Preparation and application of composite polypeptide growth-promoting supplement liquid for livestock and poultry

Abstract

The invention relates to the technical field of agricultural science, and particularly discloses preparation and application of a composite polypeptide growth-promoting supplement liquid for livestock and poultry. The composition comprises a plurality of active polypeptides such as recombinant human epidermal growth factors, trefoil peptides, insulin-like growth factors and the like, vitamins A, E, C and B, and is supplemented with recombinant collagen peptides, marine source acidic polysaccharides and a composite carrier consisting of hydroxypropyl-beta-cyclodextrin, trehalose and sodium hyaluronate. The preparation method comprises the following steps: respectively carrying out independent shunt pretreatment on the polypeptide component and the vitamin component, integrating through a programmed gradient compounding process and a dynamic pH regulation process, and finally carrying out fine filtration and sterile filling to obtain the product. The composition can be used for preparing a feed or drinking water additive for promoting the growth of livestock and poultry, maintaining intestinal health and improving the feed utilization rate, and has the advantages of stable components and good activity maintaining effect; according to the preparation method, the long-acting stability of the composite system is realized by optimizing the process sequence and conditions.

Description

Technical Field

[0001] This application relates to the field of agricultural science and technology, and more specifically, to the preparation and application of a compound polypeptide growth-promoting supplement for livestock and poultry. Background Technology

[0002] In livestock and poultry farming, growth-promoting fluid supplements are widely used to meet the growth and development needs of livestock and poultry and maintain stable intestinal physiological function. These products typically integrate multiple active ingredients such as peptides, vitamins, and polysaccharides. Through the synergistic physiological effects of these ingredients, they help improve livestock and poultry production performance and ensure farming efficiency. Their core application value lies in allowing the active ingredients to fully exert their effects and provide the necessary physiological support for livestock and poultry growth.

[0003] Traditional preparations of growth-promoting fluids for livestock and poultry often employ simple mixing methods, failing to address the interactions between different types of active ingredients and lacking systematic process design to ensure component stability. This leads to easy interference between different active ingredients, insufficient component stability, and consequently, the inability of each component to fully exert its physiological effects, thus affecting the actual application efficacy of the product. Summary of the Invention

[0004] To address the problems of active ingredients in existing growth-promoting fluids for livestock and poultry easily interfering with each other, lacking stability, and failing to fully exert their physiological effects synergistically, this application provides a preparation and application of a compound polypeptide growth-promoting fluid for livestock and poultry.

[0005] In a first aspect, this application provides a method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry, employing the following technical solution: A method for preparing a compound polypeptide growth-promoting rehydration solution for livestock and poultry, comprising the following components by weight: 0.8-1 parts recombinant human epidermal growth factor, 1-1.2 parts trefoil peptide, 0.6-0.8 parts insulin-like growth factor, 20-30 parts vitamin A palmitate, 200-220 parts vitamin B complex, 300-340 parts magnesium L-ascorbic acid-2-phosphate, 80-120 parts D-α-tocopherol succinate, 400-480 parts recombinant collagen peptide, 100-120 parts marine-derived acidic polysaccharide, 380-460 parts composite carrier, 120-180 parts pH adjuster, and 10-12 parts stabilizer.

[0006] By employing the above technical solutions, recombinant human epidermal growth factor, trefoil peptide, insulin-like growth factor, and recombinant collagen peptide constitute a specific complex polypeptide system. Each of the four polypeptides exerts a targeted technical effect based on its own structural characteristics: recombinant human epidermal growth factor can act on intestinal epithelial cells of livestock and poultry; trefoil peptide can participate in intestinal mucosal barrier-related physiological processes; insulin-like growth factor can regulate metabolic pathways; and recombinant collagen peptide provides structural support for intestinal mucosal repair. The four peptides achieve synergistic effects through functional complementarity. Marine-derived acidic polysaccharides, with their acidic groups and molecular structure, can assist in the adsorption and dispersion of active ingredients in the intestine, while also providing a dispersion medium for lipid-soluble components. The complex carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate. These three components form multiple protective mechanisms through structural complementarity, providing a stable loading and delivery basis for the active ingredients. Vitamin A palmitate, L-ascorbic acid-2-phosphate magnesium, and D-α-tocopherol succinate, as stabilized vitamin derivatives, possess stronger antioxidant and decomposition capabilities compared to ordinary vitamins, maintaining their activity during storage and application. The vitamin B complex provides essential coenzyme support for livestock and poultry metabolism, ensuring smooth metabolic pathways. pH adjusters regulate and maintain the system's acidity and alkalinity, providing a suitable environment for each component. Stabilizers inhibit the degradation and polymerization of active ingredients from the perspectives of structural protection and antioxidant properties. Through precise formulation and functional synergy, these components construct a structurally stable and mutually compatible composite system.

[0007] Preferably, the vitamin B complex comprises thiamine hydrochloride, sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate in a mass ratio of (0.8-1.2):(1.0-1.4):(0.8-1.2):(1.8-2.2):(1.3-1.7); the composite carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate, wherein the mass ratio of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate is 5:3:1 to 8:5:2.

[0008] By employing the above technical solution, thiamine hydrochloride, riboflavin-5'-phosphate sodium, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate are all active forms of key coenzymes in livestock and poultry metabolism. Thiamine hydrochloride participates in carbohydrate catabolism, riboflavin-5'-phosphate sodium mediates redox reactions in vivo, pyridoxine hydrochloride regulates amino acid metabolism and protein synthesis, nicotinamide is an important component of energy metabolism-related enzymes, and D-calcium pantothenate participates in fatty acid synthesis and metabolic pathways. The five components are combined in a specific mass ratio to ensure functional adaptation in metabolic pathways, avoiding metabolic imbalances caused by excess or deficiency of a single component. By synergistically providing the necessary coenzyme support for metabolism, the smooth progress of physiological metabolic processes in livestock and poultry is guaranteed. Hydroxypropyl-β-cyclodextrin has a hydrophobic cavity structure, which can contain hydrophobic active ingredients through inclusion. The hydroxyl groups in trehalose molecules can form hydrogen bonds with protein molecules, inhibiting protein spatial structure denaturation. Low molecular weight sodium hyaluronate has good hydrophilicity and biocompatibility, which can improve the interaction between the carrier and the intestinal mucosa. When the three components are combined in a mass ratio of 5:3:1 to 8:5:2, their inclusion, hydrogen bond stabilization, and mucosal compatibility optimization work synergistically to resist the influence of the external environment on the active ingredients, thus helping the ingredients to function effectively in the intestine and achieving the dual functions of stabilizing and delivering the active ingredients.

[0009] Preferably, the trifoliate peptide is selected from at least one of TFF1, TFF2, or TFF3; the insulin-like growth factor is selected from IGF-1 or IGF-2; the pH adjuster is selected from at least one of hydrochloric acid, sodium hydroxide, citric acid, sodium citrate, sodium dihydrogen phosphate, or disodium hydrogen phosphate; the stabilizer is selected from at least one of trehalose, L-methionine, sorbitol, mannitol, or ascorbyl palmitate; and the marine-derived acidic polysaccharide is selected from at least one of alginate oligosaccharide, carrageenan oligosaccharide, or fucoidan sulfate oligosaccharide.

