High-nutrition egg feed rich in astaxanthin for egg gamecocks and preparation process of high-nutrition egg feed

Abstract

The invention relates to the field of animal feeds, and discloses an astaxanthin-rich high-nutrition egg feed for egg gamecocks and a preparation process of the astaxanthin-rich high-nutrition egg feed for egg gamecocks, and the feed comprises the following components in percentage by weight: 1.0-3.0% of enzymolysis yeast or algae source astaxanthin biomass; the invention discloses an astaxanthin product coated with a liposome-chitosan composite micro-capsule. Functional additives such as L-arginine, creatine monohydrate and the like; and a basal feed matrix. The preparation process comprises the following steps: preparing an astaxanthin product coated with enzymolysis astaxanthin biomass and lipidosome-chitosan composite micro-capsules; mixing the basic feed components with the enzymolysis astaxanthin biomass and the like, and then performing low-temperature granulation; and then performing vacuum dynamic coating on the obtained basic particles, and spraying liquid containing the microcapsule astaxanthin and other thermosensitive active ingredients. The stability and bioavailability of astaxanthin and other active ingredients in the feed processing and storage process and the animal digestion process are remarkably improved, deposition of astaxanthin in egg yolk is effectively promoted, and the quality of egg products is improved.

Description

Technical Field

[0001] The invention relates to the technical field of animal feed, in particular to a high-nutrition egg feed for fighting chickens rich in astaxanthin and a preparation process thereof. Background Art

[0002] Astaxanthin, a potent natural antioxidant, has garnered widespread attention in aquaculture and poultry farming. It not only imparts a desirable color to egg yolks and poultry meat, but also enhances animals' immune function, stress resistance, and reproductive performance. Currently, the addition of astaxanthin to egg-laying poultry feed to improve yolk color and enhance the nutritional value of eggs is a common practice. However, existing technologies still have some shortcomings in terms of the efficient utilization of astaxanthin and the synergistic improvement of the overall nutritional efficacy of feed.

[0003] On the one hand, astaxanthin is chemically active and sensitive to environmental factors such as light, heat, and oxygen. It is easily degraded and inactivated under the high temperature, high humidity, and high pressure conditions of conventional feed processing, resulting in its actual effective content in the final feed product being far lower than the added amount, and its bioavailability is low. Although studies have attempted to use coating technology to improve the stability of astaxanthin, a single coating method often cannot balance protection during processing and effective release and absorption in the digestive tract. In addition, astaxanthin in natural astaxanthin sources (such as algae powder and yeast) is often encapsulated by structures such as cell walls, and its release and absorption efficiency is limited when added directly.

[0004] On the other hand, for poultry species with specific uses, such as egg-laying fighting chickens with their unique physiological needs, their feed must not only improve egg quality but also take into account their high energy, robust physique, and good reproductive performance. Existing feed formulas do not fully consider the comprehensive nutritional regulation of these species, lacking systematic optimization of the synergistic effects between astaxanthin and other functional nutrients (such as specific amino acids, vitamins, and probiotics that promote energy metabolism, enhance immunity, and improve reproductive performance). Therefore, the development of a highly nutritious feed that can ensure the efficient and stable utilization of astaxanthin and synergistically improve the overall physiological functions and egg quality of egg-laying fighting chickens, as well as a supporting gentle and efficient preparation process, has important practical application value. Summary of the Invention

[0005] The present invention aims to overcome the shortcomings of the existing egg-laying poultry feed in terms of astaxanthin utilization, active ingredient stability, and synergistic improvement of the physiological functions of specific poultry species (such as fighting chickens), and to provide a new type of astaxanthin-rich high-nutrition egg feed for fighting chickens and its efficient and gentle preparation process.

[0006] To achieve the above objectives, the present invention is implemented through the following technical solutions: The first aspect of the present invention provides a high-nutrition egg feed for fighting chickens rich in astaxanthin, which comprises, by weight percentage: (a) 1.0-3.0% enzymatically hydrolyzed yeast or algae-derived astaxanthin biomass; (b) an astaxanthin product coated with 0.05-0.15% liposome-chitosan composite microcapsules, wherein the astaxanthin encapsulation content of the astaxanthin product is 8-12%; (c) at least one functional additive selected from the following: 0.5-1.5% L-arginine; 0.3-1.0% creatine monohydrate; 0.06-0.15% L-ascorbic acid-2-polyphosphate; 0.02-0.1% of a composite probiotic preparation comprising at least one of Bacillus coagulans, Enterococcus faecalis, or Lactobacillus plantarum; (d) A basic feed matrix comprising a protein source, an energy source, vitamins and minerals.

[0007] The feed described in this invention combines two astaxanthin sources with different forms and delivery mechanisms: enzymatically treated natural astaxanthin biomass and highly purified astaxanthin encapsulated with liposome-chitosan composite microcapsules. The goal is to improve the stability and bioavailability of astaxanthin during feed processing, storage, and animal digestion. Enzymatic treatment facilitates the release of natural astaxanthin from cellular structures, while the liposomes mimic the cell membrane structure, promoting intestinal cell affinity and absorption of astaxanthin. The chitosan shell provides physical protection for astaxanthin and potentially enables sustained release in specific areas of the digestive tract. This dual astaxanthin delivery system works synergistically, facilitating its effective absorption in fighting cocks and efficient deposition in egg yolks.

