Preparation method and application of fat powder capable of promoting absorption of young animals

Abstract

The invention relates to the technical field of animal feed, and discloses a preparation method and application of fat powder for promoting absorption of young animals. According to the fat powder, 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride (OPL), soybean oil and egg yolk lecithin are adopted as an inner core fat library, ethyl cellulose-stearic acid is adopted as a middle wrapping layer, and linolenic acid and sodium butyrate are adopted as shell components. By preferentially releasing sodium butyrate, expression of lipase is promoted, fat is slowly released, and the problem of low fat absorption rate caused by insufficient lipase secretion is prevented. The problem of grease digestion disorder caused by insufficient lipase secretion in the early weaning stage of young animals is solved. Fat powder and young animal basal feed are matched according to a certain proportion to obtain the feed for promoting the young animals to absorb fat, and a method is provided for feed production of the young animals.

Description

Technical Field

[0001] This invention belongs to the field of animal feed technology, and particularly relates to a method for preparing and applying fat powder that promotes absorption by young animals. Background Technology

[0002] In a broad sense, fats, also known as lipids, include true lipids (triglycerides) and phospholipids (phospholipids, sterols, waxes). These compounds are important components of livestock and poultry tissues and have multiple nutritional functions. Fat is the best form of energy storage in livestock and poultry, meeting their requirements for high energy concentrations, improving amino acid digestibility, and inhibiting the conversion of glucose and other precursors into fat. It provides essential fatty acids (linoleic acid, linolenic acid, arachidonic acid, etc.) and acts as a solvent for fat-soluble vitamins and certain hormones, promoting their absorption and utilization. In special environments such as high temperatures, it can reduce stress responses in livestock and poultry. Vegetable oils have a laxative effect, with castor oil having the strongest laxative effect, followed by cottonseed oil and soybean oil. Soybean oil and peanut oil can also treat intestinal obstruction caused by ascariasis.

[0003] In livestock and poultry production, besides serving as an energy source to meet animal needs, oils and fats also have several important physiological functions. ① As a solvent for fat-soluble nutrients, they facilitate the digestion and absorption of these nutrients. ② An important source of metabolic water in the body; the oxidation of fat in animals provides both energy and water. Each gram of fat oxidation produces 67%–83% more water than carbohydrates and about 1.5 times more than protein. ③ Emulsifying phospholipids; because phospholipid molecules contain both hydrophilic phosphate groups and hydrophobic fatty acid chains, they possess emulsifying properties. They promote the formation of a suitable oil-water emulsion environment in the digestive tract and play an important role in the transport of lipids in the blood and the transmembrane transport of nutrients. ④ As a source of essential fatty acids in animals, linoleic acid and linolenic acid in oils and fats cannot be synthesized by the animal body and are indispensable for cell structure and metabolism, and must be obtained from feed.

[0004] Early-weaned piglets have underdeveloped digestive and immune systems, resulting in insufficient secretion of enzymes and gastric acid in the digestive tract. This often leads to what is known as "early weaning syndrome," characterized by decreased appetite, indigestion, low feed utilization, poor disease resistance, diarrhea, poor neurological function, and slow growth. Many factors contribute to post-weaning growth retardation in piglets, with decreased enzyme activity in the digestive system being a significant one. During the first four weeks of life, the activity of digestive enzymes (amylase, protease, and lipase, etc.) in the stomach, pancreas, and small intestine increases dramatically. After weaning, the activity of various pancreatic enzymes decreases to about one-third of pre-weaning levels, and the activity of enzymes in the small intestine also declines, only returning to pre-weaning levels about two weeks later. Therefore, regulating the activity of gastrointestinal digestive enzymes in weaned piglets through nutrition is crucial for promoting nutrient digestion and absorption.

[0005] Fat is a high-energy substance and an ideal energy supplement. The digestion and absorption of fat is a complex process influenced by various factors, including feed, the animal's physiological condition, and the environment. When designing diet formulations, these influencing factors should be fully considered to maximize fat utilization and improve piglet production performance. Currently, the application of fat in weaned piglet diets is generally accepted by most scholars, with piglets showing poor fat utilization in the first 1-2 weeks after weaning, followed by a significant improvement after 2 weeks. Whether the initial addition of fat at weaning induces lipase development remains controversial. How to utilize high-energy substances to improve piglet production performance and fully leverage the physiological functions of fat is key to fat utilization and warrants further in-depth research. Currently, the main approach to improving fat absorption in young animals is by adding lipase to piglet and broiler feed. However, excessive addition of exogenous lipase may inhibit the expression of endogenous lipase, leading to lipase dependence and requiring continuous long-term supplementation. Furthermore, lipase is expensive and easily destroyed by stomach acid during administration, further increasing feed costs due to the need for lipase protection. Summary of the Invention

