a refined homogenizing process, the release controllability and absorption efficiency of amino acid in small intestines are improved, and the problems that a traditional single coating material is easy to decompose and the particle size is extensively controlled are solved.

Description

Technical Field

[0001] The invention relates to the technical field of feed additives, in particular to a method for preparing a feed additive for fattening and promoting growth of ruminant animals. Background Art

[0002] Ruminant animal fattening and growth promoting feed additives are a type of special preparation that is added to ruminant feed through a scientific ratio of multiple functional ingredients to promote their growth and development, increase feed conversion efficiency and improve their health status. Its core function is to improve animal growth performance by regulating the rumen microecological environment, optimizing the absorption and utilization pathways of nutrients and maintaining the body's metabolic balance.

[0003] However, the existing ruminant fattening and growth-promoting feed additives still have significant technical bottlenecks in formula design and preparation technology; on the one hand, traditional rumen-protected amino acids mostly use a single hydrogenated oil as the coating material, and the coating layer is easily decomposed by microbial lipase in the complex environment of the rumen, resulting in low absorption efficiency of amino acids in the small intestine, and the particle size control in the homogenization process in the coating process is rough, affecting the stability of the ingredients; on the other hand, probiotic preparations generally have the problem of a single strain combination, lack of strains with both high temperature resistance and efficient colonization ability, and the freeze-dried protective agent formula and drying process parameters are not optimized for the rumen environment of ruminants, resulting in a low survival rate of live bacteria after granulation, making it difficult to form a sustained and effective flora regulation effect in the animal body. Therefore, a preparation method for a ruminant animal fattening and growth-promoting feed additive is proposed. Summary of the Invention

[0004] (1) Technical problems solved

[0005] In response to the shortcomings of the existing technology, the present invention provides a method for preparing a feed additive for fattening and promoting growth of ruminant animals, which has the advantages of improving the efficiency and stability of amino acid absorption and enhancing the stress resistance of probiotics. It solves the problems in existing additives that rumen-protected amino acids are easily decomposed, and probiotics are single in species and have insufficient colonization ability.

[0006] (2) Technical solution

[0007] To achieve the above-mentioned purpose of improving amino acid absorption efficiency and stability and enhancing probiotic resistance, the present invention provides the following technical solution: a feed additive for fattening and promoting growth of ruminant animals, comprising the following components and their mass percentages: 22-28 parts of phytosterols, 42-48 parts of rumen-protected amino acid complex, 11-14 parts of a composite probiotic preparation, 6-7.5 parts of a composite enzyme preparation, 9-11 parts of a trace element complex, and 4-4.8 parts of a buffer system;

[0008] The β-sitosterol content in the phytosterol is not less than 65%, and the stigmasterol content is 15-20%;

[0009] The rumen-protected amino acid complex is composed of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.2-2.8:1, wherein the coating material of the rumen-protected lysine is a complex of modified hydrogenated palm oil with a hydroxyl content of 15-20 mol% and lecithin, and the amount of the coating material is 22-24% of the weight of the lysine;

[0010] The composite probiotic preparation is composed of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum and Bacillus coagulans, and the ratio of viable bacteria count is 3.2-3.8:2.1-2.9:1:0.8-1.2;

[0011] The complex enzyme preparation comprises cellulase with an enzyme activity of 35,000-45,000 U / g, xylanase with an enzyme activity of 20,000-30,000 U / g, and amylase with an enzyme activity ratio of 10,000-15,000 U / g, with the ratio of the three enzymes being 3.3-3.7:2.2-2.8:1;

[0012] The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, and the weight ratio of each component is 5.2-5.8:1.1-1.9:3.1-3.9:1.1-1.9:0.3-0.7;

[0013] The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, and the weight ratio of the three is 2.1-2.9:1.2-1.8:0.5-0.9.

[0014] Preferably, the phytosterols are separated and prepared by silica gel column chromatography, using n-hexane-ethyl acetate with a volume ratio of 8:2 as the eluent, collecting the β-sitosterol elution peak with a retention time of 28-32 minutes, and the purity after freeze crystallization is not less than 98%;

[0015] The stigmasterol is extracted by supercritical CO2 at an extraction pressure of 25-30 MPa and a temperature of 45-50° C., with a yield of not less than 92%. β-sitosterol and stigmasterol are compounded at a weight ratio of 3.5-4.5:1.

[0016] Preferably, the cellulase is derived from a mutant strain of Trichoderma reesei QM6a, and is purified by an ammonium sulfate salting-out method after liquid fermentation, with a salting-out saturation of 30-50%, an enzyme protein purity of not less than 90%, and an enzyme activity retention rate of not less than 95% at an optimum pH of 5.8-6.2;

[0017] The xylanase is introduced with histidine at the 125th amino acid by using site-directed mutagenesis technology, so that its optimum action temperature is increased to 50-55° C., and the residual enzyme activity is not less than 70% after being treated at 60° C. for 30 minutes.

[0018] A method for preparing a feed additive for fattening and promoting growth of ruminant animals comprises the following steps:

[0019] S1. Preparation of protective coating:

[0020] Modified hydrogenated palm oil having a hydroxyl content of 15-20 mol% and lecithin are mixed in a weight ratio of 4.5-5.5:1, heated to 65-75° C. to melt, lysine raw material is added so that the coating material accounts for 22-24% of the weight of the lysine, and the mixture is stirred at a speed of 200-300 r / min for 15-25 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 120-140 MPa to prepare rumen-protected lysine.

