Efficient fattening type compound feed for meat gamecocks and preparation method of efficient fattening type compound feed

Abstract

The invention relates to the field of animal feed science and processing, and discloses an efficient fattening type compound feed for meat gamegalors and a preparation method thereof.The preparation method includes the steps that firstly, specific compound plant raw materials and screened compound dominant flora are adopted for multi-step solid-state symbiotic fermentation, and a fermentation base material rich in probiotics, small molecule peptides and low anti-nutritional factors is prepared; secondly, core functional components including high-activity probiotics, plant extracts, functional lipids, antioxidants and the like are subjected to targeted sustained-release microcapsule coating through a multi-layer composite material; and finally, mixing the fermentation base material with targeted slow-release microcapsules and other conventional feed components, and preparing the compound feed by adopting a low-temperature granulation process. The nutritional value and the utilization rate of the feed can be remarkably improved, and functional components are effectively protected and delivered in a targeted manner, so that the growth and development of the meat gamecocks are promoted, the immune function and the oxidation resistance of the meat gamecocks are enhanced, the meat quality is improved, and the overall health level is improved.

Description

Technical Field

[0001] The invention relates to the field of animal feed science and processing technology, in particular to a high-efficiency fattening compound feed for broiler fighting chickens and a preparation method thereof. Background Art

[0002] As a specialty poultry, broiler fighting chickens have attracted market attention due to their unique flavor and nutritional value. In order to improve the production performance and meat quality of broiler fighting chickens, the scientific formulation and processing of feed are crucial. Currently, in the research and development and application of broiler fighting chicken feed, researchers are committed to improving feeding results by optimizing feed formulas and improving processing technology. For example, in order to reduce feed costs and make full use of resources, plant protein raw materials such as rapeseed meal and soybean meal are widely used. However, the inherent anti-nutritional factors in these raw materials, such as glucosinolates, trypsin inhibitors, phytic acid, etc., often limit the full realization of their nutritional value, affect the digestion and absorption of nutrients by fighting chickens, and may even have an adverse effect on intestinal health.

[0003] To enhance fighting cocks' immunity, promote growth, and improve meat quality, various functional additives are often added to their feed, such as probiotics, plant extracts, functional lipids, and specific vitamins and trace elements. However, the stability and bioactivity of these functional components, particularly the activity of probiotics, active ingredients in plant extracts, and easily oxidized unsaturated fatty acids and vitamins, are often severely affected by environmental factors (such as oxygen, humidity, temperature, and light) and physical forces (such as shear force and pressure) during feed storage, transportation, and pelleting. This results in the actual effective dose entering the animal's body being far lower than expected, making it difficult for them to fully exert their expected physiological effects.

[0004] In addition, existing feed processing technology, especially the conventional high-temperature pelleting process, although it can improve the palatability and physical form of the feed, may also lead to a further reduction in the activity of heat-sensitive nutrients and functional additives in the feed. Even if certain functional components can withstand the processing process, their release in the animal's digestive tract is often lacking in targeting. They may be digested, degraded or absorbed before reaching the target site of action (such as the hindgut), which greatly reduces their regulatory effect on specific physiological functions. Therefore, how to effectively protect the key functional components in the feed, ensure their stability and bioactivity, and achieve their precise targeted release in the digestive tract, while optimizing the nutritional value of the basic feed raw materials, is a technical problem that needs to be solved urgently in the current field of broiler fighting chicken feed, which is directly related to the healthy breeding of fighting chickens, the improvement of production efficiency and the quality of the final product. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the present invention aims to solve the technical problems of low utilization rate of plant raw materials, easy inactivation and lack of targeting of functional additives in the existing compound feed for meat - use fighting cocks, resulting in poor overall feeding effect.

[0006] To achieve the above - mentioned purpose, the present invention is realized through the following technical solutions: A high - efficiency fattening compound feed for meat - use fighting cocks, by mass fraction, includes: 20 - 40 parts of fermentation base material; the fermentation base material is prepared by multi - stage solid - state symbiotic fermentation of a composite substrate by a core fermentation flora. The composite substrate contains, by dry matter weight percentage, 30 - 40% of rapeseed meal, 20 - 30% of pretreated fermented soybean meal, 15 - 25% of corn germ meal, and 10 - 20% of composite fruit and vegetable processing by - products; the core fermentation flora includes detoxifying Bacillus, Aspergillus niger, Lactobacillus plantarum and Saccharomyces cerevisiae; Other conventional feed components, supplemented to 100 parts, and the other conventional feed components are selected from one or more of energy feed, protein feed, mineral feed, vitamin premix and trace element premix.

[0007] These conventional components provide basic macro - and micro - nutrients for fighting cocks, ensuring the comprehensiveness and balance of feed nutrition.

[0008] Preferably, the composite fruit and vegetable processing by - products in the fermentation base material are a mixture of apple pomace, tomato pomace and citrus peel in a weight ratio of 1:1:0.5.

[0009] Preferably, the core fermentation flora of the fermentation base material specifically includes detoxifying Bacillus strain XJ - 5, Aspergillus niger strain FZ - 2, Lactobacillus plantarum strain DG - 8 and Saccharomyces cerevisiae strain YM - 3.

[0010] Preferably, in the plant extract complex in the core component A of the targeted slow - release microcapsule, the weight ratio of astragalus polysaccharide in astragalus extract, icariin in epimedium extract and atractylenolide II in atractylodes macrocephala extract is 5:1:0.5.

[0011] Preferably, in the core component B of the targeted slow - release microcapsule, the weight ratio of α - linolenic acid to conjugated linoleic acid is 2.5:1, and the ratio of natural vitamin E to selenium element in organic selenium is 100 IU:1 mg Se.

[0012] Preferably, the at least double - layer coating structure of the targeted slow - release microcapsule further includes an enzyme - sensitive material to achieve the programmed targeted release of core component A and core component B in the digestive tract of fighting cocks.

[0013] Preferably, a method for preparing a high - efficiency fattening compound feed for meat - use fighting cocks is characterized by including the following steps: S1. Preparation of fermentation base material: A composite substrate containing rapeseed meal, pretreated fermented soybean meal, corn germ meal, and composite fruit and vegetable processing by-products is subjected to multi-step solid-state symbiotic fermentation with a core fermentation flora, which includes Bacillus cereus for detoxification, Aspergillus niger, Lactobacillus plantarum, and Saccharomyces cerevisiae; the multi-step solid-state symbiotic fermentation includes the step of dynamically inoculating subsequent dominant strains based on intermediate metabolites or enzyme activity indexes during the fermentation process; S2. Preparation of targeted slow-release microcapsules: Core component A containing Clostridium butyricum and a plant extract complex, and core component B containing α-linolenic acid, conjugated linoleic acid, natural vitamin E, and organic selenium are subjected to at least double-layer coating treatment, and the coating treatment includes using a pH-sensitive enteric material; S3. The fermentation base material obtained in step S1, the targeted slow-release microcapsules obtained in step S2, and other conventional feed components are mixed evenly to obtain the composite feed.

[0014] Preferably, in step S1 of preparing the fermentation base material, the multi-step solid-state symbiotic fermentation specifically includes: S1-1. First-stage fermentation: Under aerobic conditions at a temperature of 28-30 °C and a material humidity of 56-60%, Aspergillus niger and Bacillus cereus for detoxification are inoculated for fermentation for 48-72 hours; S1-2. Second-stage fermentation: When the key enzyme activity or the concentration of characteristic hydrolysis products in the first-stage fermentation reaches a preset threshold, Lactobacillus plantarum and Saccharomyces cerevisiae are inoculated, and fermentation is carried out for 72-96 hours under microaerobic or anaerobic conditions at a temperature of 26-28 °C and a material humidity of 51-55%.