[0006] This invention aims to provide a method for preparing and applying a fat powder that promotes absorption in young animals. The fat powder uses 1-oleic-2-palmitic-3-linoleic acid triglycerides, soybean oil, and egg yolk lecithin as the core fat reservoir, employs ethyl cellulose-stearic acid as the intermediate coating layer, and uses linolenic acid and sodium butyrate as the outer shell components for preferential release. By mixing the fat powder with the basic feed for young animals in a certain proportion, it can promote the expression of intestinal lipases in young animals, increasing their absorption and metabolism of fat. This allows the fat to be gradually degraded in the upper small intestine, reserving time for enzyme expression and preventing premature exposure of the fat.

[0007] To achieve the above objectives, the present invention provides a method for preparing fat powder that promotes absorption in young animals, comprising the following steps:

[0008] S1. Place 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride, soybean oil and egg yolk lecithin into a three-necked flask, purge with nitrogen, and stir in a water bath until a homogeneous oil phase is formed; in another beaker, add hydrogenated soybean lecithin and cholesterol, pour in chloroform, and stir until completely dissolved to obtain the membrane solution;

[0009] S2. Pour the homogeneous oil phase into the membrane material solution and stir to form a mixture; transfer the mixture to a flask of a rotary evaporator and rotary evaporate until the chloroform is completely evaporated, forming a homogeneous lipid film on the inner wall of the flask;

[0010] S3. Add PBS buffer to the flask in S2, and stir in a water bath for the second time until the lipid film is completely hydrated to form a crude liposome suspension; transfer the crude liposome suspension to an ultrasonic disruptor and sonicate to obtain nano-sized liposomes; spray dry the nano-sized liposomes to obtain liposome powder.

[0011] S4. Ethyl cellulose and stearic acid are dissolved in anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome micropowder is added to a fluidized bed, sprayed with the ethyl cellulose-stearic acid solution, and dried for the first time to obtain bilayer microspheres.

[0012] S5. Sodium butyrate, linolenic acid, and Lactobacillus reuteri were dispersed in deionized water, and gelatin and low-acyl gellan gum were added. The mixture was stirred and dissolved to obtain a shell gel. The bilayer microspheres were added to a fluidized bed, the shell gel was sprayed on, and the mixture was dried a second time to obtain a fat powder that promotes absorption in young animals.

[0013] Preferably, in step S1, the mass ratio of 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride, soybean oil, and egg yolk lecithin is 50:(25-35):(5-15); the water bath stirring temperature is 45-55°C, and the stirring speed for the first water bath stirring is 250-350 rpm; the mass ratio of hydrogenated soybean lecithin, cholesterol, and 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride is (5-7):(1-3):50; the ratio of chloroform to 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride is (3.5-4.5) mL:1 g; and the stirring speed until complete dissolution is 150-250 rpm.

[0014] Preferably, in step S2, the ratio of the homogeneous oil phase to the film material solution is 90g:(175~225)mL; the stirring speed for forming the mixture is 250~350rpm, and the time is 15~25min; the rotary evaporation temperature is 30~40℃, the vacuum degree is 0.07~0.09MPa, and the rotation speed is 50~70rpm.

[0015] Preferably, in step S3, the molar concentration of the PBS buffer is 0.01–0.03 M, the pH is 7.2–7.6, and the ratio of the PBS buffer to the homogeneous oil phase in step S2 is (900–1000) mL:90 g; the second water bath stirring temperature is 35–40 °C, and the rotation speed is 150–250 rpm; the ultrasonic power is 250–350 W, and the time is 8–12 min; the spray drying inlet air temperature is 55–65 °C, the outlet air temperature is 30–40 °C, and the feed rate is 8–12 mL / min.

[0016] Preferably, in step S4, the ratio of ethyl cellulose, stearic acid, and anhydrous ethanol is 1 g:(0.8-1.2) g:(90-110) mL; the ratio of liposome micropowder to ethyl cellulose-stearic acid solution is 95 g:(450-550) mL; the fluidized bed inlet air temperature is 35-45°C; the spraying rate is 4-6 mL / min; and the first drying temperature is 35-45°C for 1.5-2.5 h.