[0021] The same coating process is used to prepare rumen-protected methionine, with the coating material dosage being 18-22% of the weight of methionine;

[0022] S2. Preparation of probiotic preparations:

[0023] Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium, modified brain heart infusion medium, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 180-220 rpm for 20-22 h. The culture broth was centrifuged at 5500-6500 rpm for 12-18 min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 12-14% of the cell weight, wherein the protective agent consisted of trehalose and skim milk powder in a weight ratio of 1.2:1, and freeze-dried at -55-45°C and a vacuum condition of 1-3 Pa for 30-34 h.

[0024] S3. Microencapsulation treatment:

[0025] The phytosterols are mixed with a wall material, wherein the wall material is composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5, and the weight ratio of the phytosterols to the wall material is 1:3.2-3.8, to prepare an aqueous solution with a solid content of 16-19%, stirring at 52-58° C. and a stirring speed of 250-350 r / min for 35-45 minutes, and spray drying to obtain microcapsules. During the spray drying process, the air inlet temperature is controlled at 185-195° C., and the air outlet temperature is controlled at 82-88° C.

[0026] S4, gradient mixing process:

[0027] Primary mixing: Add the rumen-protected amino acid complex, complex enzyme preparation, and buffer system into a twin-shaft paddle mixer and mix at a speed of 120-140 r / min for 12-18 minutes;

[0028] Secondary mixing: Add probiotic preparation and phytosterol microcapsules, switch to planetary mixing mode, control the revolution speed to 80-100 r / min and the rotation speed to 200-240 r / min, and mix for 22-28 minutes;

[0029] Three-stage mixing: prepare the trace element complex into an aqueous solution with a concentration of 0.6-0.9 g / mL, spray it evenly into the mixer through an atomizing nozzle at a pressure of 0.4-0.6 MPa, and continue mixing for 10-15 minutes;

[0030] S5. Temperature controlled granulation:

[0031] The mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 62-68°C, the screw speed is 180-220r / min, the granulation pressure is 0.35-0.45MPa, and the obtained granules are sieved through an 85-95 mesh sieve. The sieve material is crushed and then mixed with the new mixture in a ratio of 1:3 and granulated again.

[0032] Preferably, 0.15-0.25 wt% of cysteine ​​hydrochloride is added to the modified MRS medium to maintain the integrity of Lactobacillus plantarum cells through sulfhydryl groups, thereby increasing the survival rate of bacteria in the logarithmic growth phase by 20%;

[0033] The modified brain heart infusion medium is added with 0.25-0.35 wt% of ox bile salts to induce Enterococcus faecalis to produce bile salt hydrolase, thereby enhancing its colonization ability in the high bile salt environment of the rumen and increasing the bile salt tolerance of Enterococcus faecalis to 0.5 wt%.

[0034] Preferably, the spray drying process uses a pressure atomizer with a speed of 20000±2000r / min and an inlet air humidity of 15-20%, so that the average particle size of the microcapsules is controlled at 10-15μm. Scanning electron microscopy shows that the surface porosity of the microcapsules does not exceed 5% and the specific surface area is not less than 50m 2 / g;

[0035] The contact time between the atomized droplets and the hot air is controlled at 0.8-1.2 seconds. The DVS dynamic water vapor adsorption instrument tests that the moisture absorption rate of the microcapsules does not exceed 8% when placed in an environment with a relative humidity of 75% for 24 hours.

[0036] Preferably, the high-pressure homogenizer adopts a two-stage homogenizing valve structure, the cone angle of the first-stage homogenizing valve is 60°±5°, and the cone angle of the second-stage homogenizing valve is 45°±5°. When monitored by a laser particle size analyzer, the D90 of the coated particles after homogenization is ≤12 μm, and the particle size distribution span is (D90-D10) / D50≤1.5;

[0037] During the homogenization process, the temperature of the feed liquid is controlled at 40-45°C by jacket circulating water to avoid thermal degradation of the lysine amino group. HPLC testing shows that the amino acid purity is not less than 99.2%.

[0038] Preferably, the particles are dried using a fluidized bed drying process with an air inlet temperature of 70-75° C. and a material fluidization velocity of 0.8-1.2 m / s. The moisture content of the particles after drying is not more than 7.5% as determined by Karl Fischer titration, and the surface is coated with 0.2-0.3 wt% of magnesium stearate, thereby improving moisture resistance by 30%.

[0039] The number of probiotics is detected by MPN-PCR method, wherein the spore rate of Bacillus coagulans is not less than 90%, and the viable bacteria survival rate after heat shock treatment at 65° C. for 10 minutes is not less than 85%, which meets the detection requirements of AOAC official analysis method 2015.12.

[0040] (3) Beneficial effects

[0041] Compared with the prior art, the present invention provides a method for preparing a feed additive for fattening and promoting growth of ruminant animals, which has the following beneficial effects:

[0042] 1. The preparation method of the feed additive for fattening and promoting growth of ruminant animals adopts a complex of modified hydrogenated palm oil and lecithin as a rumen-protected amino acid coating material, and uses a high-pressure homogenizer to perform multi-stage homogenization treatment to make the coating layer structure denser. The synergistic effect of the composite material enhances the decomposition ability of anti-rumen microbial lipase, and the refined homogenization process is combined to optimize the particle distribution, thereby improving the controllable release and absorption efficiency of amino acids in the small intestine, thereby solving the problems of easy decomposition and extensive particle size control of traditional single coating materials.