[0015] Preferably, in step S2 of preparing the targeted slow-release microcapsules, the at least double-layer coating treatment further includes coating with an enzyme-sensitive material, and the enzyme-sensitive material is selected from at least one of starch acetate with a specific degree of substitution or modified pectin; through the combined use of the pH-sensitive enteric material and the enzyme-sensitive material, programmed targeted release of core component A and core component B in different sections of the fighting cock digestive tract is achieved.

[0016] Preferably, after step S3, there are also tempering and low-temperature granulation steps; the tempering step is to temper the evenly mixed material at 60-68 °C under a steam pressure of 0.2-0.3 MPa for 60-90 seconds; the low-temperature granulation step is to granulate the tempered material under the condition that the temperature of the granule exiting the die is ≤70-75 °C.

[0017] The present invention provides a high-efficiency fattening composite feed for meat-type fighting cocks and a preparation method thereof. It has the following beneficial effects: 1. By using a pre-treated composite plant-based feedstock as a fermentation base and combining it with a selected composite dominant bacterial community for multi-step solid-state symbiotic fermentation, the present invention significantly degrades anti-nutritional factors in the feedstock, improves the digestibility and utilization of nutrients, and produces a rich supply of probiotics, small molecule peptides, and other beneficial metabolites. This optimized fermentation base improves the overall quality of the feed, thereby effectively promoting the growth and development of broiler fighting chickens, increasing feed conversion, and laying a solid nutritional foundation for the healthy fattening of fighting chickens.

[0018] 2. This invention innovatively encapsulates specific immune-enhancing and growth-promoting ingredients (such as highly active Clostridium butyricum and specific plant extracts) and meat-quality-improving and antioxidant ingredients (such as functional lipids and highly effective antioxidants) using multi-layer targeted sustained-release microencapsulation technology. This sophisticated encapsulation structure effectively protects these core active ingredients from damage during feed processing, storage, and the upper digestive tract, while enabling programmed release in specific areas of the intestine. This significantly improves the bioavailability and efficacy of these functional ingredients, specifically enhancing the immune function and meat quality of broiler fighting chickens.

[0019] 3. The overall formula design of the compound feed of this invention combines the dual advantages of an optimized fermentation base and targeted sustained-release microencapsulation technology. A gentle, low-temperature granulation process ensures the stability of various heat-sensitive active ingredients in the feed. This ensures that the compound feed not only fully meets the nutritional needs of broiler fighting chickens during their fattening period, but also significantly enhances their overall health and disease resistance by improving intestinal microecological balance, enhancing antioxidant capacity, and strengthening immune defenses. This results in the production of a safe, high-quality fighting chicken product with promising application prospects and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The present invention is a flow chart of the preparation method of high-efficiency fattening compound feed for broiler fighting chickens. DETAILED DESCRIPTION

[0021] The following will clearly and completely describe the technical solution of the present invention in conjunction with the accompanying drawings. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0022] Please see the attached Figure 1 Example: Example 1 1. Preparation of fermentation base Raw material preparation and pretreatment: Composition of composite substrate (by dry matter weight): rapeseed meal (low glucosinolate variety, dehulled) 35%, pre-treated fermented soybean meal (pre-fermented by lactic acid bacteria) 25%, corn germ meal 20%, composite fruit and vegetable processing by-products (apple pomace, tomato pomace, citrus peel mixed in a weight ratio of 1:1:0.5 and dried) 20%.

[0023] Ultra-finely grind rapeseed meal to D90 < 150 μm; separately grind corn germ meal and composite fruit and vegetable processing by-products through a 60-mesh sieve.

[0024] Dry mix the above-ground raw materials with pre-treated fermented soybean meal in a V-type mixer for 12 minutes.

[0025] Add the mixed material to a solid-state fermentation tank, introduce clean steam to adjust the moisture content of the material to 58% (w / w), and perform moist heat sterilization at 118 °C for 20 minutes. After sterilization, cool the material to 30 °C.

[0026] Strain activation and dynamic inoculation: Core fermentation flora: Bacillus subtilis strain XJ-5, Aspergillus niger strain FZ-2, Lactobacillus plantarum strain DG-8, Saccharomyces cerevisiae strain YM-3. Each strain is separately activated and cultured to prepare a bacterial suspension with a viable bacteria concentration ≥ 1x10 9 CFU / mL.

[0027] Initial inoculation: Aseptically inoculate the black Aspergillus bacterial suspension (inoculation amount is 0.7% v / w of the dry weight of the fermented material) and the Bacillus subtilis bacterial suspension (inoculation amount is 0.7% v / w of the dry weight of the fermented material) into the sterilized and cooled substrate.

[0028] Process monitoring and secondary inoculation trigger: During the first-stage fermentation process, monitor the cellulase activity and reducing sugar content. When the cellulase activity reaches 75% of the peak value and the reducing sugar content reaches the preset range, start the second-stage strain inoculation.

[0029] Multi-step solid-state symbiotic fermentation: First stage: After inoculation, set the fermentation temperature at 29 °C, the initial material humidity at 58%, aerobic conditions (sterile air ventilation volume 0.35 vvm), stir once every 3 hours (rotation speed 8 rpm), and ferment for 60 hours.

[0030] Second stage: After reaching the triggering condition for secondary inoculation, add a sterilized glucose solution equivalent to 6% (w / w) of the dry weight of the fermented material. Inoculate aseptically with an activated Lactobacillus plantarum bacterial suspension (inoculation amount: 1.2% v / w of the dry weight of the fermented material) and a Saccharomyces cerevisiae bacterial suspension (inoculation amount: 1.2% v / w of the dry weight of the fermented material). Adjust the fermentation temperature to 27°C, maintain the material humidity at 53%, switch to micro-aerobic fermentation (sterile air ventilation rate: 0.08 vvm), stir once every 7 hours, and ferment for 84 hours.

[0031] Post-treatment: After fermentation, the fermented material is vacuum freeze-dried at a temperature ≤50°C to make the moisture content of the final product ≤9% (w / w).

[0032] After cooling, the dried fermented material is crushed to pass through a 50-mesh sieve. The finished product is vacuum-packed and stored in the dark at 8°C.

[0033] II. Preparation of targeted sustained-release microcapsules Pretreatment and mixing of core components A and B: Core component A: Mix the bacterial powder of highly active Clostridium butyricum (derived from the healthy intestine of fighting cocks, viable count ≥1x10 10 CFU / g) with a plant extract complex (weight ratio of astragalus polysaccharide: icariin: atractylenolide II is 5:1:0.5), together with trehalose (7% w / w of the dry weight of the core material) and hydroxypropyl-β-cyclodextrin (15% w / w of the dry weight of the plant extract) evenly.

[0034] Core component B: Thoroughly mix a mixture of α-linolenic acid and conjugated linoleic acid (weight ratio 2.5:1) with natural vitamin E and high-purity yeast selenium (the ratio of selenium element in natural vitamin E to organic selenium is 100 IU:1 mg Se), together with lecithin and sucrose fatty acid ester (weight ratio 4:1, 5% w / w of the total weight of the oil phase) and ascorbyl palmitate (0.03% w / w of the total weight of the oil phase).

[0035] Preparation of the inner-layer microcapsules: Core component A: Using coaxial air-assisted electrospray technology, with chitosan at 2.0% w / v as the shell material and sodium alginate at 1.5% w / v as the cross-linking agent. Spray voltage: 25 kV, liquid flow rate: 1.0 mL / h, receiving distance: 12 cm.