[0017] Preferably, in step S5, the ratio of sodium butyrate, linolenic acid, Lactobacillus reuteri, and deionized water is (0.4–0.6) g:(1.5–2.5) g:(0.5–1.5) g:500 mL; the ratio of gelatin, low-acyl gellan gum, and deionized water is (15–25) g:(4–6) g:500 mL; the fluidized bed inlet air temperature is 35–45°C; the spraying rate is 6–10 mL / min; and the second drying temperature is 30–40°C, and the time is 1.0–2.0 h.

[0018] The present invention also provides an application of a fat powder that promotes absorption in young animals, wherein the fat powder that promotes absorption in young animals is used as an additive in the feed of young animals.

[0019] Preferably, the feed for young animals is a basic feed powder for young animals, and the mass ratio of the feed additive for young animals to the basic feed powder for young animals is 1:(8-10).

[0020] Compared with the prior art, the beneficial effects of the present invention are reflected in:

[0021] (1) This invention achieves a breakthrough through the precise spatiotemporal regulation of a three-layer core-shell structure: the outer shell is made of gelatin-gellan gel material, which rapidly releases sodium butyrate, linolenic acid and probiotics in the stomach and upper small intestine, thus initiating the gene expression of lipase in advance and regulating the signaling pathways related to fat metabolism; the middle layer is made of ethyl cellulose-stearic acid coating layer, which gradually degrades in the upper small intestine, delaying the release of fat and reserving time for enzyme expression; the liposome membrane of the core slowly releases oil in the small intestine, matching the peak expression of lipase; thus solving the problem of fat digestion disorder caused by insufficient enzymes in the early stage of weaning in young animals from a mechanistic perspective.

[0022] (2) In this invention, 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride (OPL), soybean oil, and egg yolk lecithin are used as the main components of fat in fat powder. OPL structured lipid is a structured lipid designed by mimicking the natural distribution of triglycerides in breast milk. It can improve gastric emptying rate and intestinal absorption efficiency, significantly increase the abundance of probiotics such as Bifidobacterium, and inhibit the growth of pathogenic bacteria such as Escherichia coli. It helps young animals absorb fat. The emulsifying effect of egg yolk lecithin reduces the dependence of fat on bile acids. Attached Figure Description

[0023] Figure 1 This is a photograph of the fat powder used in Example 1 to promote absorption in young animals.

[0024] Figure 2 This is a photograph of the feed containing fat powder that promotes absorption in young animals, as shown in Example 4.

[0025] Figure 3 A statistical graph showing the relative expression levels of AMPK in jejunal tissue detected by RT-PCR;

[0026] Figure 4 A statistical graph showing the relative expression levels of PPARα in jejunal tissue detected by RT-PCR;

[0027] Figure 5 A statistical graph showing the activity of porcine pancreatic lipase in intestinal contents detected by ELISA. Detailed Implementation

[0028] The following embodiments are only used to illustrate the technical solutions of the present invention more clearly, and should not be used to limit the scope of protection of the present invention.

[0029] The main compounds used in the examples and comparative examples were all commercially available products and were not subjected to any further purification treatment.

[0030] Example 1

[0031] A method for preparing a fat powder that promotes absorption in young animals includes the following steps:

[0032] S1. Take 50g OPL, 30g soybean oil and 10g egg yolk lecithin into a three-necked flask, purge with nitrogen, place in a 50℃ water bath, and stir at 300rpm until a homogeneous oil phase is formed; in another beaker, add 6g hydrogenated soybean lecithin and 2g cholesterol, pour in 200mL chloroform, and stir at 200rpm until completely dissolved to obtain the membrane solution.

[0033] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 300 rpm for 20 min to form a mixture; transfer the mixture to a flask of a rotary evaporator and evaporate it at 35℃ and 0.08 MPa at 60 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0034] S3. Add 1000 mL of 0.02 M PBS buffer (pH 7.4) to the flask from S2. Incubate at 37°C and stir at 200 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic disruptor and sonicate at 300 W for 10 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 60°C, an outlet air temperature of 35°C, and a feed rate of 10 mL / min to obtain liposome micropowder.

[0035] S4.5g of ethyl cellulose and 5g of stearic acid were dissolved in 500mL of anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome micropowder was added to a fluidized bed with an inlet air temperature of 40℃. The ethyl cellulose-stearic acid solution was sprayed at 5mL / min and dried at 40℃ for 2h to obtain bilayer microspheres.

[0036] S5.0.5g sodium butyrate, 2g linolenic acid, and 1g Lactobacillus reuteri were dispersed in 500mL deionized water; 20g gelatin and 5g low-acyl gellan gum were added and stirred to dissolve, resulting in a shell gel; bilayer microspheres were added to a fluidized bed with an inlet air temperature of 40℃, and the shell gel was sprayed at 8mL / min. The mixture was then dried at 35℃ for 1.5h to obtain a fat powder that promotes fat absorption efficiency in young animals (e.g., ...). Figure 1 (As shown).