[0043] 2. The preparation method of the feed additive for fattening and promoting growth of ruminant animals comprises the following steps: forming a composite probiotic preparation of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum and Bacillus coagulans, adding cysteine ​​hydrochloride and ox bile salt to the improved culture medium to optimize the bacterial culture conditions, combining trehalose and skim milk powder composite protective agents and low-temperature vacuum freeze-drying process, improving the diversity of the bacterial community through the synergistic effect of multiple bacterial strains, enhancing the stress resistance of the bacterial body by using culture medium components, maintaining the activity of live bacteria with the help of protective agents and freeze-drying process, thereby solving the problems of single probiotic strain, insufficient high temperature resistance and rumen colonization ability. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 The present invention is a flow chart of the preparation method of the feed additive for fattening and promoting growth of ruminant animals. DETAILED DESCRIPTION

[0045] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the embodiments of the present invention and the accompanying drawings. 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 making creative efforts are within the scope of protection of the present invention.

[0046] See also Figure 1 A feed additive for fattening and promoting growth of ruminant animals, comprising the following components and their mass percentages: 22-28 parts of phytosterols, 42-48 parts of rumen-protected amino acid complex, 11-14 parts of a compound probiotic preparation, 6-7.5 parts of a compound enzyme preparation, 9-11 parts of a trace element complex, and 4-4.8 parts of a buffer system;

[0047] The β-sitosterol content in the phytosterol is not less than 65%, and the stigmasterol content is 15-20%;

[0048] The rumen-protected amino acid complex is composed of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.2-2.8:1, wherein the coating material of the rumen-protected lysine is a complex of modified hydrogenated palm oil with a hydroxyl content of 15-20 mol% and lecithin, and the amount of the coating material is 22-24% of the weight of the lysine;

[0049] The composite probiotic preparation is composed of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum and Bacillus coagulans, and the ratio of viable bacteria count is 3.2-3.8:2.1-2.9:1:0.8-1.2;

[0050] The complex enzyme preparation comprises cellulase with an enzyme activity of 35,000-45,000 U / g, xylanase with an enzyme activity of 20,000-30,000 U / g, and amylase with an enzyme activity ratio of 10,000-15,000 U / g, with the ratio of the three enzymes being 3.3-3.7:2.2-2.8:1;

[0051] The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, and the weight ratio of each component is 5.2-5.8:1.1-1.9:3.1-3.9:1.1-1.9:0.3-0.7;

[0052] The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, and the weight ratio of the three is 2.1-2.9:1.2-1.8:0.5-0.9.

[0053] A method for preparing a feed additive for fattening and promoting growth of ruminant animals comprises the following steps:

[0054] S1. Preparation of protective coating:

[0055] Modified hydrogenated palm oil having a hydroxyl content of 15-20 mol% and lecithin are mixed in a weight ratio of 4.5-5.5:1, heated to 65-75° C. to melt, lysine raw material is added so that the coating material accounts for 22-24% of the weight of the lysine, and the mixture is stirred at a speed of 200-300 r / min for 15-25 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 120-140 MPa to prepare rumen-protected lysine.

[0056] The same coating process is used to prepare rumen-protected methionine, with the coating material dosage being 18-22% of the weight of methionine;

[0057] S2. Preparation of probiotic preparations:

[0058] Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium, modified brain heart infusion medium, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 180-220 rpm for 20-22 h. The culture broth was centrifuged at 5500-6500 rpm for 12-18 min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 12-14% of the cell weight, wherein the protective agent consisted of trehalose and skim milk powder in a weight ratio of 1.2:1, and freeze-dried at -55-45°C and a vacuum condition of 1-3 Pa for 30-34 h.

[0059] S3. Microencapsulation treatment:

[0060] The phytosterols are mixed with a wall material, wherein the wall material is composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5, and the weight ratio of the phytosterols to the wall material is 1:3.2-3.8, to prepare an aqueous solution with a solid content of 16-19%, stirring at 52-58° C. and a stirring speed of 250-350 r / min for 35-45 minutes, and spray drying to obtain microcapsules. During the spray drying process, the air inlet temperature is controlled at 185-195° C., and the air outlet temperature is controlled at 82-88° C.

[0061] S4, gradient mixing process:

[0062] Primary mixing: Add the rumen-protected amino acid complex, complex enzyme preparation, and buffer system into a twin-shaft paddle mixer and mix at a speed of 120-140 r / min for 12-18 minutes;

[0063] Secondary mixing: Add probiotic preparation and phytosterol microcapsules, switch to planetary mixing mode, control the revolution speed to 80-100 r / min and the rotation speed to 200-240 r / min, and mix for 22-28 minutes;

[0064] Three-stage mixing: prepare the trace element complex into an aqueous solution with a concentration of 0.6-0.9 g / mL, spray it evenly into the mixer through an atomizing nozzle at a pressure of 0.4-0.6 MPa, and continue mixing for 10-15 minutes;

[0065] S5. Temperature controlled granulation:

[0066] The mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 62-68°C, the screw speed is 180-220r / min, the granulation pressure is 0.35-0.45MPa, and the obtained granules are sieved through an 85-95 mesh sieve. The sieve material is crushed and then mixed with the new mixture in a ratio of 1:3 and granulated again.

[0067] Example 1:

[0068] This embodiment provides a low-concentration formula suitable for young ruminants or the initial use stage of feed additives, which improves the rumen microecological environment through mild nutritional regulation and enhances the adaptability of young animals to feed.