[0036] Core component B: Dispersing the oil phase in an aqueous phase containing a stabilizer to form an O / W emulsion by high-pressure homogenization (100 MPa, 4 cycles), and then preparing microcapsules by spray drying technology (inlet temperature: 145°C, outlet temperature: 70°C).

[0037] Outer-layer composite coating: Using the inner microcapsules as the core, coating was carried out using a Wurster bottom spray fluidized bed coater.

[0038] Coating material solution: For the first pH-sensitive layer: Eudragit® L100-55 and S100 were mixed at 1:1 (w / w) and dissolved in an 85:15 (v / v) ethanol / water system with a polymer concentration of 7% (w / v); for the second enzyme-sensitive layer: starch acetate with a degree of substitution of 1.0 was dissolved in water with a polymer concentration of 5% (w / v).

[0039] Coating process parameters: Inlet air temperature 48°C, material temperature 35°C, atomization pressure 0.15 MPa, liquid spraying rate 10 g / min·kg of core, and the total coating weight gain was controlled at 18% (w / w) of the core weight.

[0040] Obtaining the finished microcapsules: After coating, fluidized drying was carried out at 40°C for 1.5 hours. Precision screening was carried out through a 150 μm sieve, and microcapsules with a particle size of about 100 - 300 μm were collected. Vacuum packaging was carried out and stored in the dark and dry at 8°C.

[0041] III. Preparation of the final compound feed Weighing and mixing: By mass fraction, 30 parts of the fermented base material were weighed, 0.3 parts of the targeted slow-release microcapsules, and the rest were made up to 100 parts by conventional feed components such as corn, extruded soybean, calcium hydrogen phosphate, stone powder, salt, vitamin premix, and trace element premix.

[0042] Premixing: The vitamin premix, trace element premix, and targeted slow-release microcapsules were pre-mixed in gradients with corn flour 8 times their total weight.

[0043] Main mixing: A horizontal screw ribbon mixer was used, and each component was added in sequence according to the principle of "from more to less". The total mixing time was 4 minutes to ensure that the coefficient of variation of the mixing uniformity CV ≤ 5%.

[0044] Conditioning and granulation: The mixed powder entered the conditioner, with a conditioning temperature of 65°C, steam pressure of 0.25 MPa, and conditioning time of 75 seconds. Granulation was carried out using a ring die granulator, with a particle diameter of 3.5 mm, a ring die compression ratio of 10:1, and the temperature of the particles leaving the die was controlled at 72°C.

[0045] Cooling and packaging: The pellet feed entered a countercurrent cooler and was cooled to the ambient temperature + 4°C, with a moisture content of 12% (w / w). The finished product was sealed and packaged using multi-layer composite moisture-proof and light-proof packaging bags.

[0046] Example 2 I. Preparation of the fermented base material Raw material preparation and pretreatment: Composition of composite substrate (by dry matter weight): rapeseed meal 30%, pretreated fermented soybean meal 20%, corn germ meal 15%, composite fruit and vegetable processing by-products 10%.

[0047] The raw materials are ground to basically pass through a 60-mesh sieve.

[0048] The mixed materials are dry-mixed in a mixer for 10 minutes.

[0049] The moisture of the mixed materials is adjusted to 56% (w / w), and heat-moisture sterilization is carried out at 115 °C for 18 minutes. Cool to 28 °C.

[0050] Activation of strains and dynamic inoculation: The core fermentation flora is the same as in Example 1.

[0051] Initial inoculation: Suspensions of Aspergillus niger and Bacillus detoxificans are each inoculated at 0.5% v / w of the dry weight of the fermented material.

[0052] The triggering conditions for secondary inoculation are the same as in Example 1.

[0053] Multi-step solid-state symbiotic fermentation: The first stage: The fermentation temperature is 28 °C, the initial moisture of the material is 56%, aerobic conditions (aeration rate 0.2 vvm), stirring once every 4 hours (rotation speed 5 rpm), and fermentation for 48 hours.

[0054] The second stage: After reaching the triggering conditions for secondary inoculation, no additional sugar source is added. Suspensions of Lactobacillus plantarum and Saccharomyces cerevisiae after activation are each inoculated at 0.8% v / w of the dry weight of the fermented material. Adjust the fermentation temperature to 26 °C, maintain the moisture of the material at 51%, and switch to anaerobic fermentation (introduce N2 for protection, no stirring), and ferment for 72 hours.

[0055] Post-treatment: The fermented material is dried by low-temperature air flow puffing and dried at ≤55 °C to make the moisture content of the final product ≤10% (w / w).

[0056] The dried fermented material is ground to pass through a 40-mesh sieve. Packed with nitrogen filling and stored in the dark at 10 °C.

[0057] II. Preparation of targeted sustained-release microcapsules Pretreatment and mixing of core components A and B: Core component A: The complex of Clostridium butyricum powder and plant extract (the component ratio is the same as in Example 1), is blended with trehalose (5% w / w of the dry weight of the core substance) and hydroxypropyl-β-cyclodextrin (10% w / w of the dry weight of the plant extract).

[0058] Core Component B: α-linolenic acid and conjugated linoleic acid (weight ratio same as in Example 1), natural vitamin E and yeast selenium (formulation ratio same as in Example 1), are mixed with lecithin and sucrose fatty acid ester (weight ratio 3:1, accounting for 3% w / w of the total oil phase weight) and ascorbyl palmitate (accounting for 0.01% w / w of the total oil phase weight).

[0059] Preparation of the inner microcapsules: Core Component A: Using the ion gelation method, chitosan at 1.5% w / v is used as the shell material, and sodium alginate at 1.0% w / v is used as the cross-linking agent.

[0060] Core Component B: Microcapsules are prepared by the membrane emulsification-solvent evaporation method (membrane pore size 0.5 μm).

[0061] Outer composite coating: A fluidized bed coater is used.

[0062] Coating material solution: The first layer is a pH-sensitive layer: Eudragit® L100-55, dissolved in a 90:10 (v / v) ethanol / water system, with a polymer concentration of 5% (w / v); the second layer is an enzyme-sensitive layer: low-methoxyl pectin, dissolved in water, with a polymer concentration of 3% (w / v).

[0063] Coating process parameters: Inlet air temperature 40°C, material temperature 30°C, atomization pressure 0.1 MPa, liquid spraying rate 5 g / min·kg of core material, and the total coating weight gain is controlled at 10% (w / w) of the core material weight.

[0064] Obtaining the finished microcapsules: After coating, fluidized drying is carried out at 35°C for 1 hour. Screening is carried out through a 200-mesh sieve, and microcapsules with a particle size of about 75 - 100 μm are collected. Vacuum packaging is carried out, and storage is carried out in the dark and dried at 10°C.

[0065] III. Preparation of the final compound feed Weighing and mixing: By mass fraction, 20 parts of the fermentation base material and 0.1 part of the targeted slow-release microcapsules are weighed, and the rest is made up to 100 parts with conventional feed components.

[0066] Premixing: The trace components are premixed with a carrier that is 5 times their total weight.

[0067] Main mixing: The total mixing time is 3 minutes, and CV ≤ 5%.

[0068] Conditioning and pelletizing: Conditioning temperature 60°C, steam pressure 0.2 MPa, conditioning time 60 seconds. The compression ratio of the pellet mill ring die is 8:1, and the temperature of the pellets leaving the die is 70°C.

[0069] Cooling and packaging: Cooling to the ambient temperature + 5°C, with a moisture content of 12.5% (w / w).