[0008] Furthermore, the enzymatic yeast or algal astaxanthin biomass (a) is derived from fermented Phaffia rhodozyma or Haematococcus pluvialis, both of which are recognized as high-quality natural astaxanthin sources. The liposome-chitosan composite microcapsule-encapsulated astaxanthin product (b) is prepared using high-purity astaxanthin with a purity of ≥95%, ensuring the quality and efficacy of the encapsulated core material.

[0009] In addition, specific functional additives added to the feed, such as L-arginine and creatine monohydrate, are designed to support muscle energy metabolism and functional maintenance in fighting cocks. L-ascorbic acid-2-polyphosphate, a stable source of vitamin C, synergizes with astaxanthin to exert antioxidant effects and enhance the body's resistance to stress. A complex probiotic formulation helps maintain the balance of the fighting cocks' intestinal microecology, improve digestion and absorption, and support immune health. These ingredients are scientifically combined to comprehensively enhance the fighting cocks' physiological functions and the nutritional quality of their eggs.

[0010] In some embodiments, the feed may also contain 0.05-0.2% L-carnitine to promote fatty acid oxidation for energy supply; 0.03-0.06% DL-α-tocopheryl acetate, as an important fat-soluble antioxidant, synergistically acting with astaxanthin and vitamin C; 0.015-0.03% organic selenium to enhance the activity of the antioxidant enzyme system; biotin added in an amount of 0.2-0.5 mg / kg based on feed, and 0.1-0.3% methionine zinc complex to support feather and skin health; 0.5-1.5% small molecule collagen peptides, which may help tissue repair and joint health.

[0011] For breeding fighting cocks, the feed can further include 1.0-3.0% fish oil rich in EPA and DHA to improve reproductive performance and egg quality; folic acid added in an amount of 2.0-5.0 mg / kg based on feed, and vitamin B12 added in an amount of 0.02-0.05 mg / kg based on feed to meet the special nutritional needs during the breeding period.

[0012] A second aspect of the present invention provides a method for preparing the above-mentioned high-nutrition egg feed for fighting chickens rich in astaxanthin, comprising the following steps: (i) preparing enzymatic yeast or algae-derived astaxanthin biomass; (ii) preparing an astaxanthin product coated with liposome-chitosan composite microcapsules; (iii) uniformly mixing the basic feed components with the enzymatic yeast or algal astaxanthin biomass obtained in step (i) and any other heat-resistant additives, and performing low-temperature granulation to form basic granules, wherein the granule temperature does not exceed 65° C. during the granulation process; (iv) vacuum dynamically coating the base particles obtained in step (iii) by spraying a liquid mixture comprising the astaxanthin product encapsulated by the liposome-chitosan composite microcapsules obtained in step (ii) and other thermosensitive active ingredients under vacuum conditions of -0.05 to -0.08 MPa relative pressure.

[0013] The preparation method provided by the present invention maximizes the bioactivity of astaxanthin and other heat-sensitive active ingredients through step-by-step processing and specific process conditions. In step (i), the natural astaxanthin biomass is subjected to enzymatic pretreatment to disrupt the cell wall structure, thereby increasing the release rate and subsequent availability of astaxanthin. The liposome-chitosan composite microcapsules prepared in step (ii) provide a dual protection and sustained-release system for high-purity astaxanthin, enabling it to better resist the adverse effects of the digestive tract environment and potentially promoting its absorption.

[0014] The low-temperature granulation process in step (iii), for example, cold pressing granulation, controls the temperature of the material before entering the granulator die to no more than 40°C, and the granule discharge temperature does not exceed 60-65°C. This significantly reduces the damage caused by high temperature during the granulation process to the activity of the enzymatically hydrolyzed astaxanthin biomass and other added semi-heat-stable ingredients.

[0015] The key step (iv) uses vacuum dynamic coating technology to evenly spray and infiltrate the more sensitive liposome-chitosan composite microencapsulated astaxanthin prepared in step (ii), as well as other heat-sensitive vitamins, probiotics and other active ingredients, onto the pre-made low-temperature base particles under vacuum conditions. The vacuum environment helps the liquid ingredients penetrate into the pores of the particles, reducing the oxidation of active substances during the spraying process, while avoiding the damage of high temperature to these high-value sensitive ingredients. The amount of liquid mixture sprayed is controlled to 2-5% of the total weight of the base particles to ensure effective coating without affecting the physical properties of the particles. The entire vacuum dynamic coating process is carried out at a temperature of 20-35°C, further ensuring the stability of the active ingredients.

[0016] In the specific implementation of step (i), the yeast or algae-derived astaxanthin biomass can be mixed with a buffer solution with a pH of 4.5-5.5, and a complex enzyme preparation accounting for 0.5-2.0% of the dry weight of the biomass is added, and hydrolyzed at 45-55° C. for 2-6 hours, followed by enzyme inactivation and drying.

[0017] In the specific implementation of step (ii), high-purity astaxanthin, phosphatidylcholine and cholesterol can be dissolved to form a lipid membrane, which is hydrated to form a crude liposome suspension, and then ultrasonically treated with a power of 100-300 W for 5-20 minutes to form nano-astaxanthin liposomes; the nano-astaxanthin liposomes are then added dropwise to a 0.1-0.5% (w / v) chitosan solution with a pH of 4.0-5.5 for coating, and the microcapsules are collected and dried.