[0069] A feed additive for fattening and promoting growth of ruminants, comprising the following components and their mass percentages:

[0070] 22 parts of plant sterols, 42 parts of rumen-protected amino acid complex, 11 parts of compound probiotic preparation, 6 parts of compound enzyme preparation, 9 parts of trace element complex, 4 parts of buffer system; of which:

[0071] The phytosterols contain 65% beta-sitosterol and 15% stigmasterol, and the two are compounded in a weight ratio of 3.5:1;

[0072] The rumen-protected amino acid complex consists of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.2:1. The coating material for the rumen-protected lysine is a complex of modified hydrogenated palm oil with a 15 mol% hydroxyl content and lecithin, with the coating material amounting to 22% of the weight of the lysine. The coating material for the rumen-protected methionine is 18% of the weight of the methionine.

[0073] The composite probiotic preparation consists of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans, with a live bacterial count ratio of 3.2:2.1:1:0.8;

[0074] The complex enzyme preparation contains 35,000 U / g cellulase, 20,000 U / g xylanase, and 10,000 U / g amylase, with the ratio of the three enzyme activities being 3.3:2.2:1.

[0075] The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, with the weight ratio of each component being 5.2:1.1:3.1:1.1:0.3;

[0076] The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, with a weight ratio of 2.1:1.2:0.5.

[0077] A method for preparing a feed additive for fattening and promoting growth of ruminant animals comprises the following steps:

[0078] Step 1: Preparing a protective coating: Modified hydrogenated palm oil with a hydroxyl content of 15 mol% and lecithin are mixed in a weight ratio of 4.5:1, heated to 65°C for melting, lysine raw material is added so that the coating material accounts for 22% of the weight of the lysine, and mixing is carried out at a stirring speed of 200 r / min for 15 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 120 MPa to prepare rumen-protected lysine. Rumen-protected methionine is prepared using the same coating process, with the coating material amounting to 18% of the weight of the methionine.

[0079] Step 2, preparation of probiotic preparation: Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium supplemented with 0.15wt% cysteine ​​hydrochloride, modified brain heart infusion medium supplemented with 0.25wt% ox bile salts, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 180r / min for 20h. The culture solution was centrifuged at 5500r / min for 12min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 12% of the cell weight (the protective agent consists of trehalose and skim milk powder in a weight ratio of 1.2:1), and freeze-dried at -55°C and 1Pa under vacuum conditions for 30h;

[0080] Step 3, microencapsulation treatment: the phytosterols are mixed with a wall material composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5 in a weight ratio of 1:3.2 to prepare an aqueous solution with a solid content of 16%, and the mixture is stirred at 52°C and a stirring speed of 250 r / min for 35 minutes. The microcapsules are prepared by spray drying. During the spray drying process, the inlet air temperature is controlled at 185°C, the outlet air temperature is controlled at 82°C, the atomizer speed is 20000±2000 r / min, and the inlet air humidity is 15%. The average particle size of the microcapsules is controlled at 10-15 μm. Scanning electron microscopy shows that the surface porosity of the microcapsules does not exceed 5%;

[0081] Step 4, gradient mixing process: the rumen-protected amino acid complex, complex enzyme preparation, and buffer system are added to the biaxial paddle mixer in the first mixing step and mixed at a speed of 120 r / min for 12 minutes; the probiotic preparation and phytosterol microcapsules are added in the second mixing step, the planetary mixing mode is switched to control the revolution speed to 80 r / min and the rotation speed to 200 r / min, and the mixing is carried out for 22 minutes; the trace element complex is prepared into an aqueous solution with a concentration of 0.6 g / mL in the third mixing step, and the solution is evenly sprayed into the mixer through an atomizing nozzle at a pressure of 0.4 MPa, and the mixing is continued for 10 minutes;

[0082] Step 5, temperature-controlled granulation: the mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 62 ° C, the screw speed is 180 r / min, the granulation pressure is 0.35 MPa, and the obtained granules are sieved through an 85-mesh sieve. The sieve material is crushed and back-mixed with the new mixture at a ratio of 1:3 and then granulated. The granules are subjected to a fluidized bed drying process with an inlet air temperature of 70 ° C and a material fluidization velocity of 0.8 m / s. The moisture content of the granules after drying is not more than 7.5% by Karl Fischer titration, and the surface is coated with 0.2 wt% magnesium stearate.

[0083] This embodiment can gently improve the rumen microecological environment through low-intensity nutritional regulation, reduce the stress response of young animals, and enhance the adaptability of young animals to feed.

[0084] Example 2:

[0085] This embodiment provides a medium-concentration formula suitable for mid-growth ruminants, which balances nutritional regulation effects and cost input by optimizing the coating process and probiotic combination.

[0086] A feed additive for fattening and promoting growth of ruminants, comprising the following components and their mass percentages:

[0087] 25 parts of phytosterols, 45 parts of rumen-protected amino acid complex, 12.5 parts of compound probiotic preparation, 6.75 parts of compound enzyme preparation, 10 parts of trace element complex, 4.4 parts of buffer system; wherein:

[0088] The phytosterols contain 68% beta-sitosterol and 17.5% stigmasterol, which are mixed in a weight ratio of 4:1.

[0089] The rumen-protected amino acid complex consists of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.5:1. The coating material for the rumen-protected lysine is a complex of modified hydrogenated palm oil with a hydroxyl content of 17.5 mol% and lecithin, with the coating material amounting to 23% of the weight of the lysine. The coating material for the rumen-protected methionine is 20% of the weight of the methionine.

[0090] The composite probiotic preparation consists of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans, with a live bacterial count ratio of 3.5:2.5:1:1;

[0091] The complex enzyme preparation contains 40,000 U / g cellulase, 25,000 U / g xylanase, and 12,500 U / g amylase, with the ratio of the three enzyme activities being 3.5:2.5:1.