[0037] Example 2

[0038] A method for preparing a fat powder that promotes absorption in young animals includes the following steps:

[0039] S1. Take 50g OPL, 25g soybean oil and 15g egg yolk lecithin into a three-necked flask, purge with nitrogen, and stir at 350rpm in a 45℃ water bath until a homogeneous oil phase is formed; take another beaker, add 5g hydrogenated soybean lecithin and 3g cholesterol, pour in 175mL chloroform, and stir at 150rpm until completely dissolved to obtain the membrane solution.

[0040] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 350 rpm for 15 min to form a mixture; transfer the mixture to a flask in a rotary evaporator and evaporate it at 30℃ and 0.09 MPa at 50 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0041] S3. Add 1100 mL of 0.01 M PBS buffer (pH 7.2) to the flask from S2. Incubate at 35°C and stir at 250 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic disruptor and sonicate at 250 W for 12 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 55°C, an outlet air temperature of 30°C, and a feed rate of 8 mL / min to obtain liposome micropowder.

[0042] S4.5g ethyl cellulose and 4g stearic acid were dissolved in 450mL anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome powder was added to a fluidized bed with an inlet air temperature of 35℃. The ethyl cellulose-stearic acid solution was sprayed at a rate of 4mL / min and dried at 35℃ for 2.5h to obtain bilayer microspheres.

[0043] S5.0.4g sodium butyrate, 1.5g linolenic acid and 1.5g Lactobacillus reuteri were dispersed in 500mL deionized water; 25g gelatin and 4g low-acyl gellan gum were added and stirred to dissolve to obtain the outer shell gel; the bilayer microspheres were added to a fluidized bed with an inlet air temperature of 35℃, and the outer shell gel was sprayed at 6mL / min. The mixture was then dried at 30℃ for 2h to obtain fat powder that promotes absorption in young animals.

[0044] Example 3

[0045] A method for preparing a fat powder that promotes absorption in young animals includes the following steps:

[0046] S1. Take 50g OPL, 35g soybean oil and 5g egg yolk lecithin into a three-necked flask, purge with nitrogen, place in a 55℃ water bath, and stir at 250rpm until a homogeneous oil phase is formed; in another beaker, add 7g hydrogenated soybean lecithin and 1g cholesterol, pour in 225mL chloroform, and stir at 250rpm until completely dissolved to obtain the membrane solution.

[0047] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 250 rpm for 25 min to form a mixture; transfer the mixture to a flask in a rotary evaporator and evaporate it at 40℃ and 0.07 MPa at 70 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0048] S3. Add 900 mL of 0.03 M PBS buffer (pH 7.6) to the flask from S2. Incubate at 40°C and stir at 150 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic homogenizer and sonicate at 350 W for 8 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 65°C, an outlet air temperature of 40°C, and a feed rate of 12 mL / min to obtain liposome micropowder.

[0049] S4.5g ethyl cellulose and 6g stearic acid were dissolved in 550mL anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome powder was added to a fluidized bed with an inlet air temperature of 45℃. The ethyl cellulose-stearic acid solution was sprayed at a rate of 6mL / min and dried at 45℃ for 1.5h to obtain bilayer microspheres.

[0050] S5.0.6g sodium butyrate, 2.5g linolenic acid and 0.5g Lactobacillus reuteri were dispersed in 500mL deionized water; 15g gelatin and 6g low-acyl gellan gum were added and stirred to dissolve to obtain the outer shell gel; the bilayer microspheres were added to a fluidized bed with an inlet air temperature of 45℃, and the outer shell gel was sprayed at 10mL / min. The mixture was then dried at 40℃ for 1.0h to obtain fat powder that promotes absorption in young animals.

[0051] Example 4

[0052] A method for preparing feed containing fat powder that promotes absorption in young animals includes the following steps:

[0053] 580g corn flour, 200g soybean meal, 80g wheat bran, 30g fish meal, 10g bone meal, and 5g salt were mixed to obtain a basic pig feed powder. 100g of the fat powder prepared in Example 1 was mixed with 900g of the basic pig feed powder, and pelleted using a pelleting machine to obtain a feed containing fat powder that promotes absorption in young animals (e.g., ...). Figure 2 (As shown).