[0018] The above-mentioned feed composition and its preparation method can provide a high-quality feed for egg-laying fighting chickens with comprehensive nutrition, good astaxanthin enrichment effect and stable active ingredients, thereby improving the health of fighting chickens, enhancing their production performance and egg quality.

[0019] In summary, the present invention includes at least one of the following beneficial technical effects: 1. This invention utilizes a dual delivery system consisting of enzymatically hydrolyzed yeast / algae-derived astaxanthin biomass and liposome-chitosan composite microcapsules to significantly improve the stability and bioavailability of astaxanthin during feed processing, storage, and animal digestion. Enzymatic hydrolysis promotes the release of natural astaxanthin, while the liposome-chitosan microcapsules provide physical protection and targeted sustained-release of the high-purity astaxanthin, ensuring that more active astaxanthin is effectively absorbed by the fighting cocks and deposited in their egg yolks.

[0020] 2. This invention utilizes a scientific combination of functional additives, including L-arginine, creatine, stabilized vitamin C (L-ascorbic acid 2-polyphosphate), and a complex probiotic, to specifically improve muscle energy metabolism, enhance stress resistance, regulate intestinal microecological balance, and support immune health in fighting cocks. These components, combined with the highly absorbable astaxanthin, contribute to a comprehensive improvement in the cocks' physiological functions, athletic performance, and overall health.

[0021] 3. The present invention's combined low-temperature granulation and vacuum dynamic post-spraying process effectively protects the bioactivity of astaxanthin and other heat- and oxygen-sensitive active ingredients (such as vitamins and probiotics). Low-temperature granulation prevents high-temperature damage to less heat-resistant components, while vacuum dynamic post-spraying allows the highly active and sensitive core ingredients to be evenly coated on the surface and interior of the granules under mild, anaerobic or low-oxygen conditions, minimizing their loss during processing.

[0022] 4. Targeted at the needs of breeding gamecocks, this invention specifically incorporates fertility optimizers such as EPA / DHA-rich fish oil, folic acid, and vitamin B12, combined with highly efficient astaxanthin, to improve the reproductive performance of breeding chickens. These nutrients have a positive effect on enhancing sperm and egg quality, increasing the fertilization and hatchability of breeding eggs, and promoting the healthy early development of chicks, thereby improving overall breeding efficiency.

[0023] 5. The feed composition and preparation method provided by the present invention not only focuses on enriching astaxanthin in eggs, but also on improving the health and production performance of the fighting cocks themselves. By optimizing the astaxanthin delivery method and the ratio of synergistic nutrients, the fighting cocks are made stronger and more disease-resistant. The resulting eggs not only have a high astaxanthin content and ideal yolk color, but also improve the overall nutritional value and sensory quality of the eggs. DETAILED DESCRIPTION

[0024] The following examples are used to further illustrate the present invention, but are not intended to limit the scope of the present invention. Those skilled in the art may make various non-essential improvements and adjustments to the present invention based on the contents of this specification, and these improvements and adjustments still fall within the scope of protection of the present invention.

[0025] Unless otherwise specified, all raw materials used in the examples are commercially available or prepared by conventional methods in the art, and the equipment used is conventional equipment in the art. Unless otherwise specified, percentages and parts are by weight.

[0026] Example A: Preparation of astaxanthin biomass from Haematococcus pluvialis by enzymatic hydrolysis 1. Raw material formula: Haematococcus pluvialis dried cell powder (natural astaxanthin content 0.45% w / w): 1000 g; Citric acid-sodium hydrogen phosphate buffer (0.1 M, pH 5.0): 7000 ml; Complex enzyme preparation (including cellulase activity ≥30,000 U / g, β-glucanase activity ≥20,000 U / g): 15 g.

[0027] 2. Preparation steps: 1000 g of dried Haematococcus pluvialis cell powder was added to 7000 ml of pH 5.0 citric acid-sodium hydrogen phosphate buffer and stirred to form a uniform suspension. 15 g of a complex enzyme preparation was then added, and the mixture was stirred at 150 rpm in a 50°C water bath for 4 hours. After the reaction, the mixture was heated to 85°C for 15 minutes to inactivate the enzyme preparation. The inactivated mixture was spray-dried with the inlet temperature controlled at 160-180°C and the outlet temperature controlled at 75-85°C to obtain a powdered enzymatically hydrolyzed Haematococcus pluvialis astaxanthin biomass, which was then collected for later use. The astaxanthin content was determined to be approximately 0.43% w / w.

[0028] Example B: Preparation of astaxanthin liposome-chitosan composite microcapsules 1. Raw material formula: Astaxanthin crystalline powder (purity ≥96%): 2.5 g; Soy lecithin (phosphatidylcholine content ≥ 70%): 30g; Cholesterol (purity ≥98%): 7.5 g; Mixed solvent of anhydrous ethanol and chloroform (volume ratio 1:2): 200 ml; Phosphate buffered saline (PBS, 0.01 M, pH 7.2): 500 ml; Chitosan (degree of deacetylation 88%, average molecular weight 120 kDa): 5 g; Glacial acetic acid: 5 ml; Deionized water: appropriate amount.