[0092] The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, with the weight ratio of each component being 5.5:1.5:3.5:1.5:0.5;

[0093] The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, with a weight ratio of 2.5:1.5:0.7.

[0094] A method for preparing a feed additive for fattening and promoting growth of ruminant animals comprises the following steps:

[0095] Step 1: Preparing a protective coating: Modified hydrogenated palm oil with a hydroxyl content of 17.5 mol% and lecithin are mixed in a weight ratio of 5:1, heated to 70°C to melt, lysine raw material is added so that the coating material accounts for 23% of the weight of the lysine, and mixing is carried out at a stirring speed of 250 r / min for 20 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 130 MPa to prepare rumen-protected lysine. Rumen-protected methionine is prepared using the same coating process, with the coating material amounting to 20% of the weight of the methionine.

[0096] Step 2, preparation of probiotic preparation: Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium supplemented with 0.2wt% cysteine ​​hydrochloride, modified brain heart infusion medium supplemented with 0.3wt% ox bile salts, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 200r / min for 21h. The culture solution was centrifuged at 6000r / min for 15min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 13% of the cell weight (the protective agent consists of trehalose and skim milk powder in a weight ratio of 1.2:1), and freeze-dried at -50°C and 2Pa under vacuum conditions for 32h;

[0097] Step 3, microencapsulation treatment: the phytosterols are mixed with a wall material composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5 in a weight ratio of 1:3.5 to prepare an aqueous solution with a solid content of 17.5%, and the mixture is stirred at 55°C and a stirring speed of 300r / min for 40 minutes. The microcapsules are prepared by spray drying. During the spray drying process, the inlet air temperature is controlled at 190°C, the outlet air temperature is controlled at 85°C, the atomizer speed is 20000±2000r / min, and the inlet air humidity is 18%. The average particle size of the microcapsules is controlled at 10-15μm. The moisture absorption rate of the microcapsules does not exceed 8% when placed in an environment with a relative humidity of 75% for 24 hours as tested by a DVS dynamic water vapor sorption instrument;

[0098] Step 4, gradient mixing process: in the first mixing, the rumen-protected amino acid complex, the complex enzyme preparation, and the buffer system are added to the biaxial paddle mixer and mixed at a speed of 130 r / min for 15 minutes; in the second mixing, the probiotic preparation and the phytosterol microcapsules are added, the planetary mixing mode is switched to control the revolution speed to 90 r / min and the rotation speed to 220 r / min, and the mixture is mixed for 25 minutes; in the third mixing, the trace element complex is prepared into an aqueous solution with a concentration of 0.75 g / mL, and the solution is evenly sprayed into the mixer through an atomizing nozzle at a pressure of 0.5 MPa, and the mixing is continued for 12 minutes;

[0099] Step 5, temperature-controlled granulation: the mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 65 ° C, the screw speed is 200 r / min, the granulation pressure is 0.4 MPa, and the obtained granules are sieved with a 90 mesh sieve. The sieve material is crushed and back-mixed with the new mixture at a ratio of 1:3 and then granulated. The granules are subjected to a fluidized bed drying process with an inlet air temperature of 73 ° C and a material fluidization speed of 1.0 m / s. The moisture content of the granules after drying is not more than 7.5% by Karl Fischer titration, and the surface is coated with 0.25 wt% of magnesium stearate.

[0100] This embodiment significantly improves the growth performance of ruminants, promotes the proliferation of beneficial rumen bacteria, and optimizes the efficiency of nutrient absorption by optimizing the functional ingredient ratio and preparation process.

[0101] Example 3:

[0102] This embodiment provides a high-concentration fortified formula suitable for ruminants in the late fattening stage or in scenarios requiring high-intensity fattening, which achieves the goal of rapid fattening by maximizing the concentration of functional ingredients and optimizing process parameters.

[0103] A feed additive for fattening and promoting growth of ruminants, comprising the following components and their mass percentages:

[0104] 28 parts of plant sterols, 48 ​​parts of rumen-protected amino acid complex, 14 parts of compound probiotic preparation, 7.5 parts of compound enzyme preparation, 11 parts of trace element complex, 4.8 parts of buffer system; wherein:

[0105] The phytosterols contain 70% beta-sitosterol and 20% stigmasterol, which are mixed at a weight ratio of 4.5:1.

[0106] The rumen-protected amino acid complex consists of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.8:1. The coating material for the rumen-protected lysine is a complex of modified hydrogenated palm oil with a hydroxyl content of 20 mol% and lecithin, with the coating material amounting to 24% of the weight of the lysine. The coating material for the rumen-protected methionine is 22% of the weight of the methionine.

[0107] The composite probiotic preparation consists of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans, with a live bacterial count ratio of 3.8:2.9:1:1.2;

[0108] The complex enzyme preparation contains 45,000 U / g cellulase, 30,000 U / g xylanase, and 15,000 U / g amylase, with the ratio of the three enzyme activities being 3.7:2.8:1.

[0109] The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, with the weight ratio of each component being 5.8:1.9:3.9:1.9:0.7;

[0110] The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, with a weight ratio of 2.9:1.8:0.9.

[0111] A method for preparing a feed additive for fattening and promoting growth of ruminant animals comprises the following steps:

[0112] Step 1, preparing a protective coating: modified hydrogenated palm oil with a hydroxyl content of 20 mol% and lecithin are mixed in a weight ratio of 5.5:1, heated to 75°C for melting, lysine raw material is added so that the coating material accounts for 24% of the weight of the lysine, and mixing is carried out at a stirring speed of 300 r / min for 25 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 140 MPa to prepare rumen-protected lysine; rumen-protected methionine is prepared using the same coating process, with the coating material amounting to 22% of the weight of the methionine.