[0070] Example 3 I. Preparation of Fermentation Substrate Raw Material Preparation and Pretreatment: Composition of Composite Substrate (by dry matter weight): rapeseed meal 40%, pretreated fermented soybean meal 30%, corn germ meal 25%, composite fruit and vegetable processing by - products 20%.

[0071] The requirement for raw material crushing is the same as that in Example 1.

[0072] The mixed materials are dry - mixed in a mixer for 15 minutes.

[0073] The moisture of the mixed materials is adjusted to 60% (w / w), and they are sterilized by moist heat at 120 °C for 22 minutes. Then they are cooled to 32 °C.

[0074] Activation of Bacteria Strains and Dynamic Inoculation: The core fermentation bacterial community is the same as that in Example 1.

[0075] Initial inoculation: Aspergillus niger and Bacillus detoxification bacterial suspensions are each inoculated at 1.0% v / w of the dry weight of the fermentation material.

[0076] Trigger condition for secondary inoculation: Start when the cellulase activity reaches 80% of the peak value and the reducing sugar content reaches the preset upper limit.

[0077] Multi - stage Solid - state Symbiotic Fermentation: First stage: Fermentation temperature is 30 °C, initial material humidity is 60%, aerobic condition (aeration rate 0.5 vvm), continuous low - speed stirring (rotation speed 10 rpm), fermentation for 72 hours.

[0078] Second stage: After reaching the secondary inoculation trigger condition, add sterilized molasses at 8% (w / w) of the dry weight of the fermentation material. Activated Lactobacillus plantarum and Saccharomyces cerevisiae bacterial suspensions are each inoculated at 1.5% v / w of the dry weight of the fermentation material. Adjust the fermentation temperature to 28 °C, maintain the material humidity at 55%, turn to micro - aerobic fermentation (aeration rate 0.1 vvm), stir once every 6 hours, and ferment for 96 hours.

[0079] Post - treatment: The fermentation material is freeze - dried under vacuum and dried at ≤55 °C to make the moisture content of the final product ≤8% (w / w).

[0080] The dried fermentation material is crushed to pass through a 60 - mesh sieve. It is vacuum - packaged and stored in the dark at 4 °C.

[0081] II. Preparation of Targeted Sustained - Release Microcapsules Pretreatment and Mixing of Core Components A and B: Core Component A: A complex of Clostridium butyricum powder and plant extracts (with the same component ratio as in Example 1), blended with glycerol and lactose (total addition amount accounting for 10% w / w of the dry weight of the core material) and hydroxypropyl-β-cyclodextrin (accounting for 20% w / w of the dry weight of the plant extracts).

[0082] Core Component B: α-linolenic acid and conjugated linoleic acid (with the same weight ratio as in Example 1), natural vitamin E and yeast selenium (with the same ratio as in Example 1), mixed with lecithin and sucrose fatty acid ester (weight ratio 5:1, accounting for 7% w / w of the total weight of the oil phase) and ascorbyl palmitate (accounting for 0.05% w / w of the total weight of the oil phase).

[0083] Preparation of the inner microcapsules: Core Component A: Using coaxial air-assisted electrostatic spraying technology, with chitosan at 2.5% w / v as the shell material and sodium alginate at 2.0% w / v as the cross-linking agent. The spraying voltage is 30 kV, the liquid flow rate is 1.5 mL / h, and the receiving distance is 15 cm.

[0084] Core Component B: After high-pressure homogenization (120 MPa, 5 cycles), microcapsules are prepared by spray cooling technology (inlet temperature 160 °C, outlet temperature 80 °C).

[0085] Outer composite coating: Using a Wurster bottom-spray fluidized bed coater.

[0086] Coating material solution: The first layer is a pH-sensitive layer: Eudragit® L100-55 and S100 are mixed at 1:2 (w / w), dissolved in an 80:20 (v / v) ethanol / water system, and the polymer concentration is 10% (w / v); the second layer is an enzyme-sensitive layer: modified pectin, dissolved in a buffer solution, and the polymer concentration is 8% (w / v); an optional third layer is a pH highly sensitive layer: Eudragit® FS30D aqueous dispersion.

[0087] Coating process parameters: Inlet air temperature 55 °C, material temperature 40 °C, atomization pressure 0.2 MPa, liquid spraying rate 15 g / min·kg of the core, and the total coating weight gain is controlled at 25% (w / w) of the core weight.

[0088] Obtaining the finished microcapsules: After coating, fluidized drying is carried out at 45 °C for 2 hours. Screening is carried out through an 80-mesh sieve, and microcapsules with a particle size of about 180 - 500 μm are collected. Vacuum packaging and storage in the dark at 4 °C.

[0089] III. Preparation of the final compound feed Weighing and mixing: By mass fraction, 40 parts of the fermentation base material and 1.0 part of the targeted sustained-release microcapsules are weighed, and the rest is made up to 100 parts by conventional feed components.

[0090] Premixing: The trace components are premixed with a carrier whose total weight is 10 times that of the trace components.

[0091] Main mixing: A high-efficiency double-shaft paddle mixer is used, and the total mixing time is 5 minutes, with CV ≤ 5%.

[0092] Conditioning and pelletizing: The conditioning temperature is 68 °C, the steam pressure is 0.3 MPa, and the conditioning time is 90 seconds. The pellet diameter is 4.5 mm, the ring die compression ratio is 12:1, and the pellet outlet temperature is 75 °C.

[0093] Cooling and packaging: Cooled to the ambient temperature + 3 °C, with a moisture content of 11% (w / w). The finished product is sealed and packaged in a multi-layer composite moisture-proof and light-proof packaging bag.

[0094] Comparative example: Comparative example 1 Compared with Example 1, the differences are as follows: The preparation of the fermentation base material is different: Simplification of the composite substrate and no pretreatment: The composite substrate used in its fermentation base material consists of: ordinary rapeseed meal 35% (without low glucosinolate screening and peeling treatment), ordinary soybean meal 25% (without pretreatment fermentation), corn germ meal 20%, and ordinary wheat bran 20% (replacing the composite fruit and vegetable processing by-products). All raw materials are only subjected to conventional crushing.

[0095] Simplification of the fermentation process: Instead of using the multi-step solid-state symbiotic fermentation and dynamic inoculation process, the simplified composite substrate described above and Aspergillus niger (FZ-2 strain), a single strain in the core fermentation flora, are subjected to single-stage solid-state fermentation for 72 hours at 28 - 30 °C and a material humidity of 55 - 60%. During this period, no other strains are inoculated, and there is no specific monitoring of enzyme activity or metabolites for regulation.

[0096] The remaining components, the preparation of the targeted slow-release microcapsules (if still used), and the preparation method of the final compound feed (except for the different source of the fermentation base material) are the same as those in Example 1.

[0097] Comparative example 2 Compared with Example 1, the differences are as follows: The treatment method of the targeted slow-release microcapsules is different: No microcapsule coating: The targeted slow-release microcapsules are not prepared. The core component A (complex of Clostridium butyricum and plant extracts) and core component B (combination of α-linolenic acid, conjugated linoleic acid, natural vitamin E, and organic selenium) contained in the original targeted slow-release microcapsules in Example 1 are directly mixed with other feed components in their original, uncoated form, at the same addition amount as in Example 1 (0.3 parts by mass, referring to the total dry weight of core components A and B).

[0098] The preparation of the fermentation base material, the selection of other conventional feed components, and the preparation method of the final compound feed (except for the different treatment methods of the microcapsule part) are the same as those in Example 1.