[0054] Example 5

[0055] A method for preparing feed containing fat powder that promotes absorption in young animals includes the following steps:

[0056] 580g of corn flour, 200g of soybean meal, 80g of wheat bran, 30g of fish meal, 10g of bone meal, and 5g of salt were mixed to obtain a basic pig feed powder. 100g of fat powder prepared in Example 2 was mixed with 900g of basic pig feed powder and pelleted by a pelleting machine to obtain a feed containing fat powder that promotes absorption by young animals.

[0057] Example 6

[0058] A method for preparing feed containing fat powder that promotes absorption in young animals includes the following steps:

[0059] 580g of corn flour, 200g of soybean meal, 80g of wheat bran, 30g of fish meal, 10g of bone meal, and 5g of salt were mixed to obtain a basic pig feed powder. 100g of fat powder prepared in Example 3 was mixed with 900g of basic pig feed powder and pelleted by a pelleting machine to obtain a feed containing fat powder that promotes absorption by young animals.

[0060] Comparative Example 1

[0061] A method for preparing sustained-release fat powder includes the following steps:

[0062] S1. Take 50g OPL, 30g soybean oil and 10g egg yolk lecithin into a three-necked flask, purge with nitrogen, place in a 50℃ water bath, and stir at 300rpm until a homogeneous oil phase is formed; in another beaker, add 6g hydrogenated soybean lecithin and 2g cholesterol, pour in 200mL chloroform, and stir at 200rpm until completely dissolved to obtain the membrane solution.

[0063] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 300 rpm for 20 min to form a mixture; transfer the mixture to a flask of a rotary evaporator and evaporate it at 35℃ and 0.08 MPa at 60 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0064] S3. Add 1000 mL of 0.02 M PBS buffer (pH 7.4) to the flask from S2. Incubate at 37°C and stir at 200 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic disruptor and sonicate at 300 W for 10 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 60°C, an outlet air temperature of 35°C, and a feed rate of 10 mL / min to obtain liposome micropowder.

[0065] S4.5g of ethyl cellulose and 5g of stearic acid were dissolved in 500mL of anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome powder was added to a fluidized bed with an inlet air temperature of 40℃. The ethyl cellulose-stearic acid solution was sprayed at a rate of 5mL / min and dried at 40℃ for 2h to obtain fat powder.

[0066] Comparative Example 2

[0067] A method for preparing liposome fat powder includes the following steps:

[0068] S1. Take 50g OPL, 30g soybean oil and 10g egg yolk lecithin into a three-necked flask, purge with nitrogen, place in a 50℃ water bath, and stir at 300rpm until a homogeneous oil phase is formed; in another beaker, add 6g hydrogenated soybean lecithin and 2g cholesterol, pour in 200mL chloroform, and stir at 200rpm until completely dissolved to obtain the membrane solution.

[0069] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 300 rpm for 20 min to form a mixture; transfer the mixture to a flask of a rotary evaporator and evaporate it at 35℃ and 0.08 MPa at 60 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0070] S3. Add 1000 mL of 0.02 M PBS buffer (pH 7.4) to the flask from S2. Incubate at 37°C and stir at 200 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic disruptor and sonicate at 300 W for 10 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 60°C, an outlet air temperature of 35°C, and a feed rate of 10 mL / min to obtain fat powder.

[0071] Comparative Example 3

[0072] A method for preparing liposome fat powder includes the following steps:

[0073] S1. Take 90g of soybean oil and put it into a three-necked flask. Purge with nitrogen and stir at 300rpm in a 50℃ water bath until a homogeneous oil phase is formed. In another beaker, add 6g of hydrogenated soybean lecithin and 2g of cholesterol, pour in 200mL of chloroform, and stir at 200rpm until completely dissolved to obtain the membrane solution.

[0074] S2. Slowly pour the oil phase from S1 into the membrane material solution and stir at 300 rpm for 20 min to form a mixture; transfer the mixture to a flask of a rotary evaporator and evaporate it at 35℃ and 0.08 MPa at 60 rpm until the chloroform is completely evaporated and a uniform lipid film is formed on the inner wall of the flask.

[0075] S3. Add 1000 mL of 0.02 M PBS buffer (pH 7.4) to the flask from S2. Incubate at 37°C and stir at 200 rpm until the lipid film is completely hydrated, forming a crude liposome suspension. Transfer the suspension to an ultrasonic disruptor and sonicate at 300 W for 10 min to obtain nano-sized liposomes. Spray dry the liposome suspension using a spray dryer with an inlet air temperature of 60°C, an outlet air temperature of 35°C, and a feed rate of 10 mL / min to obtain fat powder.

[0076] Comparative Example 4

[0077] A method for preparing animal feed containing fat powder, comprising the following steps:

[0078] Take 100g of the fat powder prepared in Comparative Example 1, mix it with 900g of basic pig feed powder, and granulate it using a pelleting machine to obtain animal feed containing fat powder.