[0010] By employing the above technical solutions, TFF1, TFF2, and TFF3, all belonging to the trefoil factor family, each possess specific amino acid sequences and spatial structures. They play targeted technical roles in intestinal mucosal protection, damage repair, and epithelial cell homeostasis maintenance, respectively. Selecting at least one of them can adapt to the needs of different intestinal physiological scenarios, strengthening mucosal barrier function through interactions with intestinal mucosal-related sites. IGF-1 and IGF-2, as growth-related factors, undertake specific functions in pathways such as protein synthesis regulation and cell proliferation and differentiation, adapting to the physiological and metabolic needs of livestock and poultry at different growth stages, participating in the growth and development process of livestock and poultry by mediating growth-related signal transduction. Hydrochloric acid and sodium hydroxide can directly regulate the pH of the system. Citric acid and sodium citrate, and sodium dihydrogen phosphate and disodium hydrogen phosphate, respectively constitute buffer systems. Buffer systems can inhibit drastic pH fluctuations caused by external factors. Different types of pH adjusters can be flexibly selected according to the initial state and stability requirements of the system. Trehalose stabilizes protein spatial structure through hydrogen bonding; L-methionine scavenge free radicals and inhibits oxidation; sorbitol and mannitol regulate osmotic pressure and reduce component loss; ascorbyl palmitate combines antioxidant and lipid-soluble component stabilization functions, resulting in complementary effects among different stabilizers. Alginate oligosaccharides, carrageenan oligosaccharides, and fucoidan sulfate oligosaccharides all contain numerous acidic functional groups, exhibiting good water solubility and dispersibility. They can adsorb onto the intestinal mucosa and provide a dispersion medium for lipid-soluble components, promoting uniform distribution and effective adsorption of active ingredients in the intestine, thus facilitating their physiological functions.

[0011] Secondly, this application provides a preparation process for a compound polypeptide growth-promoting supplement for livestock and poultry, employing the following technical solution: A preparation process for a compound polypeptide growth-promoting supplement for livestock and poultry includes the following steps: S1. Construction and activation of the structured carrier: Hydroxypropyl-β-cyclodextrin and trehalose were dissolved in phosphate buffer to prepare solution A; low molecular weight sodium hyaluronate was dissolved in the same buffer to prepare solution B; under continuous stirring and low temperature conditions, solution B was slowly added dropwise to solution A, and stirring was continued after the addition was completed to obtain the structured composite carrier solution. S2. Sequential embedding and stabilization of peptide components: Recombinant collagen peptides are dissolved in the composite carrier solution prepared in part of step S1 to form solution C; while maintaining the system at low temperature, recombinant human epidermal growth factor is first added to solution C, stirred to dissolve and allowed to stand; then, trefoil peptide and insulin-like growth factor are added sequentially, and each peptide is gently stirred at an appropriate speed to ensure full dispersion, thus obtaining the peptide-embedded composite solution. S3. Construction of the microenvironment for the vitamin complex system: Marine-derived acidic polysaccharides were dissolved in deionized water to prepare acidic polysaccharide solution D; vitamin A palmitate and D-α-tocopherol succinate were mixed with acidic polysaccharide solution D and emulsified in a water bath to obtain primary fat-soluble microemulsion E; vitamin B complex and magnesium L-ascorbic acid-2-phosphate were dissolved in the remaining complex carrier solution to obtain aqueous vitamin solution F; under room temperature and nitrogen atmosphere, primary fat-soluble microemulsion E was slowly injected into aqueous vitamin solution F and homogenized and circulated to obtain vitamin complex emulsion G; S4. Programmed gradient compounding and phase integration: The peptide-loaded composite solution obtained in step S2 is transferred to a reaction vessel and stirred. The vitamin composite emulsion G obtained in step S3 is pumped into the reaction vessel. After pumping, the temperature is programmed and the integration reaction continues at the temperature after the temperature is increased to obtain the integrated solution. S5. Dynamic pH control and terminal stabilization: Add a pH adjuster slowly to the integrated solution obtained in step S4 while stirring to precisely adjust the pH value of the system step by step, and then add a stabilizer. S6. Multi-mode terminal refining and aseptic collection: The liquid processed in step S5 is subjected to graded filtration to obtain refined replenishment liquid, which is then collected in an aseptic receiving tank. S7. Aseptic filling and finished product handling: The refined replenishing solution is filled into pre-sterilized light-proof containers under aseptic conditions; the atmosphere is immediately replaced after filling, and the container is sealed and stored in the dark under refrigeration conditions.

[0012] By employing the above technical solution, the preparation process utilizes a combination of pretreatment and precise control of peptides and vitamins to construct a stable system adapted for multi-component synergy: First, hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate are dissolved separately in phosphate buffer. Under low temperature conditions, these are slowly added dropwise with continuous stirring, allowing the three carrier components to fully interact and form a carrier system with a network inclusion structure, providing a stable matrix for subsequent loading of active ingredients. Peptide components are added to the carrier solution in a specific order, with low temperature and gentle stirring reducing interactions between peptide molecules. The static setting process helps the peptides and carriers form a stable bond, preventing damage to the active structure. The vitamin system employs a phase-separation treatment method. Fat-soluble vitamins are emulsified to form microemulsions through the dispersion effect of marine-derived acidic polysaccharides, while water-soluble vitamins dissolve in the remaining carrier solution. Then, homogenous circulation under nitrogen protection (to prevent oxidation of fat-soluble vitamins) achieves stable coexistence of the oil and water dispersed phases, ensuring the dispersion stability of the vitamin components. The peptide-encapsulated solution and vitamin emulsion are slowly pumped in and temperature-programmed to achieve gradient blending. Continuous stirring promotes full integration of the system. Stepwise pH adjustment, using a stable range followed by precise fine-tuning, provides a suitable acid-base environment for each component. The subsequent addition of stabilizers further inhibits component degradation. Staged filtration removes impurities and aggregated particles through the sequential action of filter cartridges with different pore sizes. Aseptic filling, combined with nitrogen atmosphere replacement and light-proof refrigerated storage, reduces the impact of external environmental contamination and oxidation on the product. The entire process, through precise control of each step, achieves the stable coexistence and effective retention of multiple components.

[0013] Preferably, in step S1, the pH value of the phosphate buffer solution is 6.8 to 7.2; the concentration of solution A is 10-20% w / v, and the concentration of solution B is 2-5% w / v; the low temperature condition is 4-8℃; the stirring speed is 100-300 rpm, and the stirring time is 30-60 min.

[0014] By employing the above technical solution, the phosphate buffer solution is selected within a pH range of 6.8 to 7.2 to match the solubility characteristics of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate, preventing precipitation or structural changes due to unsuitable acid-base environments and providing a stable medium for the interaction of the carrier components. Solution A is set at a concentration of 10-20% w / v to ensure that hydroxypropyl-β-cyclodextrin and trehalose are fully dissolved and form a matrix system of suitable concentration. Solution B uses a concentration of 2-5% w / v to match the water solubility characteristics of low molecular weight sodium hyaluronate, preventing molecular aggregation due to excessive concentration and ensuring uniform contact with solution A during dropwise addition. The low temperature condition of 4-8℃ can inhibit excessive movement of carrier component molecules, reduce non-specific aggregation, and promote the formation of specific interactions such as hydrogen bonds and hydrophobic interactions among the three components, thus facilitating the orderly construction of the structured carrier. A stirring speed of 100-300 rpm can achieve rapid mixing of solution B and solution A while avoiding excessive shear force that could damage the component structure. A stirring time of 30-60 min ensures that the three carrier components are in full contact and can penetrate each other, completing the activation process of the structured carrier and forming a network structure with stable inclusion capacity, which provides a basis for the specific loading of active ingredients.