[0099] Comparative Example 3 Compared with Example 1, the differences are as follows: The coating structure of the targeted sustained-release microcapsule is simplified: Although microcapsules are still prepared, their coating structure is significantly simplified. After the preparation of the inner-layer microcapsules of Core Component A and Core Component B, only a single ordinary coating material (such as gelatin or gum arabic, without specific pH-sensitive or enzyme-sensitive properties) is used for single-layer coating, and the coating weight gain is only 5% of the weight of the capsule core. A multi-layer targeted release structure composed of pH-sensitive enteric materials and enzyme-sensitive materials is not used.

[0100] The granulation process of the final compound feed is different: After final uniform mixing, a conventional high-temperature granulation process is adopted. The temperature in the conditioning step is increased to 85 °C, the steam pressure is 0.35 MPa, and the conditioning time is 30 seconds; the die temperature of the granulator and the temperature of the particles leaving the die are both controlled at 85 - 90 °C.

[0101] The preparation of the fermentation base material, the selection of the microcapsule core components, the selection of other conventional feed components, and the mixing method (except for the different microcapsule coating structures and granulation processes) are the same as those in Example 1.

[0102] Test Example: Test Example 1: 1. Experimental Purpose The purpose of this experiment is to verify the innovation and superiority of the preparation process and composition of the fermentation base material in the present invention by comparing the effects of feeding compound feeds containing the innovative fermentation base material (derived from Example 1) and the simplified fermentation base material (derived from Comparative Example 1) on the early growth performance of meat-type fighting cocks, some intestinal health indicators, and the physical and chemical properties of the fermentation base material itself.

[0103] 2. Experimental Design and Animal Grouping Experimental animals: 80 healthy 1-day-old male meat-type fighting cocks (of the same breed and similar body weight) were selected and randomly divided into 2 treatment groups, with 4 replicates in each group and 10 chickens in each replicate.

[0104] Group 1 (CON, control group): Fed a compound feed prepared based on Comparative Example 1 (containing a simplified fermentation base material); Group 2 (EXP, experimental group): Fed a compound feed prepared based on Example 1 (containing an innovative fermentation base material).

[0105] Feeding management: The experimental chickens were raised in a three-dimensional cage under the same environmental conditions (temperature, humidity, and lighting regime were controlled according to the standard procedures), with free access to food and water. The experimental period was 21 days. The immunization program was carried out routinely.

[0106] Feed preparation: CON group feed: The fermented substrate was prepared according to the method of Comparative Example 1, and then mixed evenly with other conventional feed components (corn, extruded soybean, calcium hydrogen phosphate, stone powder, salt, vitamin premix, and trace element premix, whose types and addition ratios were the same as those of the components except the fermented substrate and microcapsules in Example 1), and then conditioned and pelletized at low temperature (the pelletizing parameters were the same as those in Example 1).

[0107] EXP group feed: The compound feed was prepared according to the method of Example 1.

[0108] In both feeds, the addition ratio of the fermented substrate was set at 30% (by mass), and no targeted slow-release microcapsules were added to exclude the influence of microcapsules and focus on comparing the effects of the fermented substrate. The types and ratios of other conventional feed components remained the same.

[0109] 3. Experimental procedures and index determination 3.1 Physicochemical property analysis of the fermented substrate Sample collection: 500 g of samples were randomly taken from the innovative fermented substrate prepared according to the method of Example 1 and the simplified fermented substrate prepared according to the method of Comparative Example 1 respectively, and after mixing evenly, they were used for analysis.

[0110] Determination of antinutritional factors: Glucosinolate content: Referring to the national standard method (such as GB / TXXXX, the specific method depends on the actual detection standard), the total glucosinolate content from rapeseed meal was determined by high performance liquid chromatography (HPLC). Trypsin inhibitor activity: Referring to the national standard method (such as GB / T5009.229), the trypsin inhibitor activity from soybean meal was determined by spectrophotometry.

[0111] Determination of beneficial metabolites: Small molecule peptide content: The total nitrogen was determined by the Kjeldahl method, and the soluble protein was determined by the Folin-Ciocalteu method. The difference between the two was subtracted or evaluated by a specific peptide chain length distribution analysis method (such as gel exclusion chromatography).

[0112] Lactic acid content: It was determined by high performance liquid chromatography (HPLC) or enzyme immunoassay.

[0113] Viable count of Lactobacillus plantarum: An appropriate amount of the fermented substrate sample was taken, serially diluted with sterile saline, spread on MRS agar medium, and the colonies were counted after anaerobic incubation at 37 °C for 48 hours.

[0114] 3.2 Effects of Early Feeding Trials on Growth Performance Data recording: On the 1st and 21st days of the trial, the fasting weights of all fighting cocks were weighed by replicate, and the feed addition and remaining amounts of each replicate were recorded.

[0115] Index calculation: Average daily gain (ADG, g / d / bird) = (total weight at the end of the trial - total weight at the beginning of the trial) / (number of trial days × number of chickens). Average daily feed intake (ADFI, g / d / bird) = total feed consumption / (number of trial days × number of chickens) Feed conversion ratio (FCR) = total feed consumption / total weight gain 3.3 Effects of Early Feeding Trials on Intestinal Health Indicators Sample collection: On the 21st day of the trial, 2 fighting cocks of average weight were randomly selected from each replicate. After sacrificing by carotid artery exsanguination, cecal contents were aseptically collected. One part was immediately stored at -80 °C for microbiota analysis, and the other part was used for VFA determination.

[0116] Volatile fatty acid (VFA) determination: Take 0.5 g of cecal contents, add 1 mL of 25% metaphosphoric acid solution, vortex and mix well, let stand at 4 °C for 30 minutes, and centrifuge at 12000 rpm for 10 minutes. Take the supernatant and filter it through a 0.22 μm filter membrane.

[0117] The concentrations of main VFAs such as acetic acid, propionic acid, and butyric acid were determined by gas chromatography (GC).

[0118] Determination of relative abundances of Lactobacillus plantarum and Clostridium butyricum (qPCR): Extract total DNA from cecal contents.

[0119] Use specific primers targeting the 16S rRNA gene of Lactobacillus plantarum and specific functional genes of Clostridium butyricum (such as butyryl-CoA transferase gene), as well as universal bacterial primers, for real-time fluorescence quantitative PCR.

[0120] Calculate the relative abundances of target microbiota relative to total bacteria (using the 2 -ΔΔCt method or absolute quantification method).

[0121] 4. Statistical Analysis All data were analyzed using SPSS statistical software. Inter-group comparisons were performed using independent sample t-tests or one-way analysis of variance (ANOVA), and the results were expressed as mean ± standard deviation (Mean ± SD). A P value < 0.05 was considered statistically significant.

[0122] 5. Experimental Data Table 1: Effects of Different Fermentation Substrates on the Growth Performance of Fighting Cocks in the Early Stage (1 - 21 Days Old), Main Physicochemical Indexes of Fermentation Substrates, and Contents of Cecal VFA The experimental results clearly show that the quality of the fermentation substrate prepared by using the specific composite substrate and core fermentation flora described in the present invention and through the multi-step solid-state symbiotic fermentation process is significantly optimized. Compared with the fermentation substrate prepared by the control group using a simplified process and common raw materials, the contents of main anti-nutritional factors such as glucosinolates from rapeseed meal and trypsin inhibitors from soybean meal in the innovative fermentation substrate are greatly reduced. On the one hand, this benefits from the pretreatment and screening of raw materials in the composite substrate, and on the other hand, it also owes to the effective degradation and transformation of these harmful substances by specific strains (such as detoxifying Bacillus and Aspergillus niger) in the core fermentation flora during the multi-stage fermentation process. At the same time, the innovative fermentation substrate is enriched with higher levels of small molecule peptides and lactic acid, and the viable count of Lactobacillus plantarum, one of the main fermentation forces, is also significantly increased, which directly reflects the positive effects of the synergistic action of multiple strains and the refined fermentation process on the conversion of nutrients and the accumulation of beneficial metabolites.