[0079] Comparative Example 5

[0080] A method for preparing animal feed containing fat powder, comprising the following steps:

[0081] 580g of corn flour, 200g of soybean meal, 80g of wheat bran, 30g of fish meal, 10g of bone meal, and 5g of salt were mixed to obtain a basic pig feed powder. 100g of fat powder prepared in Comparative Example 2 was mixed with 900g of basic pig feed powder and pelleted by a pelleting machine to obtain animal feed containing fat powder.

[0082] Comparative Example 6

[0083] A method for preparing animal feed containing fat powder, comprising the following steps:

[0084] 580g of corn flour, 200g of soybean meal, 80g of wheat bran, 30g of fish meal, 10g of bone meal, and 5g of salt were mixed to obtain a basic pig feed powder. 100g of fat powder prepared in Comparative Example 3 was mixed with 900g of basic pig feed powder and pelleted using a pelleting machine to obtain animal feed containing fat powder.

[0085] Comparative Example 7

[0086] 580g of corn flour, 200g of soybean meal, 80g of wheat bran, 30g of fish meal, 10g of bone meal, and 5g of salt are mixed to obtain basic pig feed powder. 900g of basic pig feed powder is pelleted by a pellet mill to obtain basic feed.

[0087] Experimental Example 1

[0088] In vitro experiments simulating the effects of the gastrointestinal tract on fat release from fat powder:

[0089] Add 10g of the fat powder from Examples 1-3 and Comparative Examples 1-3 to 200mL of 0.1M HCl (pH 2.0), respectively. Incubate at 37°C with shaking at 100rpm for 30min. Adjust the pH to 5.5, add 0.1% trypsin, and continue incubation at 37°C. Sample every hour, centrifuge at 8000rpm for 10min, collect the supernatant, and determine the fat content. Weigh 10.0g and place it in a 50mL test tube, add 10mL of hydrochloric acid. Place the test tube in a 70-80°C water bath and stir with a glass rod every 5-10min until the sample is completely digested, approximately 40-50min. Remove the test tube, add 10mL of ethanol, and mix. After cooling, transfer the mixture to a 100mL stoppered graduated cylinder. Wash the test tube several times with 25mL of ether, and pour the washed ether mixture into the graduated cylinder. After all the ether has been poured into the graduated cylinder, stopper it and shake for 1 minute. Carefully open the stopper to release the gas, then stopper it again and let it stand for 12 minutes. Carefully open the stopper again and rinse the stopper and the mouth of the cylinder with an equal volume mixture of petroleum ether and diethyl ether to remove any adhering fat. Let it stand for 10–20 minutes until the upper liquid is clear. Draw the supernatant into a pre-balanced conical flask, add 5 mL of diethyl ether to the stoppered graduated cylinder, shake, and let it stand. Draw the upper layer of ether back into the original conical flask. Evaporate the conical flask to dryness in a water bath, then dry it in an oven at 95–105°C for 2 hours. After cooling in a desiccator for 0.5 hours, weigh the flask and calculate the weight of the fat. The results are shown in Table 1 below.

[0090] Table 1: Release rate of fat from fat powder in simulated gastrointestinal tract

[0091]

[0092]

[0093] As shown in Table 1, the fat powder prepared by the methods in Examples 1-3 had a release rate of about 25% in the simulated gastrointestinal tract, while the fat powder prepared by the methods in Comparative Examples 1-3 had a release rate of more than 40% in the simulated gastrointestinal tract. In Comparative Examples 2 and 3, the release rate of the unencapsulated liposome fat powder exceeded 85%, indicating that the shell structure encapsulating the liposome fat powder has the effect of reducing the fat release rate and achieving the purpose of sustained fat release.

[0094] Experimental Example 2

[0095] Experiment on fat absorption in young piglets:

[0096] Twenty-one male piglets aged 21 days were divided into seven groups of three each. The seven groups were labeled as: Example 4, Example 5, Example 6, Comparative Example 4, Comparative Example 5, Comparative Example 6, and Comparative Example 7. Each piglet was fed 200g of the corresponding feed five times a day for seven days. Body weight was recorded before and after the experiment. Blood samples were collected intravenously before and after the experiment to measure serum triglyceride (TG), total cholesterol (TC), and high-density lipoprotein (HDL) levels, as shown in Table 2. After seven days of feeding, feces were collected, dried at 65℃, and the crude fat content was measured. 10.0g of dried feces was weighed and placed in a 50mL test tube, with 10mL of hydrochloric acid added. The test tube was placed in a 70-80℃ water bath, and stirred with a glass rod every 5-10 minutes until the sample was completely digested, approximately 40-50 minutes. The test tube was then removed, and 10mL of ethanol was added and mixed. After cooling, transfer the mixture to a 100mL stoppered graduated cylinder. Wash the test tubes with 25mL of diethyl ether in portions, pouring the washes into the graduated cylinder. After all the diethyl ether has been poured into the graduated cylinder, stopper the cylinder and shake for 1 minute. Carefully open the stopper to release the gas, then stopper the cylinder again and let it stand for 12 minutes. Carefully open the stopper again and rinse the stopper and the mouth of the cylinder with an equal volume mixture of petroleum ether and diethyl ether to remove any adhering fat. Let it stand for 10–20 minutes until the upper liquid is clear. Pour the supernatant into a pre-balanced Erlenmeyer flask, add 5mL of diethyl ether to the stoppered graduated cylinder, shake, and let it stand. Pour the upper layer of diethyl ether back into the original Erlenmeyer flask. Evaporate the Erlenmeyer flask to dryness in a water bath, then dry it in an oven at 95–105℃ for 2 hours. After cooling in a desiccator for 0.5 hours, weigh the flask and calculate the weight of the fat, as shown in Table 3.

[0097] Table 2: Serum biochemical indicators of fat absorption in young piglets

[0098]

[0099]

[0100] As shown in Table 2, the serum triglyceride, total cholesterol, and high-density lipoprotein concentrations of piglets fed with the feeds of Examples 4-6 were all higher than those of Comparative Examples 4-7, indicating that the feeds of Examples 4-6 can effectively promote the absorption of fat by piglets.

[0101] Table 3: Fat content in feces of young piglets

[0102]

[0103] As shown in Table 3, the fat content in the feces of young piglets fed the feeds of Examples 4-6 was lower than that in the feces of young piglets fed the feeds of Comparative Examples 4-6. This indicates that the feeds prepared in Examples 4-6 promote fat absorption in young animals and reduce the excretion of undigested fat in feces. The low fat content in the feces of young piglets fed the feed of Comparative Example 7 is because Comparative Example 7 only contained a basic feed without the addition of fat powder, resulting in a lower fat content in the basic feed.

[0104] Experimental Example 3

[0105] Intestinal lipase expression and activity verification experiment:

[0106] After slaughter, tissue was taken from the mid-section of the jejunum (1 m from the pylorus), and the inner wall mucosa was scraped off and divided into two parts. One part was placed in RNA preservation solution (-80℃) for gene expression detection; RT-PCR was used to detect the relative expression levels of AMPK and PPARα in the jejunal tissue (β-actin was the internal reference gene). -ΔΔCt Using β-actin as an internal reference, and with the expression level of Example 7 as 1, the relative expression levels of the other groups relative to Example 7 were calculated, and the results are as follows: Figure 3 and Figure 4 As shown; another portion was added to PBS buffer, homogenized, and stirred evenly for 1 hour; centrifuged at 10000 r / min for 20 min at 4℃, and the supernatant was filtered through a 0.45 μm filter membrane to obtain crude porcine pancreatic lipase (PL) solution. The activity of porcine pancreatic lipase (PL) in the intestinal contents (unit: U / g protein) was detected by ELISA (kit), and the results are shown in the figure. Figure 5 As shown.

[0107] like Figure 3 and Figure 4The data show that after feeding weaned piglets with the feeds prepared according to the methods in Examples 4-6, the expression level of AMPK in the piglets' intestines decreased, significantly lower than that in Comparative Examples 4-7; while after feeding weaned piglets with the feeds prepared according to the methods in Examples 4-6, the expression level of PPARα in the piglets' intestines significantly increased, significantly higher than that in Comparative Examples 4-7. This indicates that the feeds prepared according to the methods in Examples 4-6 can effectively promote lipid metabolism-related signaling pathways. Figure 5 As shown, after feeding weaned piglets with the feed prepared by the methods in Examples 4-6, the activity of pancreatic lipase in the piglets' intestines was significantly higher than that in Comparative Examples 4-7, indicating that the feed prepared by the methods in Examples 4-6 can increase the expression of lipase in the intestines and promote the absorption of fat in the intestines.