[0015] Preferably, in step S2, the concentration of solution C is 8.0-12.0% w / v; the low temperature condition is 2.0-6.0℃; the standing time is 15-17 min; the gentle stirring speed is 100-200 rpm; and the stirring time after each polypeptide is added is 10-12 min.

[0016] By adopting the above technical solution and setting the solution concentration (C) to 8.0-12.0% w / v, the recombinant collagen peptides are fully dissolved in the carrier solution, while sufficient binding sites are reserved for subsequent peptide loading. This avoids peptide aggregation due to excessively high concentrations or the interaction efficiency between the carrier and peptides being affected by excessively low concentrations. The low temperature (2.0-6.0℃) further reduces the thermal motion intensity of peptide molecules, minimizing the risk of spatial structural denaturation of recombinant human epidermal growth factor, trefoil peptide, and insulin-like growth factor, thus providing an environment conducive to peptide activity retention. A 15-17 min settling period helps the recombinant collagen peptides form a stable binding system with the carrier, constructing a base structure that provides specific binding sites for subsequent peptides. A gentle stirring speed of 100-200 rpm ensures uniform mixing of the subsequently added trefoil peptide, insulin-like growth factor, and solution C without shearing damage to the peptide structure. A stirring time of 10-12 min after each peptide addition ensures that each peptide is fully dispersed and forms a targeted binding with the carrier system, avoiding mutual interference caused by the simultaneous mixing of multiple peptides.

[0017] Preferably, in step S3, the concentration of the acidic polysaccharide solution D is 3.0% w / v to 6.0% w / v; the temperature of the water bath is 45-55℃; the emulsification speed is 8000-12000 rpm; the emulsification time is 3-5 min; the pressure of the homogenization cycle is 80-120 MPa; and the number of cycles is 2-4.

[0018] By adopting the above technical solution, an acidic polysaccharide solution D with a concentration of 3.0% w / v to 6.0% w / v is adapted to the water solubility and dispersion characteristics of marine-derived acidic polysaccharides. This ensures that the polysaccharides are fully dissolved to form a homogeneous solution, while also providing sufficient acidic functional groups to interact with fat-soluble vitamins, thus providing a stable medium for the dispersion of fat-soluble components. A water bath temperature of 45-55℃ can reduce the viscosity of fat-soluble vitamins without damaging their activity, making them easier to disperse into tiny droplets and creating suitable conditions for the emulsification process. A high emulsification speed of 8000-12000 rpm generates sufficient shear force to disperse the mixture of fat-soluble vitamins and acidic polysaccharide solution D into uniform, submicron-sized droplets. An emulsification time of 3-5 minutes ensures that the droplet size reaches a stable state, forming a structurally uniform primary fat-soluble microemulsion E. A homogenization cycle pressure of 80-120 MPa can further refine the microemulsion droplet size and improve droplet stability. 2-4 cycles can achieve sufficient droplet refinement while avoiding over-processing that could damage the component structure. Ultimately, the primary fat-soluble microemulsion E and the aqueous vitamin solution F are uniformly fused to form a stable vitamin complex emulsion G.

[0019] Preferably, in step S4, the stirring temperature is 8-12℃; the stirring speed is 150-250 rpm; the pumping is completed uniformly within 30-50 minutes; the programmed heating rate is 0.3-0.5℃ / min, and the temperature is increased to 22-26℃; the rotation speed of the integrated reaction is 300-400 rpm, and the reaction time is 90-150 minutes.

[0020] By employing the above technical solution, a stirring temperature of 8-12℃ can maintain the structural stability of the peptide components in the peptide-encapsulated composite solution while avoiding excessively low temperatures that could lead to increased viscosity and mixing difficulties in the two emulsion systems. This provides a suitable temperature environment for the initial fusion of the peptide-encapsulated composite solution and the vitamin complex emulsion G. A stirring speed of 150-250 rpm allows for gradual mixing of the two systems without damaging the binding structure between the peptide and the carrier or the stability of the vitamin microemulsion droplets, laying the foundation for subsequent deep integration. A uniform pumping rate of vitamin complex emulsion G over 30-50 minutes avoids localized high concentrations and phase separation caused by rapid contact between the two systems, allowing the peptide-encapsulated composite solution and vitamin complex emulsion G to gradually penetrate and adapt to each other during stirring. A slow programmed heating rate of 0.3-0.5℃ / min reduces the impact of sudden temperature changes on the structure of the active ingredients. Gradually increasing the system temperature to 22-26℃ within this temperature range promotes specific binding between the peptides, vitamins, and the carrier through hydrogen bonding and hydrophobic interactions, while preventing excessively high temperatures from causing activity degradation. The integration reaction speed of 300-400 rpm provides sufficient hybrid power to facilitate the deep integration of the two systems, and the reaction time of 90-150 min ensures that the components are in full contact and interact with each other, ultimately forming a homogeneous integrated liquid without stratification or particle precipitation.

[0021] Preferably, in step S5, the stepwise precise adjustment specifically involves: first adjusting the pH to 6.2 to 6.8 and maintaining it for 15 minutes, then fine-tuning it to 6.5 to 7.0 and maintaining it for 5-10 minutes; in step S6, the graded filtration specifically involves: first pre-filtration through a 0.45μm pore size filter element, then sequentially passing through sterile filter elements with 0.22μm and 0.1μm pore sizes; in step S7, the light-proof container is a pre-sterilized brown type III glass bottle; the gas used for atmosphere replacement is nitrogen, wherein the headspace oxygen content is less than 2.0% v / v; and the refrigeration conditions are 2-8℃.

[0022] By employing the above technical solution, when precisely adjusting the pH step by step, the system is first adjusted to 6.2 to 6.8 and held for 15 minutes. This range is suitable for the stable existence requirements of peptides, vitamins, and carrier components, giving each component sufficient time to adapt to the acidic or alkaline environment and avoiding structural denaturation or aggregation caused by sudden pH changes. Then, it is finely adjusted to 6.5 to 7.0 and held for 5-10 minutes to further precisely match the optimal stable pH range of all components in the system. Through two-stage adjustment and temperature maintenance, the pH of the system is precisely stabilized, with pH fluctuations controlled within ±0.1, ensuring the long-term coexistence of all components. During the staged filtration process, the 0.45μm pore size filter cartridge first intercepts larger-sized impurities and a small amount of aggregated particles in the system, preventing subsequent fine-pore size filter cartridges from clogging and ensuring filtration efficiency. Subsequently, it passes through sterilization-grade filter cartridges with 0.22μm and 0.1μm pore sizes in sequence, gradually intercepting tiny impurities, bacteria, and microorganisms, achieving deep purification and sterilization of the refined replenishment solution, ensuring that the microbial content meets the sterile standards for feed additives and reducing the risk of contamination. Pre-sterilized brown Type III glass bottles are used as light-proof containers. The brown material blocks ultraviolet light penetration, preventing the active ingredients from being oxidized and degraded by light. The sterilization process eliminates the risk of contamination from the container itself. Nitrogen is used for atmosphere replacement to keep the headspace oxygen content below 2.0% v / v, reducing the damage of oxygen to easily oxidized components in the system. Refrigeration conditions of 2-8℃ can reduce the intensity of thermal motion of component molecules and inhibit the degradation reaction rate. Combined with sealing treatment, the stability of the product during storage is guaranteed from multiple dimensions such as light, oxygen, and temperature. There is no obvious component degradation or color change during storage.