[0123] Test Example 2: 1. Experimental Purpose This experiment aims to evaluate the protection effect and targeted release characteristics of the targeted slow-release microcapsule technology described in the present invention on core active ingredients (such as probiotics and plant extracts) under in vitro simulated digestion conditions, the retention rate and stability of key active ingredients in the finished feed, and verify its positive effects on specific physiological indexes of meat-type fighting cocks through a short-term feeding experiment. By comparing with the uncoated components and simplified coated components, the innovation and superiority of the microcapsule technology of the present invention are highlighted.

[0124] 2. Experimental Design and Materials Test Samples / Feeds: Sample Group 1 (M-EXP): Targeted slow-release microcapsules prepared according to the method of Example 1. Sample Group 2 (M-UNCOAT): A mixture of Core Component A and Core Component B of the targeted slow-release microcapsules in Example 1, without any coating treatment. Sample Group 3 (M-SIMP): Simplified coated microcapsules (single-layer ordinary coating) prepared according to the method of Comparative Example 3.

[0125] Feed Group 1 (F-EXP): Complete compound feed prepared according to the method of Example 1 (containing M-EXP).

[0126] Feed Group 2 (F-CON2): Compound feed prepared according to the method of Comparative Example 2 (the core active components are directly added in an uncoated form, that is, using M-UNCOAT instead of microcapsules, and other components and processes are the same as in Example 1).

[0127] Feed group 3 (F-CON3): Compound feed prepared according to the method of Comparative Example 3 (containing M-SIMP, and the final feed is granulated by high temperature).

[0128] To ensure comparability, the fermented base material in all feeds was prepared by the method of Example 1, and the proportions of other conventional feed components were the same. The differences were only in the treatment method and addition of the microcapsule / core component, and the granulation process of F-CON3. The addition amount of the core component (or its microcapsule form) was equivalent to the amount of the active substance contained in 0.3 parts by mass of the targeted sustained-release microcapsule in Example 1.

[0129] Experimental animals: Ninety healthy male meat fighting cocks at 22 days old (early brooding completed, similar body weights) were selected and randomly divided into 3 treatment groups (corresponding to feed groups F-EXP, F-CON2, F-CON3), with 6 replicates in each group and 5 chickens in each replicate.

[0130] Main reagents and instruments: Simulated gastric juice (SGF, containing pepsin), simulated intestinal juice (SIF, containing trypsin and bile salts), constant temperature shaking incubator, high performance liquid chromatograph (HPLC), gas chromatograph (GC), real-time fluorescence quantitative PCR instrument, microplate reader, centrifuge, pH meter, homogenizer, etc. Relevant standard products and reagent kits (such as icariin, astragalus polysaccharide, α-linolenic acid, conjugated linoleic acid, vitamin E, selenium, butyric acid standard products, probiotic culture medium, DNA extraction kit, etc.).

[0131] 3. Experimental procedures 3.1 Stability and release characteristics of core active ingredients under in vitro simulated digestion conditions Sample preparation: Accurately weigh equal amounts (based on the mass of the core active substance) of M-EXP, M-UNCOAT, and M-SIMP samples.

[0132] Simulated gastric digestion: Add the samples to conical flasks containing appropriate amounts of SGF respectively, and incubate at 39 °C (simulating the body temperature of poultry) with constant shaking (100 rpm) for 2 hours. Samples are taken at 0 hour and 2 hours respectively to measure the viable count of Clostridium butyricum in the samples (gradient dilution plate counting method) and the content of icariin (HPLC method, to evaluate whether it dissolves or degrades prematurely).

[0133] Simulated intestinal digestion: Centrifuge the sample after simulated gastric digestion (or inactivate pepsin by adjusting the pH), discard the supernatant (or directly adjust the pH and supplement the SIF components). Transfer the precipitate or all the digestive fluid to a conical flask containing an appropriate amount of SIF, and adjust the pH to 6.8 - 7.0. Continue to oscillate at a constant temperature of 39 °C (100 rpm). Sampling is carried out at 0.5 h, 1 h, 2 h, 4 h, and 6 h after the start of intestinal digestion. Determine the viable count of Clostridium butyricum released and the cumulative release rate of icariin in the sample (by HPLC method).

[0134] 3.2 Retention rate / stability of heat-sensitive and easily oxidized active ingredients in the finished feed Retention rate of heat-sensitive components (effect of pelleting): Respectively take the finished feed samples of F-EXP and F-CON3 after pelleting.

[0135] Determine the viable count of Clostridium butyricum in the feed (gradient dilution plate counting after grinding).

[0136] Determine the content of astragalus polysaccharide in the feed (such as by phenol-sulfuric acid method or HPLC method), compare it with the theoretical addition amount, and calculate the retention rate.

[0137] Stability of easily oxidized components (effect of storage): Respectively take the finished feed samples of F-EXP and F-CON2, and store them sealed under accelerated storage conditions of 37 °C and 75% relative humidity for 14 days.

[0138] Sampling is carried out on the 0th day and the 14th day. After extracting lipids, use GC method to determine the content changes of α-linolenic acid and conjugated linoleic acid, or use peroxide value (POV) or thiobarbituric acid reactive substances (TBARS) value to evaluate the degree of lipid oxidation.

[0139] 3.3 Effects of short-term feeding trials on specific physiological indicators Animal feeding: Feed 22-day-old meat fighting cocks with the corresponding F-EXP, F-CON2, and F-CON3 feeds according to the grouping, and the test period is 14 days. Other feeding management is the same as the conventional one.

[0140] Sample collection and index determination (at the end of the 14th day of the test): Randomly select 2 chickens from each replicate, collect blood from the heart, separate the serum, and store it at -20 °C.

[0141] [[ID=:34]]After sacrifice, aseptically collect cecal contents and store them at -80 °C.

[0142] Serum biochemical indicators: Use a kit or HPLC method to determine the concentration of natural vitamin E (α-tocopherol) in the serum.

[0143] The concentration of selenium in serum was determined by atomic absorption spectrometry or inductively coupled plasma mass spectrometry (ICP-MS).

[0144] Concentration of butyric acid in cecum: Cecal content samples were taken, and according to the method for determining VFA in Test Example 1, the concentration of butyric acid was mainly determined.

[0145] 4. Statistical analysis All data were analyzed using SPSS statistical software. One-way analysis of variance (ANOVA) and Duncan's multiple comparison test were used for between-group comparison. In vitro release data could be fitted to a release kinetic model. The results were expressed as mean ± standard deviation (Mean ± SD), and a P < 0.05 was considered to indicate a significant difference.