[0113] Step 2. Preparation of probiotic preparation: Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium supplemented with 0.25wt% cysteine ​​hydrochloride, modified brain heart infusion medium supplemented with 0.35wt% ox bile salts, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 220r / min for 22h. The culture solution was centrifuged at 6500r / min for 18min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 14% of the cell weight (the protective agent consists of trehalose and skim milk powder in a weight ratio of 1.2:1), and freeze-dried at -45°C and 3Pa under vacuum conditions for 34h.

[0114] Step 3, microencapsulation treatment: the plant sterols were mixed with a wall material composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5 in a weight ratio of 1:3.8 to prepare an aqueous solution with a solid content of 19%, stirred at 58 ° C and a stirring speed of 350 r / min for 45 minutes, and spray-dried to obtain microcapsules. During the spray drying process, the inlet air temperature was controlled at 195 ° C, the outlet air temperature was controlled at 88 ° C, the atomizer speed was 20000 ± 2000 r / min, and the inlet air humidity was 20%, so that the average particle size of the microcapsules was controlled at 10-15 μm. Scanning electron microscopy showed that the surface porosity of the microcapsules did not exceed 5%, and the specific surface area was not less than 50 m 2 / g.

[0115] Step 4, gradient mixing process: add the rumen-protected amino acid complex, complex enzyme preparation, and buffer system to the double-shaft paddle mixer in the first mixing, and mix at a speed of 140r / min for 18 minutes; add the probiotic preparation and plant sterol microcapsules in the second mixing, switch to the planetary mixing mode, control the revolution speed to 100r / min and the rotation speed to 240r / min, and mix for 28 minutes; the trace element complex is prepared into an aqueous solution with a concentration of 0.9g / mL, and is evenly sprayed into the mixer through an atomizing nozzle at a pressure of 0.6MPa, and the mixing is continued for 15 minutes.

[0116] Step 5, temperature-controlled granulation: the mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 68 ° C, the screw speed is 220 r / min, the granulation pressure is 0.45 MPa, and the obtained granules are sieved with a 95 mesh sieve. The sieve material is crushed and back-mixed with the new mixture at a ratio of 1:3 and then granulated. The granules are subjected to a fluidized bed drying process with an inlet air temperature of 75 ° C and a material fluidization speed of 1.2 m / s. The moisture content of the granules after drying is not more than 7.5% by Karl Fischer titration, and the surface is coated with 0.3 wt% of magnesium stearate.

[0117] This embodiment significantly improves the fattening efficiency of ruminants, enhances the stability and enzyme activity of probiotics, optimizes the rumen microecological environment, shortens the fattening cycle and improves the health level of the body through high-concentration functional ingredients and refined processes.

[0118] Example 4:

[0119] This example verifies the effects of ruminant fattening and growth-promoting feed additives with different component ratios on the growth performance and rumen microecology of weaned lambs, providing data support for formula optimization and application.

[0120] Experimental animals: 60 healthy weaned Hu sheep lambs of similar weight (25±2kg) were randomly divided into 4 groups, with 15 lambs in each group.

[0121] Group processing:

[0122] Control group: fed with basic diet without adding this feed additive;

[0123] Experimental group 1: fed the basic diet + Example 1, 22 parts of plant sterols, 42 parts of rumen-protected amino acid complex, 11 parts of compound probiotic preparation, 6 parts of compound enzyme preparation, 9 parts of trace element complex, and 4 parts of buffer system;

[0124] Experimental group 2: fed the basic diet + 25 parts of the plant sterols of Example 2, 45 parts of the rumen-protected amino acid complex, 12.5 parts of the compound probiotic preparation, 6.75 parts of the compound enzyme preparation, 10 parts of the trace element complex, and 4.4 parts of the buffer system;

[0125] Experimental group 3: fed with the basic diet + 28 parts of the plant sterols of the third component of Example, 48 parts of the rumen-protected amino acid complex, 14 parts of the compound probiotic preparation, 7.5 parts of the compound enzyme preparation, 11 parts of the trace element complex, and 4.8 parts of the buffer system.

[0126] The amount of additives added was 1.5% of the basic diet weight, and the experimental period was 60 days.

[0127] During the experiment, the animals were fed three times a day (06:00, 12:00, and 18:00), with free access to food and water. The pen temperature was maintained at 18-22°C, the humidity at 55%-65%, and disinfection was carried out regularly.

[0128] Growth performance:

[0129] Fasting body weight was measured weekly to calculate daily weight gain (ADG); daily feed intake was recorded to calculate feed conversion ratio (FCR = feed intake / weight gain).

[0130] Rumen microecology:

[0131] On the 30th and 60th days of the experiment, rumen fluid was collected through rumen fistula, and the diversity of rumen microbial flora was detected by high-throughput sequencing; the pH value and volatile fatty acid (VFA) content of rumen fluid were detected (high-performance liquid chromatography).

[0132] Probiotic colonization ability: After the experiment, lamb feces were collected and the number of probiotics (Lactobacillus plantarum and Enterococcus faecalis) was detected by MPN-PCR method.