[0023] Thirdly, this application provides an application of a compound polypeptide growth-promoting fluid for livestock and poultry, employing the following technical solution: An application of a compound polypeptide growth-promoting supplement for livestock and poultry, used to prepare feed additives or drinking water additives that promote livestock and poultry growth, maintain intestinal health, and improve feed utilization.

[0024] By adopting the above technical solution, this rehydration solution, relying on its uniform and stable liquid form, can be directly added to livestock and poultry drinking water in an appropriate ratio, or mixed with feed ingredients through a uniform mixing process to form a feed additive, adapting to the conventional intake scenarios in livestock and poultry farming. The complex polypeptides, vitamin complexes, marine-derived acidic polysaccharides, and other components in the rehydration solution, after being ingested by livestock and poultry, can help maintain intestinal physiological homeostasis and digest and absorb nutrients by interacting with the intestinal mucosa and participating in metabolic pathways in the body. Its composition and the stability guaranteed by the process enable the rehydration solution to be used as a feed additive or drinking water additive, providing support for relevant technical needs in livestock and poultry farming.

[0025] In summary, this application has the following beneficial effects: 1. Because this application uses a complex polypeptide combination of recombinant human epidermal growth factor, trefoil peptide, insulin-like growth factor and recombinant collagen peptide, combined with marine-derived acidic polysaccharide and complex carrier, and through preparation processes such as pretreatment, program integration and dynamic pH control, the components have good stability and can fully exert their related physiological effects due to the synergistic effect of each component and the process avoids interference between components.

[0026] 2. In this application, a composite carrier composed of hydroxypropyl-β-cyclodextrin, trehalose and low molecular weight sodium hyaluronate in a specific ratio is preferred, and the structured carrier is constructed by controlling the pH value of phosphate buffer and stirring parameters. Since this carrier system can effectively protect the active ingredients, the retention rate of active ingredients is improved.

[0027] 3. The method of this application processes polypeptide components by sequentially embedding polypeptides, allowing them to stand at low temperatures, and gently stirring them. At the same time, vitamins are emulsified and homogenized to construct a microenvironment, thus achieving uniform dispersion of each active ingredient.

[0028] 4. This application achieves high product purity and stable performance during storage by precisely adjusting the pH value step by step and maintaining it for a specific time, combined with multi-stage sterilization filtration, nitrogen atmosphere replacement, and aseptic filling. This stabilizes the acidity and alkalinity of the system, removes impurities, and prevents contamination.

[0029] 5. The rehydration solution of this application uses stabilized vitamin derivatives, combined with specific pH adjusters and stabilizers, and is suitable for use as a feed additive or drinking water additive. Because the components are suitable for the ingestion scenarios of livestock and poultry and are not easily oxidized and degraded, it has achieved good compatibility and can play a related role for a long time. Attached Figure Description

[0030] Figure 1 This is a flowchart of a method for preparing a compound polypeptide growth-promoting fluid for livestock and poultry, as provided in this application. Detailed Implementation

[0031] The present application will be further described in detail below with reference to embodiments and comparative examples. Unless otherwise specified, the experimental methods used below are conventional methods. Unless otherwise specified, the materials, reagents, methods and instruments used are all conventional materials, reagents, methods and instruments in the art, which can be obtained by those skilled in the art through commercial channels or prepared according to literature methods.

[0032] Technical concept: In related technologies, growth-promoting fluid supplements for livestock and poultry suffer from problems such as the inability of active ingredients to work synergistically and insufficient stability. The core reason lies in the lack of targeted design and systematic support for process control in ingredient formulation: at the ingredient level, a functionally complementary complex system has not been formed, common vitamins are easily oxidized and degraded, and a single carrier is difficult to achieve stable loading of multiple types of active ingredients; at the process level, simple mixing mode is often used, without controlling the interaction between different types of ingredients such as peptides and vitamins, and lacking operations such as pre-treatment and precise parameter control, which leads to easy aggregation, denaturation or mutual interference between ingredients, ultimately affecting the application effect of the product.

[0033] This technical solution addresses the aforementioned issues through synergistic innovation in precise component compatibility and process system design. In terms of components, a complex peptide system is constructed, combined with stabilized vitamin derivatives, and paired with a composite carrier composed of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate, along with marine-derived acidic polysaccharides, forming a functionally complementary and structurally compatible component system. In terms of process, a two-pronged pretreatment strategy is adopted for peptides and vitamins. Through a series of operations including structured carrier construction, sequential peptide embedding, vitamin emulsification and homogenization, programmed temperature gradient compounding, and stepwise precise pH control, combined with graded filtration and aseptic storage processes, interference between components is suppressed from both component compatibility and process control perspectives, protecting the structural stability of active ingredients and ultimately achieving synergistic coexistence and effective action of multiple components.

[0034] Example 1 This application provides a compound polypeptide growth-promoting supplement for livestock and poultry, comprising the following components by weight: 0.8 parts recombinant human epidermal growth factor, 1.0 part trefoil peptide, 0.6 parts insulin-like growth factor, 20 parts vitamin A palmitate, 200 parts vitamin B complex, 300 parts magnesium L-ascorbic acid-2-phosphate, 80 parts D-α-tocopherol succinate, 400 parts recombinant collagen peptide, 100 parts marine-derived acidic polysaccharide, 380 parts compound carrier, 120 parts pH adjuster, and 10 parts stabilizer. The vitamin B complex contains thiamine hydrochloride, sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate in a mass ratio of 0.8:1.0:0.8:1.8:1.3. The composite carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose and low molecular weight sodium hyaluronate in a mass ratio of 5:3:1. Among them, the trifolin peptide is TFF2, the insulin-like growth factor is IGF-1, the pH adjuster is a mixture of citric acid and sodium citrate in a mass ratio of 1:1, the stabilizer is a mixture of trehalose and L-methionine in a mass ratio of 10:1, and the marine-derived acidic polysaccharide is alginate oligosaccharide.

[0035] The preparation method of the above-mentioned compound polypeptide growth-promoting rehydration solution for livestock and poultry includes the following steps: S1. Construction and activation of the structured carrier: Hydroxypropyl-β-cyclodextrin and trehalose were dissolved in phosphate buffer to prepare solution A; low molecular weight sodium hyaluronate was dissolved in the same buffer to prepare solution B; under continuous stirring and low temperature conditions, solution B was slowly added dropwise to solution A, and stirring was continued after the addition was completed to obtain the structured composite carrier solution. The phosphate buffer solution has a pH of 6.8; the concentration of solution A is 10% w / v and the concentration of solution B is 2% w / v; the low temperature condition is 4°C; the stirring speed is 100 rpm and the stirring time is 30 min.

[0036] S2. Sequential embedding and stabilization of peptide components: Recombinant collagen peptides are dissolved in the composite carrier solution prepared in part of step S1 to form solution C; while maintaining the system at low temperature, recombinant human epidermal growth factor is first added to solution C, stirred to dissolve and allowed to stand; then, trefoil peptide and insulin-like growth factor are added sequentially, and each peptide is gently stirred at an appropriate speed to ensure full dispersion, thus obtaining the peptide-embedded composite solution. The concentration of solution C is 8.0% w / v; the low temperature condition is 2.0℃; the standing time is 15 min; the gentle stirring speed is 100 rpm; and the stirring time after each polypeptide is added is 10 min.