[0029] 2. Preparation steps: a. Preparation of Astaxanthin Liposomes: 2.5 g of crystalline astaxanthin powder, 30 g of soy lecithin, and 7.5 g of cholesterol were dissolved in 200 ml of a mixture of anhydrous ethanol and chloroform. The organic solvent was removed using a rotary evaporator under reduced pressure (approximately -0.08 MPa) at 50°C until a uniform lipid film formed on the inner wall of the flask. 500 ml of pH 7.2 phosphate buffer was added to the flask containing the lipid film, and the mixture was hydrated in a 37°C water bath for 1.5 hours with intermittent shaking to form a crude liposome suspension. This crude liposome suspension was transferred to an ice bath and sonicated using a probe sonicator (power setting 200 W) in a 3-second on, 2-second off mode for a total of 15 minutes to obtain a nanosized astaxanthin liposome suspension.

[0030] b. Chitosan Coating: Weigh 5 g of chitosan and slowly add it to 495 mL of deionized water containing 5 mL of glacial acetic acid. Stir and dissolve to prepare a 1.0% (w / v) chitosan solution. Adjust the pH to 5.0 with 0.1 M NaOH. Slowly and uniformly add the nano-astaxanthin liposome suspension prepared in step a to the chitosan solution at room temperature (25°C) and continuous stirring at 300 rpm. The volume ratio of liposome suspension to chitosan solution is 1:2. After the addition is complete, continue stirring at room temperature for 60 minutes. After the reaction is complete, centrifuge the mixture at 8000 × g for 20 minutes to collect the precipitate. Wash the precipitate twice with deionized water, repeating the same centrifugation procedure after each wash. The final collected precipitate was vacuum freeze-dried for 24 hours to obtain powdered astaxanthin liposome-chitosan composite microcapsules, and the astaxanthin encapsulation content was determined to be approximately 9.8% (w / w), which was collected for later use.

[0031] Example 1: Preparation of high-nutrition egg feed for fighting chickens rich in astaxanthin 1. Feed formula: The weight percentages of the components of the astaxanthin-rich high-nutrition egg feed for fighting chickens of this embodiment are as follows: Enzymatically hydrolyzed astaxanthin biomass from Haematococcus pluvialis (from Example A): 2.0%; Liposome-chitosan composite microcapsule-encapsulated astaxanthin product (from Example B): 0.1%; L-arginine: 1.0%; Creatine monohydrate: 0.5%; L-ascorbic acid-2-polyphosphate (vitamin C content 35%): 0.1%; Composite probiotic preparation (Bacillus coagulans, Enterococcus faecalis, Lactobacillus plantarum, total viable count 2.0×10 10CFU / g): 0.05%; DL-α-tocopheryl acetate (vitamin E potency 500 IU / g): 0.04%; Corn (extrusion): 50.0%; Soybean meal (expanded, crude protein content 46%): 25.0%; Wheat flour: 8.11%; Fish meal (imported white fish meal, crude protein content 65%): 5.0%; Plasma protein powder (porcine source): 1.5%; Soybean oil (grade 1): 3.0%; Calcium hydrogen phosphate (feed grade): 1.5%; Stone powder (calcium carbonate content 38%): 1.0%; Sodium chloride (feed grade): 0.3%; Vitamin and trace element premix (meeting the nutritional needs of fighting chickens during the laying period): 0.8%; The liposome-chitosan composite microcapsule-encapsulated astaxanthin product, L-ascorbic acid-2-polyphosphate, compound probiotic preparation, and DL-α-tocopheryl acetate are added via a subsequent vacuum dynamic spraying process. The remaining components are used to prepare the base particles.

[0032] 2. Preparation process: a. Premixing: Accurately weigh corn, soybean meal, wheat bran, fish meal, plasma protein powder, soybean oil, calcium hydrogen phosphate, stone powder, sodium chloride, vitamin and trace element premix, L-arginine, creatine monohydrate, and enzymatically hydrolyzed Haematococcus pluvialis-derived astaxanthin biomass according to the above formula proportions, place them in a horizontal ribbon mixer, and mix for 15 minutes to ensure uniform mixing and keep the material variation coefficient within 5%.

[0033] b. Low-temperature granulation: Transfer the premixed material to the cold-pressed pelletizer. Maintain the material temperature at room temperature (approximately 25°C) before entering the pelletizer's die. Use a 3.0 mm ring die aperture, a 10:1 compression ratio of effective die length to aperture, and a 0.2 mm gap between the die roller and the die. Start the pelletizer, adjust the feed rate, and monitor the pellet discharge temperature to ensure it does not exceed 60°C. Immediately cool the pellets using ambient air in a countercurrent cooler until the pellet temperature drops to within 3°C of the ambient temperature.

[0034] c. Vacuum dynamic spraying: i. Preparation of the spray liquid: Pre-moisten the formulated amount of the liposome-chitosan composite microcapsule-encapsulated astaxanthin product with a small amount of purified water, then disperse it in an appropriate amount of purified water to form a uniform suspension. Dissolve the formulated amount of L-ascorbic acid-2-polyphosphate in an appropriate amount of purified water. Mix the formulated amount of DL-α-tocopheryl acetate with a small amount of a food-grade emulsifier, then disperse it in purified water to form an emulsion. Disperse the formulated amount of the composite probiotic preparation in an appropriate amount of purified water to form a suspension. Evenly mix the above four liquids or suspensions to obtain a mixed liquid to be sprayed.