[0146] 5. Experimental data Table 2: Effects of different microcapsules / treatment methods on the in vitro release characteristics of core active ingredients, the stability of finished feed, and the physiological indexes of fighting cocks The results of the in vitro simulated digestion experiment strongly demonstrated the excellent protective ability and precise release characteristics of the targeted sustained-release microcapsules of the present invention for core active ingredients. Compared with the uncoated core components (M-UNCOAT) where a large amount of active ingredients (such as the viable count of Clostridium butyricum and icariin) were lost or prematurely dissolved in simulated gastric juice, the microcapsules with multi-layer composite coating (M-EXP) of the present invention significantly improved the stability of these components in an acidic environment, greatly increased the survival rate of Clostridium butyricum, and effectively controlled the premature dissolution of icariin. More importantly, under the subsequent simulated intestinal fluid conditions, M-EXP was able to achieve the effective release of core active ingredients, and both the cumulative released viable count of Clostridium butyricum and the cumulative release rate of icariin showed good performance. This was attributed to the pH-sensitive components (such as the combination of Eudragit® L100-55 and S100) in the outer coating material remaining intact in the gastric acid environment and starting to dissolve under the neutral pH conditions of the intestine, as well as the enzyme-sensitive components (such as substituted starch acetate) being further degraded under the action of specific intestinal enzymes, thus realizing the programmed delivery of active substances to specific parts of the intestine. In contrast, although the simply coated microcapsules (M-SIMP) had a certain protective effect, their protective effect in gastric juice and release control in intestinal fluid were both inferior to M-EXP.

[0147] Test Example 3: 1. Experimental purpose This experiment aims to comprehensively evaluate the comprehensive effects of the compound feed described in the present invention (based on Example 1) compared with several different control feeds (based on Comparative Examples 1, 2, and 3, representing simplified fermentation substrate, uncoated core components, simplified microcapsule coating, and high-temperature granulation, respectively) in terms of overall production performance, digestive tract health and function, body immunity and antioxidant status, and some biochemical indexes of muscle quality, so as to systematically verify the innovation and superiority of the overall technical solution of the present invention.

[0148] 2. Experimental design and animal grouping Experimental animals: 240 healthy 1-day-old male meat fighting cocks (of the same breed, with an initial average weight of about 42 ± 1.5 g) were selected and randomly divided into 4 treatment groups, with 6 replicates in each group and 15 chickens in each replicate.

[0149] Group 1 (EXP): Fed with the complete compound feed prepared based on Example 1.

[0150] Group 2 (CON1): Fed with the compound feed prepared based on Comparative Example 1 (simplified fermentation substrate, and the rest is the same as in Example 1).

[0151] Group 3 (CON2): Fed with the compound feed prepared based on Comparative Example 2 (the core active components are not coated, and the rest is the same as in Example 1).

[0152] Group 4 (CON3): Fed with the compound feed prepared based on Comparative Example 3 (simplified microcapsule coating and high-temperature granulation, and the rest is the same as in Example 1).

[0153] Feeding management: The experimental chickens were raised on the net under the same standardized environmental conditions (temperature, humidity, and light were regulated in stages according to the commercial broiler feeding guide), with free access to food and water. The experimental period was 42 days (6 weeks). The immunization program was carried out routinely.

[0154] Feed preparation: Feed for the EXP group: Prepared completely according to the method of Example 1.

[0155] Feed for the CON1 group: The fermentation substrate was prepared according to the method of Comparative Example 1, and the targeted sustained-release microcapsules were prepared according to the method of Example 1. The rest of the conventional components and the final feed preparation process (such as mixing, conditioning, and low-temperature granulation) were the same as in Example 1.

[0156] Feed for the CON2 group: The fermentation substrate was prepared according to the method of Example 1. The core active components A and B in the original microcapsules were directly mixed with other components in the manner of Comparative Example 2 (i.e., not coated). The rest of the conventional components and the final feed preparation process were the same as in Example 1.

[0157] CON3 group feed: The fermentation substrate was prepared according to the method of Example 1, and the microcapsules were prepared according to the method of Comparative Example 3 (simplified coating). The final feed was processed by the high-temperature granulation process described in Comparative Example 3.

[0158] In all feeds, the addition ratio of the fermentation substrate was 30% (by mass), and the total addition amount of the core active components (whether coated or not) was equivalent to the addition level of the microcapsules in Example 1. The types and ratios of other conventional feed components were kept consistent among groups (except for the differences in the treatment methods of the fermentation substrate and the core components).

[0159] 3. Experimental procedures and index determination 3.1 Comparison of production performance throughout the period Data recording: On the 1st, 21st, and 42nd days of the experiment, the chickens were weighed on an empty stomach in units of replicates.

[0160] Record the feed consumption of each replicate in each stage (1 - 21 days, 22 - 42 days, 1 - 42 days).

[0161] Observe and record the health status, death, and culling of the chicken flock daily.

[0162] Index calculation: Calculate the average daily gain (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR) by stage and throughout the whole process. Correct the feed consumption of dead and culled chickens.

[0163] 3.2 Assessment of digestive tract health and function (Day 42) Sample collection: At the end of the 42nd day of the experiment, randomly select 2 chickens close to the average body weight from each replicate. After sacrificing by carotid artery exsanguination, immediately collect the cecal contents aseptically. Part of it is used for pH value determination, and the rest is quickly stored at -80 °C for standby.

[0164] Determination of cecal content pH value: Conduct according to the method in Test Example 1.

[0165] Determination of volatile fatty acids (VFAs): According to the method in Test Example 1, determine the concentration of total volatile fatty acids (TVFA) and the molar percentages of acetic acid, propionic acid, and butyric acid.

[0166] 3.3 Assessment of body immunity and antioxidant status (Day 42) Sample collection: Before sacrifice, collect blood from the hearts of the selected chickens, separate the serum, and store it at -20 °C for standby. After sacrifice, quickly take liver tissue, freeze it in liquid nitrogen and store it at -80 °C.

[0167] Serum immune index: Use an ELISA kit to determine the concentration of immunoglobulin IgY (or IgG) in the serum.

[0168] Tissue antioxidant indicators: Take appropriate amount of liver tissue to prepare homogenate.

[0169] Superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) activity, and malondialdehyde (MDA) content in liver homogenates were determined using commercial kits. Results were expressed as enzyme activity units or content per milligram of tissue protein.

[0170] 3.4 Assessment of Muscle Quality-Related Biochemical Indices (Day 42, Optional) Sample collection: Left breast muscle samples were collected from the above-selected slaughtered chickens, and visible fat and connective tissue were removed. A portion was used for routine nutrient analysis, and a portion was vacuum-packed and stored at -80°C for fatty acid analysis.

[0171] Conventional nutritional ingredients: The crude protein and crude fat content in breast muscle were determined using the national standard method.

[0172] Specific fatty acid content: Total breast muscle lipids were extracted and methylated, and then the contents of α-linolenic acid and conjugated linoleic acid (CLA, the sum of major isomers) in the muscle were determined by gas chromatography-FID (GC-FID). The results were expressed as a percentage of the total fatty acids or as a percentage per 100 g of muscle.

[0173] 4. Statistical Analysis All data were analyzed using SPSS statistical software. One-way analysis of variance (ANOVA) was performed on the measurement data. If significant differences were found (P < 0.05), Duncan's or Tukey's HSD test was used for multiple comparisons. Results are expressed as mean ± standard deviation (mean ± SD).

[0174] 5. Experimental Data Table 3: Effects of different compound feeds on production performance, intestinal health, immune and antioxidant properties, and muscle quality of broiler fighting chickens during the entire period (1-42 days of age) Performance data throughout the entire production process showed that the experimental group (EXP) of broiler fighting chickens fed the complete compound feed of the present invention achieved significantly better average final weight and average daily weight gain throughout the entire process than the control groups fed a simplified fermentation base (CON1), uncoated core active ingredients (CON2), or a simplified coating and high-temperature pelleting (CON3). Feed conversion rates were also improved throughout the entire process. This demonstrates that the present invention significantly enhances the overall nutritional value and utilization efficiency of the feed by combining the optimized preparation of the fermentation base (as shown in Example 1, providing a more digestible, nutrient-rich matrix with lower anti-nutritional factor content) with the precise coating and protection of the core functional ingredients by targeted sustained-release microcapsules (as shown in Examples 1 and 2, ensuring stable and targeted release of the active ingredients). This synergistic effect enables animals to more efficiently convert ingested nutrients into weight gain, demonstrating the potential of the present invention's technical solution in improving the efficiency of intensive farming.