[0037] S3. Construction of the microenvironment for the vitamin complex system: Marine-derived acidic polysaccharides were dissolved in deionized water to prepare acidic polysaccharide solution D; vitamin A palmitate and D-α-tocopherol succinate were mixed with acidic polysaccharide solution D and emulsified in a water bath to obtain primary fat-soluble microemulsion E; vitamin B complex and magnesium L-ascorbic acid-2-phosphate were dissolved in the remaining complex carrier solution to obtain aqueous vitamin solution F; under room temperature and nitrogen atmosphere, primary fat-soluble microemulsion E was slowly injected into aqueous vitamin solution F and homogenized and circulated to obtain vitamin complex emulsion G; The concentration of acidic polysaccharide solution D is 3.0% w / v; the temperature of the water bath is 45℃; the emulsification speed is 8000 rpm; the emulsification time is 3 min; the pressure of the homogenization cycle is 80 MPa; and the number of cycles is 2.

[0038] S4. Programmed gradient compounding and phase integration: The peptide-loaded composite solution obtained in step S2 is transferred to a reaction vessel and stirred. The vitamin composite emulsion G obtained in step S3 is pumped into the reaction vessel. After pumping, the temperature is programmed and the integration reaction continues at the temperature after the temperature is increased to obtain the integrated solution. The stirring temperature is 8℃; the stirring speed is 150 rpm; the pumping is completed at a constant speed within 30 minutes; the programmed heating rate is 0.3℃ / min, heating to 22℃; the rotation speed of the integrated reaction is 300 rpm, and the reaction time is 90 minutes.

[0039] S5. Dynamic pH control and terminal stabilization: Add a pH adjuster slowly to the integrated solution obtained in step S4 while stirring to precisely adjust the pH value of the system step by step, and then add a stabilizer. Specifically, the stepwise precise adjustment involves first adjusting the pH to 6.2 and holding it for 15 minutes, then fine-tuning it to 6.5 and holding it for 5 minutes.

[0040] S6. Multi-mode terminal refining and aseptic collection: The liquid processed in step S5 is subjected to graded filtration to obtain refined replenishment liquid, which is then collected in an aseptic receiving tank. Specifically, the graded filtration process involves: first, pre-filtration through a 0.45μm pore size filter element, followed by sequential passing through sterilization-grade filter elements with 0.22μm and 0.1μm pore sizes.

[0041] S7. Aseptic filling and finished product handling: The refined replenishing solution is filled into pre-sterilized light-proof containers under aseptic conditions; the atmosphere is immediately replaced after filling, and the container is sealed and stored in the dark under refrigeration conditions. The light-proof container is a pre-sterilized brown type III glass bottle; the gas used for atmosphere replacement is nitrogen, with a headspace oxygen content of less than 2.0% v / v; and the refrigeration condition is 2°C.

[0042] Example 2 This application provides a compound polypeptide growth-promoting supplement for livestock and poultry, comprising the following components by weight: 0.9 parts recombinant human epidermal growth factor, 1.1 parts trefoil peptide, 0.7 parts insulin-like growth factor, 25 parts vitamin A palmitate, 210 parts vitamin B complex, 320 parts magnesium L-ascorbic acid-2-phosphate, 100 parts D-α-tocopherol succinate, 440 parts recombinant collagen peptide, 110 parts marine-derived acidic polysaccharide, 420 parts compound carrier, 150 parts pH adjuster, and 11 parts stabilizer. The vitamin B complex contains thiamine hydrochloride, sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate in a mass ratio of 1.0:1.2:1.0:2.0:1.5. The composite carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose and low molecular weight sodium hyaluronate in a mass ratio of 6.5:4:1.5. Among them, the trifolin peptide is a mixture of TFF1 and TFF2 in a mass ratio of 1:1; the insulin-like growth factor is a mixture of IGF-1 and IGF-2 in a mass ratio of 1:1; the pH adjuster is a mixture of sodium dihydrogen phosphate and disodium hydrogen phosphate in a mass ratio of 2:1; the stabilizer is a mixture of trehalose and L-methionine in a mass ratio of 15:1; and the marine-derived acidic polysaccharide is a mixture of carrageenan oligosaccharide and fucoidan sulfate oligosaccharide in a mass ratio of 1:1.

[0043] The preparation method of the above-mentioned compound polypeptide growth-promoting rehydration solution for livestock and poultry includes the following steps: S1. Construction and activation of the structured carrier: Hydroxypropyl-β-cyclodextrin and trehalose were dissolved in phosphate buffer to prepare solution A; low molecular weight sodium hyaluronate was dissolved in the same buffer to prepare solution B; under continuous stirring and low temperature conditions, solution B was slowly added dropwise to solution A, and stirring was continued after the addition was completed to obtain the structured composite carrier solution. The phosphate buffer solution has a pH of 7.0; the concentration of solution A is 15% w / v, and the concentration of solution B is 3.5% w / v; the low temperature condition is 6°C; the stirring speed is 200 rpm, and the stirring time is 45 min.

[0044] S2. Sequential embedding and stabilization of peptide components: Recombinant collagen peptides are dissolved in the composite carrier solution prepared in part of step S1 to form solution C; while maintaining the system at low temperature, recombinant human epidermal growth factor is first added to solution C, stirred to dissolve and allowed to stand; then, trefoil peptide and insulin-like growth factor are added sequentially, and each peptide is gently stirred at an appropriate speed to ensure full dispersion, thus obtaining the peptide-embedded composite solution. The concentration of solution C is 10.0% w / v; the low temperature condition is 4.0℃; the standing time is 16 min; the gentle stirring speed is 150 rpm; and the stirring time after each polypeptide is added is 11 min.

[0045] S3. Construction of the microenvironment for the vitamin complex system: Marine-derived acidic polysaccharides were dissolved in deionized water to prepare acidic polysaccharide solution D; vitamin A palmitate and D-α-tocopherol succinate were mixed with acidic polysaccharide solution D and emulsified in a water bath to obtain primary fat-soluble microemulsion E; vitamin B complex and magnesium L-ascorbic acid-2-phosphate were dissolved in the remaining complex carrier solution to obtain aqueous vitamin solution F; under room temperature and nitrogen atmosphere, primary fat-soluble microemulsion E was slowly injected into aqueous vitamin solution F and homogenized and circulated to obtain vitamin complex emulsion G; The concentration of acidic polysaccharide solution D is 4.5% w / v; the temperature of the water bath is 50℃; the emulsification speed is 10000 rpm; the emulsification time is 4 min; the pressure of the homogenization cycle is 100 MPa; and the number of cycles is 3.

[0046] S4. Programmed gradient compounding and phase integration: The peptide-loaded composite solution obtained in step S2 is transferred to a reaction vessel and stirred. The vitamin composite emulsion G obtained in step S3 is pumped into the reaction vessel. After pumping, the temperature is programmed and the integration reaction continues at the temperature after the temperature is increased to obtain the integrated solution. The stirring temperature is 10℃; the stirring speed is 200 rpm; the pumping is completed at a constant speed within 40 minutes; the programmed heating rate is 0.4℃ / min, heating to 24℃; the rotation speed of the integrated reaction is 350 rpm, and the reaction time is 120 minutes.