[0133] Table 1 Experimental data of control group

[0134]

[0135]

[0136] Experimental data show that the daily weight gain of weaned lambs in the experimental group supplemented with this ruminant animal fattening and growth-promoting feed additive was significantly improved compared with the control group. The low, medium and high concentration groups increased by 11.9%, 23.8% and 35.7% respectively. The feed conversion rate was optimized, and the high concentration group dropped to 3.3±0.1. The rumen pH value approached the healthy range, and the acetic acid / propionic acid ratio gradually decreased from 3.8:1 in the control group to 2.9:1. The number of fecal probiotics increased significantly with the increase of additive concentration. The number of plant lactobacillus in the high concentration group was 5 times that of the control group, indicating that the additive can effectively improve the growth performance of ruminants and improve the rumen microecology, and the high concentration formula has better fattening effect.

[0137] In summary, the preparation method of the feed additive for fattening and promoting growth of ruminant animals adopts a complex of modified hydrogenated palm oil and lecithin as a rumen-protected amino acid coating material, uses a high-pressure homogenizer to perform multi-stage homogenization treatment, so as to make the coating layer structure denser, enhance the decomposition ability of anti-rumen microbial lipase through the synergistic effect of the composite material, optimizes the particle distribution in combination with the refined homogenization process, improves the release controllability and absorption efficiency of amino acids in the small intestine, and solves the problems of easy decomposition and extensive particle size control of traditional single coating materials.

[0138] In addition, the preparation method of the feed additive for fattening and promoting growth of ruminant animals comprises the following steps: forming a composite probiotic preparation of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum and Bacillus coagulans, adding cysteine ​​hydrochloride and ox bile salt to the improved culture medium to optimize the bacterial culture conditions, combining trehalose and skim milk powder composite protective agent and low-temperature vacuum freeze-drying process, improving the diversity of the bacterial community through the synergistic effect of multiple bacterial strains, enhancing the stress resistance of the bacterial body by using culture medium components, maintaining the activity of live bacteria with the help of protective agent and freeze-drying process, solving the problems of single probiotic strain, insufficient high temperature resistance and rumen colonization ability, and solving the problems of easy decomposition of rumen-protected amino acids in existing additives, single probiotic strain and insufficient colonization ability.

[0139] The relevant modules involved in this system are all hardware system modules or functional modules that combine computer software programs or protocols with hardware in the existing technology. The computer software programs or protocols involved in the functional modules are themselves technologies that are well known to those skilled in the art and are not improvements to this system. The improvements to this system are the interaction or connection relationships between the modules, that is, improvements to the overall structure of the system to solve the corresponding technical problems to be solved by this system.

[0140] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A feed additive for fattening and promoting growth of ruminant animals, characterized in that: The invention comprises the following components and their mass percentages: 22-28 parts of plant sterols, 42-48 parts of rumen-protected amino acid complex, 11-14 parts of compound probiotic preparation, 6-7.5 parts of compound enzyme preparation, 9-11 parts of trace element complex and 4-4.8 parts of buffer system; The β-sitosterol content in the phytosterol is not less than 65%, and the stigmasterol content is 15-20%; The rumen-protected amino acid complex is composed of rumen-protected lysine and rumen-protected methionine in a weight ratio of 2.2-2.8:1, wherein the coating material of the rumen-protected lysine is a complex of modified hydrogenated palm oil with a hydroxyl content of 15-20 mol% and lecithin, and the amount of the coating material is 22-24% of the weight of the lysine; The composite probiotic preparation is composed of Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum and Bacillus coagulans, and the ratio of viable bacteria count is 3.2-3.8:2.1-2.9:1:0.8-1.2; The complex enzyme preparation comprises cellulase with an enzyme activity of 35,000-45,000 U / g, xylanase with an enzyme activity of 20,000-30,000 U / g, and amylase with an enzyme activity ratio of 10,000-15,000 U / g, with the ratio of the three enzymes being 3.3-3.7:2.2-2.8:1; The trace element complex is composed of ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate and yeast selenium, and the weight ratio of each component is 5.2-5.8:1.1-1.9:3.1-3.9:1.1-1.9:0.3-0.7; The buffer system consists of sodium bicarbonate, sodium dihydrogen phosphate and sodium citrate, and the weight ratio of the three is 2.1-2.9:1.2-1.8:0.5-0.

9.

2. The ruminant fattening and growth-promoting feed additive according to claim 1, characterized in that: The phytosterols are separated and prepared by silica gel column chromatography, using n-hexane-ethyl acetate with a volume ratio of 8:2 as an eluent, collecting the β-sitosterol elution peak with a retention time of 28-32 minutes, and the purity after freeze crystallization is not less than 98%; The stigmasterol is extracted by supercritical CO2 at an extraction pressure of 25-30 MPa and a temperature of 45-50° C., with a yield of not less than 92%. β-sitosterol and stigmasterol are compounded at a weight ratio of 3.5-4.5:

1.

3. The ruminant fattening and growth-promoting feed additive according to claim 1, characterized in that: The cellulase is derived from a mutant strain of Trichoderma reesei QM6a, and is purified by an ammonium sulfate salting-out method after liquid fermentation. The salting-out saturation is 30-50%, the enzyme protein purity is not less than 90%, and the enzyme activity retention rate is not less than 95% at an optimal pH of 5.8-6.

2. The xylanase is introduced with histidine at the 125th amino acid by using site-directed mutagenesis technology, so that its optimum action temperature is increased to 50-55° C., and the residual enzyme activity is not less than 70% after being treated at 60° C. for 30 minutes.