[0035] ii. Spraying: Transfer the cooled base particles prepared in step b to the drum of a vacuum coating machine. Seal the drum and start the vacuum pump to a relative pressure of -0.07 MPa. Rotate the drum at 10 rpm and evenly spray the prepared mixed liquid onto the rotating particles through an atomizing nozzle. The total amount of liquid sprayed should be 3.0% of the total weight of the base particles. The entire spraying process should be carried out at an ambient temperature of 28°C for 20 minutes.

[0036] d. Final Drying: After vacuum spraying, the feed pellets are immediately transferred to a low-temperature fluidized bed dryer. Drying is performed for 60 minutes with the inlet air temperature controlled at 45°C, reducing the moisture content of the finished feed to 11.0% (w / w). The dried feed is cooled, screened, and packaged to produce a highly nutritious astaxanthin-rich egg feed for laying hen game.

[0037] Example 2: Preparation of high-nutrition egg feed for fighting chickens rich in astaxanthin 1. Feed formula: The weight percentages of the components of the astaxanthin-rich high-nutrition egg feed for fighting chickens of this embodiment are as follows: Enzymatically hydrolyzed astaxanthin biomass from Haematococcus pluvialis (from Example A): 1.5%; Liposome-chitosan composite microcapsule-encapsulated astaxanthin product (from Example B): 0.12%; L-arginine: 0.8%; Creatine monohydrate: 0.7%; L-ascorbic acid-2-polyphosphate (vitamin C content 35%): 0.08%; Composite probiotic preparation (Bacillus coagulans, Enterococcus faecalis, Lactobacillus plantarum, total viable count 2.0×10 10 CFU / g): 0.03%; Methionine zinc complex (methionine content 15%, zinc content 15%): 0.2%; Corn (extrusion): 51.57%; Soybean meal (expanded, crude protein content 46%): 24.0%; Wheat flour: 8.0%; Fish meal (imported white fish meal, crude protein content 65%): 4.5%; Plasma protein powder (porcine source): 1.0%; Soybean oil (grade 1): 3.5%; Calcium hydrogen phosphate (feed grade): 1.8%; Stone powder (calcium carbonate content 38%): 1.1%; Sodium chloride (feed grade): 0.3%; Vitamin and trace element premix (meeting the nutritional needs of fighting chickens during the laying period): 0.8%; The liposome-chitosan composite microcapsule-encapsulated astaxanthin product, L-ascorbic acid-2-polyphosphate, and composite probiotic preparation are added via a subsequent vacuum dynamic spraying process. The remaining components are used to prepare the base particles.

[0038] 2. Preparation process: The preparation process is the same as that of Example 1.

[0039] Example 3: Preparation of Astaxanthin-Rich High-Nutrient Egg Feed for Egg-laying Chickens (Containing Fertility Optimizer) 1. Feed formula: The weight percentages of the components of the astaxanthin-rich high-nutrition egg feed for fighting chickens of this embodiment are as follows: Enzymatically hydrolyzed astaxanthin biomass from Haematococcus pluvialis (from Example A): 2.5%; Liposome-chitosan composite microcapsule-encapsulated astaxanthin product (from Example B): 0.08%; L-arginine: 1.2%; Creatine monohydrate: 0.4%; L-ascorbic acid-2-polyphosphate (vitamin C content 35%): 0.12%; Composite probiotic preparation (Bacillus coagulans, Enterococcus faecalis, Lactobacillus plantarum, total viable count 2.0×10 10 CFU / g): 0.06%; DL-α-tocopheryl acetate (vitamin E potency 500 IU / g): 0.05%; EPA- and DHA-rich fish oil (total EPA+DHA content 30%): 2.0%; Folic acid (purity ≥98%, added in the form of 0.1% premix, actual addition amount is 0.3%, providing 3mg / kg feed folic acid): 0.3%; Vitamin B12 (cyanocobalamin, added as a 0.1% premix, the actual addition amount is 0.003%, providing 0.03 mg / kg feed vitamin B12): 0.003%; Corn (extrusion): 47.087%; Soybean meal (expanded, crude protein content 46%): 26.0%; Wheat flour: 7.0%; Fish meal (imported white fish meal, crude protein content 65%): 5.5%; Plasma protein powder (porcine source): 1.2%; Calcium hydrogen phosphate (feed grade): 1.4%; Stone powder (calcium carbonate content 38%): 1.0%; Sodium chloride (feed grade): 0.3%; Vitamin and trace element premix (in line with the nutritional needs of breeding fighting chickens): 0.8%; The liposome-chitosan composite microcapsule-encapsulated astaxanthin product, L-ascorbic acid 2-polyphosphate, a probiotic complex, DL-α-tocopheryl acetate, EPA- and DHA-rich fish oil, a folic acid premix, and a vitamin B12 premix are subsequently added via a vacuum dynamic spraying process. The remaining ingredients (excluding the fish oil, which is added for spraying) are used to prepare the base pellets. If a base oil is added to the base diet, it should be added to the base pellets according to the original formulation.

[0040] 2. Preparation process: The preparation process is the same as in Example 1, except that, in step c1, when preparing the spray liquid, the formulated amounts of EPA- and DHA-rich fish oil, folic acid premix, and vitamin B12 premix are also added to the mixed liquid to be sprayed. The total amount of spray liquid is controlled to be 4.0% of the total weight of the base particles.