[0047] S5. Dynamic pH control and terminal stabilization: Add a pH adjuster slowly to the integrated solution obtained in step S4 while stirring to precisely adjust the pH value of the system step by step, and then add a stabilizer. Specifically, the stepwise precise adjustment involves first adjusting the pH to 6.5 and holding it for 15 minutes, then fine-tuning it to 6.7 and holding it for 7.5 minutes.

[0048] S6. Multi-mode terminal refining and aseptic collection: The liquid processed in step S5 is subjected to graded filtration to obtain refined replenishment liquid, which is then collected in an aseptic receiving tank. Specifically, the graded filtration process involves: first, pre-filtration through a 0.45μm pore size filter element, followed by sequential passing through sterilization-grade filter elements with 0.22μm and 0.1μm pore sizes.

[0049] S7. Aseptic filling and finished product handling: The refined replenishing solution is filled into pre-sterilized light-proof containers under aseptic conditions; the atmosphere is immediately replaced after filling, and the container is sealed and stored in the dark under refrigeration conditions. The light-proof container is a pre-sterilized brown type III glass bottle; the gas used for atmosphere replacement is nitrogen, with a headspace oxygen content of less than 2.0% v / v; and the refrigeration condition is 5°C.

[0050] Example 3 This application provides a compound polypeptide growth-promoting supplement for livestock and poultry, comprising the following components by weight: 1.0 part recombinant human epidermal growth factor, 1.2 parts trefoil peptide, 0.8 parts insulin-like growth factor, 30 parts vitamin A palmitate, 220 parts vitamin B complex, 340 parts magnesium L-ascorbic acid-2-phosphate, 120 parts D-α-tocopherol succinate, 480 parts recombinant collagen peptide, 120 parts marine-derived acidic polysaccharide, 460 parts compound carrier, 180 parts pH adjuster, and 12 parts stabilizer. The vitamin B complex contains thiamine hydrochloride, sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate in a mass ratio of 1.2:1.4:1.2:2.2:1.7. The composite carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose and low molecular weight sodium hyaluronate in a mass ratio of 8:5:2. Among them, the trifolin peptide is TFF3, the insulin-like growth factor is IGF-2, the pH adjuster is a mixture of sodium hydroxide and citric acid with a mass ratio of 1:3, the stabilizer is a mixture of sorbitol and ascorbyl palmitate with a mass ratio of 20:1, and the marine-derived acidic polysaccharide is fucoidan sulfate oligosaccharide.

[0051] The preparation method of the above-mentioned compound polypeptide growth-promoting rehydration solution for livestock and poultry includes the following steps: S1. Construction and activation of the structured carrier: Hydroxypropyl-β-cyclodextrin and trehalose were dissolved in phosphate buffer to prepare solution A; low molecular weight sodium hyaluronate was dissolved in the same buffer to prepare solution B; under continuous stirring and low temperature conditions, solution B was slowly added dropwise to solution A, and stirring was continued after the addition was completed to obtain the structured composite carrier solution. The phosphate buffer solution has a pH of 7.2; the concentration of solution A is 20% w / v and the concentration of solution B is 5% w / v; the low temperature condition is 8°C; the stirring speed is 300 rpm and the stirring time is 60 min.

[0052] S2. Sequential embedding and stabilization of peptide components: Recombinant collagen peptides are dissolved in the composite carrier solution prepared in part of step S1 to form solution C; while maintaining the system at low temperature, recombinant human epidermal growth factor is first added to solution C, stirred to dissolve and allowed to stand; then, trefoil peptide and insulin-like growth factor are added sequentially, and each peptide is gently stirred at an appropriate speed to ensure full dispersion, thus obtaining the peptide-embedded composite solution. The concentration of solution C was 12.0% w / v; the low temperature condition was 6.0℃; the standing time was 17 min; the gentle stirring speed was 200 rpm; and the stirring time after each polypeptide was added was 12 min.

[0053] S3. Construction of the microenvironment for the vitamin complex system: Marine-derived acidic polysaccharides were dissolved in deionized water to prepare acidic polysaccharide solution D; vitamin A palmitate and D-α-tocopherol succinate were mixed with acidic polysaccharide solution D and emulsified in a water bath to obtain primary fat-soluble microemulsion E; vitamin B complex and magnesium L-ascorbic acid-2-phosphate were dissolved in the remaining complex carrier solution to obtain aqueous vitamin solution F; under room temperature and nitrogen atmosphere, primary fat-soluble microemulsion E was slowly injected into aqueous vitamin solution F and homogenized and circulated to obtain vitamin complex emulsion G; The concentration of acidic polysaccharide solution D is 6.0% w / v; the temperature of the water bath is 55℃; the emulsification speed is 12000 rpm; the emulsification time is 5 min; the pressure of the homogenization cycle is 120 MPa; and the number of cycles is 4.

[0054] S4. Programmed gradient compounding and phase integration: The peptide-loaded composite solution obtained in step S2 is transferred to a reaction vessel and stirred. The vitamin composite emulsion G obtained in step S3 is pumped into the reaction vessel. After pumping, the temperature is programmed and the integration reaction continues at the temperature after the temperature is increased to obtain the integrated solution. The stirring temperature is 12℃; the stirring speed is 250 rpm; the pumping is completed at a constant speed within 50 minutes; the programmed heating rate is 0.5℃ / min, heating to 26℃; the integration reaction speed is 400 rpm, and the reaction time is 150 minutes.

[0055] S5. Dynamic pH control and terminal stabilization: Add a pH adjuster slowly to the integrated solution obtained in step S4 while stirring to precisely adjust the pH value of the system step by step, and then add a stabilizer. Specifically, the stepwise precise adjustment involves first adjusting the pH to 6.8 and holding it for 15 minutes, then fine-tuning it to 7.0 and holding it for 10 minutes.

[0056] S6. Multi-mode terminal refining and aseptic collection: The liquid processed in step S5 is subjected to graded filtration to obtain refined replenishment liquid, which is then collected in an aseptic receiving tank. Specifically, the graded filtration process involves: first, pre-filtration through a 0.45μm pore size filter element, followed by sequential passing through sterilization-grade filter elements with 0.22μm and 0.1μm pore sizes.

[0057] S7. Aseptic filling and finished product handling: The refined replenishing solution is filled into pre-sterilized light-proof containers under aseptic conditions; the atmosphere is immediately replaced after filling, and the container is sealed and stored in the dark under refrigeration conditions. The light-proof container is a pre-sterilized brown type III glass bottle; the gas used for atmosphere replacement is nitrogen, with a headspace oxygen content of less than 2.0% v / v; and the refrigeration condition is 8°C.

[0058] Comparative Example 1 The only difference between this comparative example and Example 1 is that recombinant human epidermal growth factor is not added; the other components, dosages, and preparation methods are exactly the same.

[0059] Comparative Example 2 The only difference between this comparative example and Example 1 is that the recombinant collagen peptides are replaced with an equal amount of ordinary hydrolyzed plant protein soybean peptides, while the other components and preparation methods are exactly the same.

[0060] Comparative Example 3 The only difference between this comparative example and Example 1 is that the marine-derived acidic polysaccharide is replaced with an equal amount of the common emulsifier Tween-80; the other components and preparation methods are exactly the same.

[0061] Comparative Example 4 The only difference between this comparative example and Example 1 is that the composite carrier is replaced with an equal amount of a single carrier, only hydroxypropyl-β-cyclodextrin is used, and step S1 is adjusted accordingly to directly dissolve hydroxypropyl-β-cyclodextrin.