4. A method for preparing a feed additive for fattening and promoting growth of ruminant animals, characterized in that: The method comprises the ruminant fattening and growth-promoting feed additive according to claims 1 to 3, and the following steps: S1. Preparation of protective coating: Modified hydrogenated palm oil having a hydroxyl content of 15-20 mol% and lecithin are mixed in a weight ratio of 4.5-5.5:1, heated to 65-75° C. to melt, lysine raw material is added so that the coating material accounts for 22-24% of the weight of the lysine, and the mixture is stirred at a speed of 200-300 r / min for 15-25 minutes. The mixture is homogenized three times in a high-pressure homogenizer at a pressure of 120-140 MPa to prepare rumen-protected lysine. The same coating process is used to prepare rumen-protected methionine, with the coating material dosage being 18-22% of the weight of methionine; S2. Preparation of probiotic preparations: Lactobacillus plantarum, Enterococcus faecalis, Clostridium butyricum, and Bacillus coagulans were inoculated into modified MRS medium, modified brain heart infusion medium, modified cooked meat medium, and LB medium, respectively, and cultured at 37±0.5°C and 180-220 rpm for 20-22 h. The culture broth was centrifuged at 5500-6500 rpm for 12-18 min to collect the cells, washed twice with physiological saline containing 5% trehalose, and mixed with a protective agent accounting for 12-14% of the cell weight, wherein the protective agent consisted of trehalose and skim milk powder in a weight ratio of 1.2:1, and freeze-dried at -55-45°C and a vacuum condition of 1-3 Pa for 30-34 h. S3. Microencapsulation treatment: The phytosterols are mixed with a wall material, wherein the wall material is composed of gum arabic, gelatin, and β-cyclodextrin in a weight ratio of 1:1:0.5, and the weight ratio of the phytosterols to the wall material is 1:3.2-3.8, to prepare an aqueous solution with a solid content of 16-19%, stirring at 52-58° C. and a stirring speed of 250-350 r / min for 35-45 minutes, and spray drying to obtain microcapsules. During the spray drying process, the air inlet temperature is controlled at 185-195° C., and the air outlet temperature is controlled at 82-88° C. S4, gradient mixing process: Primary mixing: Add the rumen-protected amino acid complex, complex enzyme preparation, and buffer system into a twin-shaft paddle mixer and mix at a speed of 120-140 r / min for 12-18 minutes; Secondary mixing: Add probiotic preparation and phytosterol microcapsules, switch to planetary mixing mode, control the revolution speed to 80-100 r / min and the rotation speed to 200-240 r / min, and mix for 22-28 minutes; Three-stage mixing: prepare the trace element complex into an aqueous solution with a concentration of 0.6-0.9 g / mL, spray it evenly into the mixer through an atomizing nozzle at a pressure of 0.4-0.6 MPa, and continue mixing for 10-15 minutes; S5. Temperature controlled granulation: The mixture is processed by a twin-screw granulator, the granulation temperature is controlled at 62-68°C, the screw speed is 180-220r / min, the granulation pressure is 0.35-0.45MPa, and the obtained granules are sieved through an 85-95 mesh sieve. The sieve material is crushed and then mixed with the new mixture in a ratio of 1:3 and granulated again.

5. The method for preparing a ruminant fattening and growth-promoting feed additive according to claim 4, characterized in that: The modified MRS medium is added with 0.15-0.25 wt% cysteine ​​hydrochloride to maintain the integrity of Lactobacillus plantarum cells through sulfhydryl groups, thereby increasing the survival rate of bacteria in the logarithmic growth phase by 20%; The modified brain heart infusion medium is added with 0.25-0.35 wt% of ox bile salts to induce Enterococcus faecalis to produce bile salt hydrolase, thereby enhancing its colonization ability in the high bile salt environment of the rumen and increasing the bile salt tolerance of Enterococcus faecalis to 0.5 wt%.

6. The method for preparing a ruminant fattening and growth-promoting feed additive according to claim 4, characterized in that: The spray drying process uses a pressure atomizer with a speed of 20000±2000r / min and an inlet air humidity of 15-20%, so that the average particle size of the microcapsules is controlled at 10-15μm. Scanning electron microscopy shows that the surface porosity of the microcapsules does not exceed 5% and the specific surface area is not less than 50m 2 / g; The contact time between the atomized droplets and the hot air is controlled at 0.8-1.2 seconds. The DVS dynamic water vapor adsorption instrument tests that the moisture absorption rate of the microcapsules does not exceed 8% when placed in an environment with a relative humidity of 75% for 24 hours.

7. The method for preparing a ruminant fattening and growth-promoting feed additive according to claim 4, characterized in that: The high-pressure homogenizer adopts a two-stage homogenizing valve structure, the cone angle of the first-stage homogenizing valve is 60°±5°, and the cone angle of the second-stage homogenizing valve is 45°±5°. The D90 of the coated particles after homogenization is monitored by a laser particle size analyzer. ≤12 μm, and the particle size distribution span is (D90-D10) / D50≤1.5; During the homogenization process, the temperature of the feed liquid is controlled at 40-45°C by jacket circulating water to avoid thermal degradation of the lysine amino group. HPLC testing shows that the amino acid purity is not less than 99.2%.

8. The method for preparing a ruminant fattening and growth-promoting feed additive according to claim 4, characterized in that: The particles are dried using a fluidized bed drying process with an air inlet temperature of 70-75°C and a material fluidization velocity of 0.8-1.2 m / s. The moisture content of the particles after drying is no more than 7.5% as determined by Karl Fischer titration. The surface is coated with 0.2-0.3 wt% of magnesium stearate, which improves moisture resistance by 30%. The number of probiotics is detected by MPN-PCR method, wherein the spore rate of Bacillus coagulans is not less than 90%, and the viable bacteria survival rate after heat shock treatment at 65° C. for 10 minutes is not less than 85%, which meets the detection requirements of AOAC official analysis method 2015.12.

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