[0041] Comparative Example 1: Feed using astaxanthin without special treatment and conventional high-temperature granulation Compared with Example 1, the difference is: 1. Astaxanthin Source: This comparative example did not use the enzymatically hydrolyzed Haematococcus pluvialis-derived astaxanthin biomass prepared in Example A or the liposome-chitosan composite microcapsule-encapsulated astaxanthin product prepared in Example B. Instead, 0.07% of commercially available Haematococcus pluvialis powder (astaxanthin content: 3.5% w / w) was added directly to achieve a theoretical total astaxanthin addition level similar to that in Example 1. This Haematococcus pluvialis powder was premixed with the other basal diet ingredients.

[0042] 2. Preparation Process: This comparative example does not utilize low-temperature granulation or vacuum dynamic spraying. All components (including common Haematococcus pluvialis powder, L-arginine, creatine monohydrate, L-ascorbic acid 2-polyphosphate, compound probiotic preparation, DL-α-tocopheryl acetate, etc.) are premixed and then directly subjected to a conventional high-temperature granulation process. This involves steam conditioning (85°C for 30 seconds) before pelleting in a pelletizer. The pellets are discharged at a temperature of approximately 80-90°C, followed by cooling and drying. The remaining component types and proportions, as well as the final feed form, are essentially the same as in Example 1. The amount of corn in the basal diet is adjusted to account for the weight difference.

[0043] Comparative Example 2: Feed containing specially treated astaxanthin components but using overall high temperature granulation Compared with Example 1, the difference is: 1. Preparation Process: This comparative example utilizes the same types and amounts of enzymatically hydrolyzed Haematococcus pluvialis-derived astaxanthin biomass (from Example A) and liposome-chitosan composite microencapsulated astaxanthin product (from Example B), as well as the same other functional additives, as in Example 1. However, low-temperature granulation and vacuum dynamic spraying are not employed. All components (including the enzymatically hydrolyzed astaxanthin biomass, microencapsulated astaxanthin, and all other heat-sensitive additives such as L-ascorbic acid-2-polyphosphate, the composite probiotic preparation, and DL-α-tocopheryl acetate) are premixed and then directly subjected to a conventional high-temperature granulation process along with the basal diet components. This involves steam conditioning (85°C for 30 seconds) before pelleting in a pelletizer. The pellets are discharged at a temperature of approximately 80-90°C, followed by cooling and drying. The remaining component types and proportions are the same as in Example 1.

[0044] Test Example 1: Experimental description: This test example is intended to evaluate the stability of the key active ingredients astaxanthin and vitamin C in the feed prepared in Example 1 of the present invention and the comparative feed under accelerated storage conditions.

[0045] The astaxanthin-rich high-nutrition egg feed for fighting chickens prepared in Example 1, the feed prepared in Comparative Example 1 using astaxanthin without special treatment and using conventional high-temperature granulation, and the feed prepared in Comparative Example 2 containing a specially treated astaxanthin component but using overall high-temperature granulation were selected as test samples.

[0046] 1. The experimental steps are as follows: Initial Content Determination: Accurately weigh three portions of each of the three freshly prepared feed samples, approximately 10 grams each. For astaxanthin content, extract the samples with an organic solvent (e.g., acetone or a dichloromethane / methanol mixture). After appropriate purification, the extract is analyzed by high-performance liquid chromatography (HPLC) at a specific wavelength (e.g., 470-480 nm). For vitamin C content, extract the samples with a dilute acid solution and determine the active vitamin C content by HPLC (e.g., equipped with a UV detector, detecting L-ascorbic acid-2-polyphosphate at 254 nm or 265 nm and converting to active vitamin C content) or iodine titration. Record the initial average astaxanthin and vitamin C content (mg / kg feed) for each sample.

[0047] 2. Accelerated storage test: Take appropriate amounts of the three feed samples (about 500 g / portion) and place them in breathable packaging bags. Place them in a constant temperature and humidity incubator. Set the storage conditions to 37±1°C and 70±5% relative humidity. Store away from light for 30 days.

[0048] 3. Post-Storage Content Determination: After 30 days of accelerated storage, take three samples from each feed sample and determine the astaxanthin and vitamin C content in each feed sample using the same method as in Step 1. Record the average post-storage astaxanthin and vitamin C content (mg / kg feed) for each sample.

[0049] 4. Evaluation indicators: Calculate the retention rate of astaxanthin and vitamin C in each sample before and after storage.

[0050] Retention rate (%) = (average content after storage / initial average content) × 100%.

[0051] The experimental data are shown in Table 1: Table 1: Retention rate of active ingredients in different feed samples under accelerated storage conditions (%) The data in Table 1 show that the retention rate of astaxanthin in the feed prepared in Example 1 is significantly higher than that in Comparative Example 1 and Comparative Example 2 under accelerated storage conditions. This is mainly due to the use of a dual astaxanthin delivery system and a specific low-temperature preparation process in Example 1. Among them, the enzymatic treatment of natural astaxanthin biomass improves the release of astaxanthin, and the liposome-chitosan composite microcapsule technology provides an effective physical and chemical protection barrier for high-purity astaxanthin, reducing its degradation caused by factors such as oxidation and light during storage. In contrast, Comparative Example 1 directly uses ordinary astaxanthin raw materials and adopts high-temperature granulation, resulting in a large loss of astaxanthin. Even though Comparative Example 2 uses the same specially treated astaxanthin component as Example 1, due to its use of an overall high-temperature granulation process, the retention rate of astaxanthin is much lower than that of Example 1, which further confirms the importance of the low-temperature granulation and vacuum post-spraying process of the present invention in protecting the activity of astaxanthin.