[0108] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a fat powder that promotes absorption in young animals, characterized in that, Includes the following steps: S1. Place 1-oleic acid-2-palmitic acid-3-linoleic acid triglyceride, soybean oil and egg yolk lecithin into a three-necked flask, purge with nitrogen, and stir in a water bath until a homogeneous oil phase is formed; in another beaker, add hydrogenated soybean lecithin and cholesterol, pour in chloroform, and stir until completely dissolved to obtain the membrane solution; S2. Pour the homogeneous oil phase into the membrane material solution and stir to form a mixture; transfer the mixture to a flask of a rotary evaporator and rotary evaporate until the chloroform is completely evaporated, forming a homogeneous lipid film on the inner wall of the flask; S3. Add PBS buffer to the flask in S2, and stir in a water bath for the second time until the lipid film is completely hydrated to form a crude liposome suspension; transfer the crude liposome suspension to an ultrasonic disruptor and sonicate to obtain nano-sized liposomes; spray dry the nano-sized liposomes to obtain liposome powder. S4. Ethyl cellulose and stearic acid are dissolved in anhydrous ethanol to obtain an ethyl cellulose-stearic acid solution. Liposome micropowder is added to a fluidized bed, sprayed with the ethyl cellulose-stearic acid solution, and dried for the first time to obtain bilayer microspheres. S5. Sodium butyrate, linolenic acid, and Lactobacillus reuteri were dispersed in deionized water, and gelatin and low-acyl gellan gum were added. The mixture was stirred and dissolved to obtain a shell gel. The bilayer microspheres were added to a fluidized bed, the shell gel was sprayed on, and the mixture was dried a second time to obtain a fat powder that promotes absorption in young animals.

2. The method for preparing a fat powder that promotes absorption in young animals according to claim 1, characterized in that, In S1, the mass ratio of 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride, soybean oil, and egg yolk lecithin is 50:(25-35):(5-15); the water bath stirring temperature is 45-55℃, and the stirring speed for the first water bath stirring is 250-350 rpm; the mass ratio of hydrogenated soybean lecithin, cholesterol, and 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride is (5-7):(1-3):50; the ratio of chloroform to 1-oleic acid-2-palmitoyl-3-linoleic acid triglyceride is (3.5-4.5) mL:1 g; and the stirring speed until complete dissolution is 150-250 rpm.

3. The method for preparing a fat powder that promotes absorption in young animals according to claim 1, characterized in that, In S2, the ratio of the homogeneous oil phase to the film material solution is 90g:(175~225)mL; the stirring speed for forming the mixture is 250~350rpm, and the time is 15~25min; the rotary evaporation temperature is 30~40℃, the vacuum degree is 0.07~0.09MPa, and the rotation speed is 50~70rpm.

4. The method for preparing a fat powder that promotes absorption in young animals according to claim 1, characterized in that, In S3, the molar concentration of the PBS buffer is 0.01–0.03 M, the pH is 7.2–7.6, and the ratio of the PBS buffer to the homogeneous oil phase in S2 is (900–1000) mL:90 g; the second water bath stirring temperature is 35–40 °C, and the rotation speed is 150–250 rpm; the ultrasonic power is 250–350 W, and the time is 8–12 min; the spray drying inlet air temperature is 55–65 °C, the outlet air temperature is 30–40 °C, and the feed rate is 8–12 mL / min.

5. The method for preparing a fat powder that promotes absorption in young animals according to claim 1, characterized in that, In S4, the ratio of ethyl cellulose, stearic acid, and anhydrous ethanol is 1 g:(0.8-1.2) g:(90-110) mL; the ratio of liposome micropowder to ethyl cellulose-stearic acid solution is 95 g:(450-550) mL; the fluidized bed inlet air temperature is 35-45°C; the spraying rate is 4-6 mL / min; and the first drying temperature is 35-45°C for 1.5-2.5 h.

6. The method for preparing a fat powder that promotes absorption in young animals according to claim 1, characterized in that, In S5, the ratio of sodium butyrate, linolenic acid, Lactobacillus reuteri, and deionized water is (0.4–0.6) g:(1.5–2.5) g:(0.5–1.5) g:500 mL; the ratio of gelatin, low-acyl gellan gum, and deionized water is (15–25) g:(4–6) g:500 mL; the fluidized bed inlet air temperature is 35–45 °C; the spraying rate is 6–10 mL / min; and the second drying temperature is 30–40 °C for 1.0–2.0 h.

7. The application of a fat powder prepared by the method according to any one of claims 1-6 that promotes absorption in young animals, characterized in that, The fat powder that promotes absorption by young animals is used as an additive in the feed of young animals.

8. The application of a fat powder that promotes absorption in young animals according to claim 7, characterized in that, The feed for young animals is a basic feed powder for young animals, and the mass ratio of the feed additives for young animals to the basic feed powder for young animals is 1:(8-10).

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