[0062] Comparative Example 5 The only difference between this comparative example and Example 1 is that the separate pretreatment and program integration steps of S2, S3, and S4 are cancelled. Instead, all raw materials except pH adjuster and stabilizer are added to the composite carrier solution at one time, stirred at 25°C and 300 rpm for 60 minutes to mix, and then steps S5-S7 are performed.

[0063] Comparative Example 6 This comparative example uses a commercially available liquid livestock and poultry nutritional supplement whose main components are multiple vitamins and amino acids, but does not contain recombinant polypeptide growth factors, recombinant collagen peptides, or marine-derived acidic polysaccharides.

[0064] Test Item 1: Determination of Daily Weight Gain and Feed Conversion Efficiency in Weaned Piglets The testing standards refer to GB / T34748-2017 "Technical Specification for Determination of Piglet Production Performance"; One hundred and eighty healthy weaned piglets aged 21 days, with a uniform weight of 6.0 ± 0.5 kg, were randomly divided into nine groups of 20 piglets each (half male and half female), corresponding to Examples 1-3 and Comparative Examples 1-6, respectively. All piglets were raised under identical environmental conditions: temperature controlled at 24-26℃, humidity maintained at 55%-65%, and good ventilation. The rearing period was 28 days. Comparative Example 6 served as the control group, receiving a commercially available liquid livestock and poultry nutritional supplement at the recommended dosage. Examples 1-3 and Comparative Examples 1-5 received the same supplemental fluids via drinking water at the same effective ingredient concentration. All groups had free access to feed and water. The total feed intake of each group was recorded daily. At the beginning and end of the rearing period, the piglets were weighed on an empty stomach. The average daily weight gain and feed conversion efficiency (FCE) of each group were calculated. FCE = Total feed intake / Total weight gain.

[0065] Test Item 2: Determination of intestinal health and diarrhea rate in weaned piglets The testing standards refer to NY / T2852-2015 "Technical Specifications for Evaluation of Livestock and Poultry Intestinal Health"; Using the same grouping settings, feeding conditions, and 28-day feeding cycle as Test 1, the incidence of diarrhea in each group of piglets was observed and recorded daily. The criteria for diarrhea were loose or watery feces that were not formed. The diarrhea rate for each group was calculated as follows: Diarrhea rate = Total number of days with diarrhea / (Total number of piglets in the group × Number of feeding days) × 100%. After the feeding cycle, 6 piglets were randomly selected from each group for slaughter. The middle segment of the duodenum and jejunum was quickly separated into 2 cm sections. After rinsing with physiological saline, the sections were fixed in 4% paraformaldehyde solution. After routine paraffin embedding, sectioning, and HE staining, the height of the intestinal mucosa villi and the depth of the crypts were observed under an optical microscope, and the villi height / crypt depth ratio was calculated.

[0066] Test Item 3: Determination of Product Storage Stability and Retention Rate of Active Ingredients The testing standards refer to the determination method of active ingredients in GB / T28643-2012 "Feed Additives - Polypeptides" and GB / T18823-2019 "Permissible Error in Judgment of Feed Test Results"; The replenishment samples from Examples 1-3 and Comparative Examples 1-5 were stored at 2-8°C for 6 months. The sample from Comparative Example 6 was stored according to the recommended conditions in its instructions. Samples were taken and tested at 0 days, 3 months, and 6 months of storage. The contents of recombinant human epidermal growth factor, trefoil peptide TFF2, and insulin-like growth factor IGF-1 were determined by high performance liquid chromatography (HPLC). The retention rate of each active ingredient was calculated as follows: retention rate = (content after storage / initial content) × 100%. The pH value of the samples was measured using a precision pH meter, and the pH value change was recorded.

[0067] The results of the daily weight gain and feed conversion efficiency of weaned piglets are shown in Table 1.

[0068] Table 1:

[0069] The results of the intestinal health and diarrhea rate of weaned piglets are shown in Table 2.

[0070] Table 2:

[0071] The results of the product storage stability and active ingredient retention rate tests are shown in Table 3.

[0072] Table 3:

[0073] Combining Examples 1-3 and Comparative Example 1 with Tables 1 and 2, it can be seen that the deletion of the recombinant human epidermal growth factor (rGF) component leads to a slowdown in the growth rate and a decrease in the integrity of the intestinal structure in weaned piglets. The absence of this growth factor disrupts the synergistic effect between the peptide components in the formulation, preventing them from effectively promoting the proliferation and repair of intestinal epithelial cells, thereby affecting the absorption efficiency of nutrients, manifested as reduced daily weight gain and a decreased villous-crypt structure ratio. This result demonstrates that recombinant human epidermal growth factor, as one of the core active ingredients, plays an irreplaceable role in promoting growth and maintaining healthy intestinal function in this compound system.

[0074] Based on Examples 1-3 and Comparative Example 2, and referring to Tables 1, 2, and 3, it can be seen that replacing recombinant collagen peptides with ordinary hydrolyzed plant protein led to a decline in both the animal performance and storage stability of the product. The specific amino acid sequence and molecular structure of recombinant collagen peptides can specifically interact with peptide growth factors, forming a protective microenvironment. Replacing them with structurally different plant proteins weakens this protective effect, making the active peptides more prone to denaturation or aggregation during preparation and storage, ultimately affecting the product's growth-promoting effect and intestinal health maintenance function. Simultaneously, the retention rate of active ingredients is also significantly reduced.

[0075] Based on Examples 1-3 and Comparative Example 3, and referring to Tables 1, 2, and 3, it can be seen that replacing the marine-derived acidic polysaccharide alginate oligosaccharide with the common chemical emulsifier Tween-80 negatively impacted the overall performance of the product. Marine-derived acidic polysaccharides not only possess emulsifying capabilities, but their unique oligosaccharide structure and surface charge also allow them to form stable complex networks with other components in the system through intermolecular forces. In contrast, Tween-80 only provides a single emulsifying function and cannot construct the same stable microenvironment. Consequently, the dispersion stability of fat-soluble vitamins, the protection of peptide activity, and the final animal application effects are all inferior to those of the examples using marine-derived acidic polysaccharides.

[0076] Based on Examples 1-3 and Comparative Example 4, and referring to Tables 1, 2, and 3, it can be seen that replacing the composite carrier composed of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate with a single hydroxypropyl-β-cyclodextrin carrier significantly reduced the stability and efficacy of the product. The components in the composite carrier work synergistically, providing different functions such as molecular inclusion, cryogenic protection, and solution rheological stability, collectively forming a multi-layered stability system. A single carrier cannot achieve this synergistic protection, leading to accelerated degradation of the active ingredient during storage, decreased physical stability of the product, and directly weakening its growth-promoting and gut health-improving effects in animals.

[0077] As can be seen from Examples 1-3 and Comparative Example 5, and from Tables 1, 2, and 3, eliminating the pretreatment and programmed integration steps and simply mixing all raw materials severely compromises the quality of the final product. Stepwise processing allows for separate optimization of pretreatment conditions for shear-sensitive peptide systems and vitamin systems requiring homogenization and emulsification, while programmed integration allows for gentle and controllable interactions between different systems.