[0052] As a heat-sensitive and easily oxidized active ingredient, vitamin C's stability is also significantly affected by feed processing techniques. In Example 1, a stable vitamin C derivative (L-ascorbic acid-2-polyphosphate) was added to the cooled base granules via low-temperature vacuum dynamic spraying, avoiding the destruction of its activity during the high-temperature granulation process and thus achieving a high retention rate. Comparative Examples 1 and 2, due to the use of high-temperature granulation processes, suffered significant losses during processing and storage, even when using the same or similar vitamin C raw materials. This result highlights the unique advantage of the low-temperature spraying process in the later stages of the present method for protecting heat-sensitive active substances.

[0053] In summary, the present invention, through a preparation method combining specific active ingredient pretreatment technologies (such as enzymatic hydrolysis and microencapsulation of astaxanthin) with optimized low-temperature granulation and vacuum dynamic post-spraying, effectively improves the stability of key nutrients such as astaxanthin and vitamin C in feed during processing and storage. This ensures the effective content of active ingredients in the final feed product, laying a solid foundation for exerting their physiological benefits, improving the health of laying game chickens and the quality of their eggs, and demonstrating the significant beneficial effects of the present invention's technical solution in improving feed quality.

[0054] Test Example 2: Experimental description: This test example aims to evaluate the deposition effect of astaxanthin in the yolk and the yolk color of eggs produced by egg-laying fighting chickens after feeding the feed prepared in Example 1 of the present invention.

[0055] Ninety Thai fighting chickens of 30 weeks old, in good health and with similar egg production rates were randomly divided into three groups, with 30 chickens in each group (three replicates per group and 10 chickens in each replicate).

[0056] The experimental steps are as follows: 1. Experimental groups and feed: Group 1 of Example 1: The game chickens were fed with the high-nutrition egg feed rich in astaxanthin prepared in Example 1.

[0057] Comparative Example 1 group: The rats were fed with the feed prepared in Comparative Example 1 using astaxanthin without special treatment and granulated using conventional high temperature.

[0058] Blank control group: fed with a basal diet without any additional astaxanthin (its formula is similar to the basal diet components of Example 1, but does not contain enzymatically hydrolyzed astaxanthin biomass and microencapsulated astaxanthin, nor does it contain other astaxanthin-derived functional additives for spraying).

[0059] 2. Feeding and Management: Each group of fighting cocks was housed under identical cage conditions and daily management (temperature 20-25°C, relative humidity 55-65%, and 16 hours of light per day). A pre-feeding period of 7 days was performed to acclimate the birds to the experimental diet. The main experimental period lasted 35 days. During the experimental period, the birds had free access to the corresponding group feed and were provided with ample clean drinking water.

[0060] 3. Sample collection and testing: From the 29th to the 35th day of the formal experimental period (7 days in total), all eggs laid by each replicate group were collected every day.

[0061] Five eggs of normal appearance were randomly selected from each replicate group daily. The yolks were completely separated, and the five yolks from the same replicate were combined and gently stirred to create a mixed yolk sample.

[0062] Yolk color scoring: The mixed egg yolk samples were scored for color using the Roche Yolk Color Fan (15 levels).

[0063] Assay for astaxanthin content in egg yolk: Accurately weigh an appropriate amount (approximately 1-2 grams) of mixed egg yolk sample and grind and extract it with an organic solvent (e.g., a mixture of n-hexane and isopropanol). Centrifuge and collect the supernatant. Repeat the extraction several times until the egg yolk residue is colorless. Combine the extracts and concentrate to near dryness under a stream of nitrogen. Dissolve the residue in an appropriate amount of mobile phase, filter, and analyze the astaxanthin content using high-performance liquid chromatography (HPLC) at a specific wavelength (e.g., 474 nm). Quantify the astaxanthin content using a standard. The result is expressed as mg / kg egg yolk.

[0064] 4. Evaluation indicators: Compare the average astaxanthin content (mg / kg egg yolk) and the average Roche colorimetric fan score in each group.

[0065] The experimental data are shown in Table 2: Table 2: Effects of different feeds on astaxanthin content and color of egg-laying poultry egg yolks The test results in Table 2 clearly show that the deposition of astaxanthin in the egg yolks of egg-laying fighting chickens fed with the feed of Example 1 was significantly higher than that of the control group 1 and the blank control group. This is mainly due to the efficient delivery and protection mechanism of astaxanthin in the feed of Example 1. The natural astaxanthin biomass treated with enzymatic hydrolysis improves its release and absorption potential in the digestive tract, while the high-purity astaxanthin coated with liposome-chitosan composite microcapsules effectively resists the damage of digestive juices with its unique dual protection structure, and may significantly improve the bioavailability of astaxanthin by promoting the absorption of intestinal epithelial cells. These specially treated astaxanthin components, combined with the gentle processing technology of low-temperature granulation and vacuum dynamic post-spraying, ensure that more active astaxanthin can be absorbed by the chicken body and effectively transferred to the egg yolk.