[0175] Improvements in digestive tract health, immunity, and antioxidant status further demonstrate the comprehensive advantages of the compound feed presented herein. The pH of the cecal contents of the experimental group (EXP) fighting chickens was closer to the optimal range, and the total volatile fatty acid (TVFA) content, particularly the molar percentage of butyric acid, was significantly higher than in the control groups. This directly reflects the positive regulatory effects of the present invention on the intestinal microbiome through the high-quality fermentation base and targeted delivery of probiotics (such as Clostridium butyricum), promoting the production of beneficial metabolites and thus maintaining intestinal barrier function and health. Furthermore, serum immunoglobulin IgY concentrations and liver superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the EXP group were significantly increased, while the content of the lipid peroxidation product malondialdehyde (MDA) was significantly reduced. This is attributed to the fact that the microencapsulation technology of the present invention effectively protects and delivers plant extracts and highly effective antioxidants (such as natural vitamin E and organic selenium) with immunomodulatory functions, enabling them to fully exert their effects in the animal body and enhance the body's nonspecific immunity and anti-oxidative stress ability. However, these effects were weakened in the control group due to poor quality of the fermentation base or insufficient protection and impaired activity of the core components.

[0176] 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. An efficient fattening compound feed for meat - using fighting cocks, characterized in that, By mass parts, it includes: 20 - 40 parts of fermentation base material; the fermentation base material is prepared by multi - stage solid - state symbiotic fermentation of a composite substrate by a core fermentation flora. The composite substrate contains 30 - 40% rapeseed meal, 20 - 30% pretreated fermented soybean meal, 15 - 25% corn germ meal, and 10 - 20% composite fruit and vegetable processing by - products by dry matter weight percentage; the core fermentation flora includes detoxifying Bacillus, Aspergillus niger, Lactobacillus plantarum, and Saccharomyces cerevisiae; 0.1 - 1.0 part of targeted slow - release microcapsule; the targeted slow - release microcapsule contains core component A and core component B, and adopts at least a double - layer coating structure. The coating structure contains a pH - sensitive enteric - coated material; core component A contains Clostridium butyricum and a plant extract complex. The plant extract complex contains Astragalus membranaceus extract, Epimedium brevicornum extract, and Atractylodes macrocephala extract; core component B contains a mixture of α - linolenic acid and conjugated linoleic acid, and a combination of natural vitamin E and organic selenium; Other conventional feed components, up to 100 parts. The other conventional feed components are selected from one or more of energy feed, protein feed, mineral feed, vitamin premix, and trace element premix.

2. The high-efficiency fattening compound feed for meat-use fighting cocks according to claim 1, wherein The composite fruit and vegetable processing by - products in the fermentation base material are a mixture of apple pomace, tomato pomace, and citrus peel in a weight ratio of 1:1:0.

5.

3. The efficient fattening compound feed for meat - using fighting cocks according to claim 1 or 2, characterized in that, The core fermentation flora of the fermentation base material specifically includes detoxifying Bacillus strain XJ - 5, Aspergillus niger strain FZ - 2, Lactobacillus plantarum strain DG - 8, and Saccharomyces cerevisiae strain YM - 3.

4. The efficient fattening compound feed for meat - using fighting cocks according to claim 1, characterized in that, In the plant extract complex in core component A of the targeted slow - release microcapsule, the weight ratio of astragalus polysaccharide in Astragalus membranaceus extract, icariin in Epimedium brevicornum extract, and atractylenolide II in Atractylodes macrocephala extract is 5:1:0.

5.

5. A high-efficiency fattening compound feed for meat fighting cocks and its preparation method according to claim 1, characterized in that, In core component B of the targeted slow - release microcapsule, the weight ratio of α - linolenic acid to conjugated linoleic acid is 2.5:1, and the ratio of natural vitamin E to selenium element in organic selenium is 100 IU:1 mg Se.

6. A high-efficiency fattening compound feed for meat fighting chickens and its preparation method according to claim 1, 4 or 5, characterized in that, The at least double - layer coating structure of the targeted slow - release microcapsule also contains an enzyme - sensitive material to achieve the programmed targeted release of core component A and core component B in the digestive tract of fighting cocks.

7. A method for preparing the high-efficiency fattening compound feed for meat-use fighting cocks according to any one of claims 1-6, characterized in that, It includes the following steps: S1. Prepare the fermentation base material: A composite substrate containing rapeseed meal, pretreated fermented soybean meal, corn germ meal, and composite fruit and vegetable processing by - products is subjected to multi - stage solid - state symbiotic fermentation with a core fermentation flora. The core fermentation flora includes detoxifying Bacillus, Aspergillus niger, Lactobacillus plantarum, and Saccharomyces cerevisiae; the multi - stage solid - state symbiotic fermentation includes a step of dynamically inoculating subsequent dominant strains based on intermediate metabolites or enzyme activity indexes during the fermentation process; S2. Prepare the targeted slow - release microcapsule: Core component A containing Clostridium butyricum and a plant extract complex, and core component B containing α - linolenic acid, conjugated linoleic acid, natural vitamin E, and organic selenium are subjected to at least double - layer coating treatment. The coating treatment includes using a pH - sensitive enteric - coated material; S3. Mix the fermentation base material obtained in step S1, the targeted slow-release microcapsules obtained in step S2, and other conventional feed components evenly to obtain the compound feed.

8. The preparation method of an efficient fattening compound feed for meat - using fighting cocks according to claim 7, characterized in that, In step S1 for preparing the fermentation base material, the multi-stage solid-state symbiotic fermentation specifically includes: S1-1. First-stage fermentation: Under aerobic conditions at a temperature of 28-30°C and a material humidity of 56-60%, inoculate Aspergillus niger and Bacillus detoxificans and ferment for 48-72 hours. S1-2. Second-stage fermentation: When the key enzyme activity or the concentration of characteristic hydrolysis products in the first-stage fermentation reaches a preset threshold, inoculate Lactobacillus plantarum and Saccharomyces cerevisiae, and ferment for 72-96 hours under microaerobic or anaerobic conditions at a temperature of 26-28°C and a material humidity of 51-55%.

9. The preparation method of a high-efficiency fattening compound feed for meat-use fighting cocks according to claim 7, characterized in that, In step S2 for preparing the targeted slow-release microcapsules, the at least double-layer coating treatment further includes coating with an enzyme-sensitive material, and the enzyme-sensitive material is selected from at least one of starch acetate with a specific degree of substitution or modified pectin; through the combined use of the pH-sensitive enteric coating material and the enzyme-sensitive material, the programmed targeted release of core component A and core component B in different sections of the fighting cock digestive tract is achieved.

10. The preparation method of a high-efficiency fattening compound feed for meat-use fighting cocks according to claim 7, 8 or 9, characterized in that, After step S3, there are also conditioning and low-temperature granulation steps; the conditioning step is to condition the evenly mixed material at 60-68°C under a steam pressure of 0.2-0.3 MPa for 60-90 seconds; the low-temperature granulation step is to granulate the conditioned material under the condition that the temperature of the granule leaving the die is ≤70-75°C.

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