[0078] As can be seen from Examples 1-3 and Comparative Example 6, and in conjunction with Tables 1 and 2, compared with commercially available common nutritional supplements that do not contain the core components of the present invention, such as the specific polypeptide growth factors, recombinant collagen peptides, and marine-derived acidic polysaccharides, the embodiments of this application demonstrate comprehensive advantages in promoting the growth of weaned piglets and improving intestinal health.

[0079] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A compound polypeptide growth-promoting fluid for livestock and poultry, characterized in that: By weight, it comprises the following components: 0.8-1 parts recombinant human epidermal growth factor, 1-1.2 parts trefoil peptide, 0.6-0.8 parts insulin-like growth factor, 20-30 parts vitamin A palmitate, 200-220 parts vitamin B complex, 300-340 parts magnesium L-ascorbic acid-2-phosphate, 80-120 parts D-α-tocopherol succinate, 400-480 parts recombinant collagen peptide, 100-120 parts marine-derived acidic polysaccharide, 380-460 parts complex carrier, 120-180 parts pH adjuster, and 10-12 parts stabilizer.

2. The compound polypeptide growth-promoting fluid for livestock and poultry according to claim 1, characterized in that: The vitamin B complex comprises thiamine hydrochloride, sodium riboflavin-5'-phosphate, pyridoxine hydrochloride, nicotinamide, and D-calcium pantothenate in a mass ratio of (0.8-1.2):(1.0-1.4):(0.8-1.2):(1.8-2.2):(1.3-1.7); the complex carrier is composed of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate, wherein the mass ratio of hydroxypropyl-β-cyclodextrin, trehalose, and low molecular weight sodium hyaluronate is 5:3:1 to 8:5:

2.

3. The compound polypeptide growth-promoting fluid for livestock and poultry according to claim 1, characterized in that: The trifoliate peptide is selected from at least one of TFF1, TFF2, or TFF3; the insulin-like growth factor is selected from IGF-1 or IGF-2; the pH adjuster is selected from at least one of hydrochloric acid, sodium hydroxide, citric acid, sodium citrate, sodium dihydrogen phosphate, or disodium hydrogen phosphate; the stabilizer is selected from at least one of trehalose, L-methionine, sorbitol, mannitol, or ascorbyl palmitate; and the marine-derived acidic polysaccharide is selected from at least one of alginate oligosaccharide, carrageenan oligosaccharide, or fucoidan sulfate oligosaccharide.

4. A method for preparing a compound polypeptide growth-promoting rehydration solution for livestock and poultry, characterized in that, The compound polypeptide growth-promoting supplement for livestock and poultry as described in any one of claims 1-3 comprises the following steps: S1. Construction and activation of the structured carrier: Hydroxypropyl-β-cyclodextrin and trehalose were dissolved in phosphate buffer to prepare solution A; low molecular weight sodium hyaluronate was dissolved in the same buffer to prepare solution B; under continuous stirring and low temperature conditions, solution B was slowly added dropwise to solution A, and stirring was continued after the addition was completed to obtain the structured composite carrier solution. S2. Sequential embedding and stabilization of peptide components: Recombinant collagen peptides are dissolved in the composite carrier solution prepared in part of step S1 to form solution C; while maintaining the system at low temperature, recombinant human epidermal growth factor is first added to solution C, stirred to dissolve and allowed to stand; then, trefoil peptide and insulin-like growth factor are added sequentially, and each peptide is gently stirred at an appropriate speed to ensure full dispersion, thus obtaining the peptide-embedded composite solution. S3. Construction of the microenvironment for the vitamin complex system: Marine-derived acidic polysaccharides were dissolved in deionized water to prepare acidic polysaccharide solution D; vitamin A palmitate and D-α-tocopherol succinate were mixed with acidic polysaccharide solution D and emulsified in a water bath to obtain primary fat-soluble microemulsion E; vitamin B complex and magnesium L-ascorbic acid-2-phosphate were dissolved in the remaining complex carrier solution to obtain aqueous vitamin solution F; under room temperature and nitrogen atmosphere, primary fat-soluble microemulsion E was slowly injected into aqueous vitamin solution F and homogenized and circulated to obtain vitamin complex emulsion G; S4. Programmed gradient compounding and phase integration: The peptide-loaded composite solution obtained in step S2 is transferred to a reaction vessel and stirred. The vitamin composite emulsion G obtained in step S3 is pumped into the reaction vessel. After pumping, the temperature is programmed and the integration reaction continues at the temperature after the temperature is increased to obtain the integrated solution. S5. Dynamic pH control and terminal stabilization: Add a pH adjuster slowly to the integrated solution obtained in step S4 while stirring to precisely adjust the pH value of the system step by step, and then add a stabilizer. S6. Multi-mode terminal refining and aseptic collection: The liquid processed in step S5 is subjected to graded filtration to obtain refined replenishment liquid, which is then collected in an aseptic receiving tank. S7. Aseptic filling and finished product handling: The refined replenishing solution is filled into pre-sterilized light-proof containers under aseptic conditions; the atmosphere is immediately replaced after filling, and the container is sealed and stored in the dark under refrigeration conditions.

5. The method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry according to claim 4, characterized in that: In step S1, the pH value of the phosphate buffer solution is 6.8 to 7.2; the concentration of solution A is 10-20% w / v, and the concentration of solution B is 2-5% w / v; the low temperature condition is 4-8℃; the stirring speed is 100-300 rpm, and the stirring time is 30-60 min.

6. The method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry according to claim 4, characterized in that: In step S2, the concentration of solution C is 8.0-12.0% w / v; the low temperature condition is 2.0-6.0℃; the standing time is 15-17 min; the gentle stirring speed is 100-200 rpm; and the stirring time after each polypeptide is added is 10-12 min.

7. The method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry according to claim 4, characterized in that: In step S3, the concentration of the acidic polysaccharide solution D is 3.0% w / v to 6.0% w / v; the temperature of the water bath is 45-55℃; the emulsification speed is 8000-12000 rpm; the emulsification time is 3-5 min; the pressure of the homogenization cycle is 80-120 MPa; and the number of cycles is 2-4.

8. The method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry according to claim 4, characterized in that: In step S4, the stirring temperature is 8-12℃; the stirring speed is 150-250 rpm; the pumping is completed at a constant speed within 30-50 minutes; the programmed temperature rise rate is 0.3-0.5℃ / min, and the temperature rises to 22-26℃; the rotation speed of the integration reaction is 300-400 rpm, and the reaction time is 90-150 minutes.

9. The method for preparing a compound polypeptide growth-promoting supplement for livestock and poultry according to claim 4, characterized in that: In step S5, the stepwise precise adjustment specifically involves: first adjusting the pH to 6.2 to 6.8 and maintaining it for 15 minutes, then fine-tuning it to 6.5 to 7.0 and maintaining it for 5-10 minutes; in step S6, the graded filtration specifically involves: first pre-filtration through a 0.45μm pore size filter element, then sequentially passing through sterile filter elements with 0.22μm and 0.1μm pore sizes; in step S7, the light-proof container is a pre-sterilized brown type III glass bottle; the gas used for atmosphere replacement is nitrogen, wherein the headspace oxygen content is less than 2.0% v / v; the refrigeration conditions are 2-8℃.

10. An application of a compound polypeptide growth-promoting fluid for livestock and poultry, characterized in that, This is a compound polypeptide growth supplement for livestock and poultry as described in any one of claims 1-3, used to prepare feed additives or drinking water additives that promote livestock and poultry growth, maintain intestinal health, and improve feed utilization.

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