[0066] The Roche colorimetric fan score of egg yolk color showed a high positive correlation with the astaxanthin content in the egg yolk. The egg yolk color score of Example 1 group was the highest, indicating that its egg yolk showed a more vivid and ideal orange-red color, which directly reflected the higher astaxanthin concentration in the egg yolk. In contrast, although astaxanthin was also added to the comparative example 1 group, because its astaxanthin source was not specially treated and high-temperature granulation was used, the astaxanthin loss during feed processing and animal digestion was large, and the bioavailability was low. Therefore, the egg yolk astaxanthin deposition amount and color score were much lower than those of the example 1 group. The egg yolk astaxanthin content of the blank control group was extremely low, and the color score was also the lowest, which was in line with expectations.

[0067] Therefore, the present invention combines an enzymatic pretreatment of the astaxanthin source with liposome-chitosan composite microencapsulation, combined with an advanced feed preparation process involving low-temperature granulation and vacuum dynamic post-spraying. This not only ensures the high stability and activity of astaxanthin in the feed, but more importantly, significantly improves its bioavailability in egg-laying fighting chickens and its deposition efficiency in the egg yolk. This results in eggs rich in astaxanthin and exhibiting the yolk color preferred by consumers, fully demonstrating the superiority of the present technical solution in improving the nutritional value and sensory quality of eggs.

[0068] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A high-nutrition egg feed for fighting chickens rich in astaxanthin, characterized in that: Calculated by weight percentage: 1.0-3.0% enzymatic yeast or algae-derived astaxanthin biomass; An astaxanthin product coated with 0.05-0.15% liposome-chitosan composite microcapsules, wherein the astaxanthin encapsulation content in the astaxanthin product is 8-12%; At least one functional additive selected from the following: 0.5-1.5% L-arginine; 0.3-1.0% creatine monohydrate; 0.06-0.15% L-ascorbic acid-2-polyphosphate; 0.02-0.1% of a composite probiotic preparation containing at least one of Bacillus coagulans, Enterococcus faecalis, or Lactobacillus plantarum; and Basic feed matrix of protein source, energy source, vitamins and minerals.

2. The astaxanthin-rich high-nutrition egg feed for fighting chickens according to claim 1, characterized in that The enzymatic yeast or algae-derived astaxanthin biomass is derived from fermented Phaffia rhodozyma or Haematococcus pluvialis.

3. The astaxanthin-rich high-nutrition egg feed for fighting chickens according to claim 1, characterized in that The liposome-chitosan composite microcapsule-encapsulated astaxanthin product is prepared using high-purity astaxanthin with a purity of ≥95%.

4. The astaxanthin-rich high-nutrition egg feed for fighting chickens according to claim 1, characterized in that Also includes at least one functional additive selected from the following: 0.05-0.2% L-carnitine; 0.03-0.06% DL-α-tocopheryl acetate; 0.015-0.03% organic selenium; Biotin added in an amount of 0.2-0.5 mg / kg based on feed; 0.1-0.3% methionine zinc complex; 0.5-1.5% small molecule collagen peptides.

5. The astaxanthin-rich high-nutrition egg feed for fighting chickens and its preparation process according to claim 1 or 4, characterized in that: It also includes 1.0-3.0% fish oil rich in EPA and DHA, 2.0-5.0 mg / kg folic acid added in the feed, and 0.02-0.05 mg / kg vitamin B12 added in the feed.

6. A process for preparing the high-nutrition egg feed for fighting chickens rich in astaxanthin according to any one of claims 1 to 5, characterized in that: The following steps are involved: (i) preparing enzymatic yeast or algae-derived astaxanthin biomass; (ii) preparing an astaxanthin product coated with liposome-chitosan composite microcapsules; (iii) uniformly mixing the basic feed components with the enzymatic yeast or algal astaxanthin biomass obtained in step (i) and any other heat-resistant additives, and performing low-temperature granulation to form basic granules, wherein the granule temperature does not exceed 65° C. during the granulation process; (iv) vacuum dynamically coating the base particles obtained in step (iii) by spraying a liquid mixture comprising the astaxanthin product encapsulated by the liposome-chitosan composite microcapsules obtained in step (ii) and other thermosensitive active ingredients under vacuum conditions of -0.05 to -0.08 MPa relative pressure.

7. The preparation process according to claim 6, characterized in that: Step (i) comprises: mixing yeast or algae-derived astaxanthin biomass with a buffer solution having a pH of 4.5 to 5.5, adding a composite enzyme preparation accounting for 0.5 to 2.0% of the dry weight of the biomass, hydrolyzing at 45 to 55° C. for 2 to 6 hours, and then inactivating the enzyme and drying.

8. The preparation process according to claim 6, characterized in that: Step (ii) comprises: dissolving high-purity astaxanthin, phosphatidylcholine and cholesterol to form a lipid membrane, hydrating to form a coarse liposome suspension, and then ultrasonically treating the suspension at a power of 100 to 300 W for 5 to 20 minutes to form nano-astaxanthin liposomes; then dropwise adding the nano-astaxanthin liposomes into a 0.1 to 0.5% w / v chitosan solution with a pH of 4.0 to 5.5 for coating, and collecting and drying the microcapsules.

9. The preparation process according to claim 6, characterized in that: The low-temperature granulation in step (iii) is cold-pressed granulation, the temperature of the material before entering the granulator die does not exceed 40°C, and the granule discharge temperature does not exceed 60-65°C.

10. The preparation process according to claim 6, characterized in that: The vacuum dynamic coating in step (iv) is carried out at a temperature of 20-35° C., and the amount of the liquid mixture sprayed is 2-5% of the total weight of the base particles.

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