Mountain bamboo partridge ecological breeding method suitable for altitude gradient

Abstract

The application discloses a mountainous Mei flower chicken ecological breeding method suitable for altitude gradient, and belongs to the technical field of poultry breeding. The method comprises the following steps: (1) feeding compound functional feed in stages: preparing feed for chick, young chicken and adult chicken stages, adding a compound functional additive composed of honeysuckle extract, wormwood leaf extract, fishy grass extract, garlicin and methylotrophic bacillus agent in the feed, and increasing the adding amount with the growth stage; (2) gradual release management according to altitude: indoor brooding and outdoor release are divided, the temperature is controlled according to the altitude in the brooding stage, a transition period is set before release, and the outdoor activity time is gradually prolonged, and zoning rotational grazing is adopted in the release stage. The application synergistically acts through the stage-by-stage nutrition regulation, the altitude adaptation and the gradual release, and significantly improves the growth performance, the immunity, the meat quality and the breeding survival rate of the mountainous Mei flower chicken.

Description

Technical Field

[0001] This application relates to the field of poultry farming technology, and in particular to an ecological farming method for mountain plum blossom chickens adapted to altitude gradients. Background Technology

[0002] The Mountain Plum Blossom Chicken is a superior native chicken breed, originating from the Qinling-Bashan Mountains. Through long-term natural selection, it has developed unique advantages adapted to the mountainous environment. This breed has firm, tender meat with a rich, mellow flavor. Its natural inosinic acid content is ≥2.5mg / g, muscle fat content is ≤3.2%, and protein content is ≥22%. It exhibits strong tolerance to roughage and significant disease resistance, commanding a market premium of 30%-50% compared to ordinary native chickens.

[0003] However, the existing mountain plum blossom chicken farming model has the following technical defects: (a) Poor growth performance Under the existing breeding model, the average net weight gain of a single mountain plum blossom chicken at 120-150 days of age is only 0.23±0.09kg, with a feed conversion ratio as high as 34.64%, resulting in low feed utilization and a long breeding cycle.

[0004] (ii) Weak immune system and gut function Under current breeding conditions, the spleen index of chicken flocks is only 0.25±0.12, indicating poor development of immune organs; the intestinal microbial community structure is unbalanced, with harmful bacteria enriched and beneficial bacteria insufficient, resulting in low nutrient absorption efficiency.

[0005] (iii) Frequent outbreaks of epidemics and substandard quality Under the existing farming practices, the incidence of common diseases such as coccidiosis and Newcastle disease is as high as 23%; the shear force of the pectoral muscle reaches 32.71±1.76N, and the meat is too tough; the drip loss rate is 2.41±0.38%, indicating poor water retention; and the redness value of the pectoral muscle is only 14.88±1.23, resulting in a dull meat color.

[0006] In summary, existing mountain plum blossom chicken farming techniques suffer from low growth efficiency, poor immunity and disease resistance, and substandard meat quality. Summary of the Invention

[0007] This application discloses an ecological breeding method for mountain plum blossom chickens adapted to altitude gradients, in order to solve the technical problems existing in related technologies, such as low growth performance, weak immune and intestinal function, frequent disease outbreaks, and substandard meat quality of mountain plum blossom chickens.

[0008] To solve the above problems, this application adopts the following technical solution: In a first aspect, embodiments of this application provide an ecological breeding method for mountain plum blossom chickens adapted to altitude gradients, comprising the following steps: (1) Feed compound functional feed in stages: Prepare stage feeds suitable for chickens, including at least the chick stage, pullet stage, and adult stage; The stage feed contains a compound functional additive, which is composed of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin and methyl nutrient Bacillus inoculum in a weight ratio of (60-80):(5-15):(5-15):(3-8):(3-8). The active ingredient of the methyltrophic Bacillus agent is methyltrophic Bacillus GBacillus-9 with the preservation number CGMCC No.13337; As chickens progress from chicks to young chickens and then to adult chickens, the amount of the compound functional additive added to the feed increases at each stage. (2) Gradual grazing management based on altitude gradients: The feeding and management of chickens are divided into two stages: indoor brooding and outdoor free-range. During the indoor brooding stage, the brooding temperature is controlled according to the altitude of the breeding area, with the brooding temperature in high-altitude areas being higher than that in low-altitude areas. Before entering the outdoor free-range stage, a transition period is set up, and the outdoor activity time is gradually increased each day to allow the chickens to gradually adapt to the woodland environment. After entering the outdoor grazing stage, grazing is carried out in a zoned rotational grazing manner, with each rotational grazing area grazing for a predetermined number of days before resting.

[0009] The methylotrophic Bacillus agent used in this application has the active ingredient of Bacillus methylotrophicus GBacillus-9 strain. This strain was isolated from the intestine of the striped bamboo shark (Chiloscyllium plagiosum) and was deposited at the China General Microbiological Culture Collection Center (CGMCC) on November 24, 2016, with accession number CGMCC No. 13337. The basic characteristics and identification results of this strain are detailed in the authorized Chinese invention patent (application number: 201710044718.4, patent title: A Bacillus GBacillus-9 strain with antibacterial effect and its isolation method and application).

[0010] The technical solutions adopted in the embodiments of this application can achieve the following beneficial effects: (1) The ecological breeding method for mountain plum blossom chickens adapted to altitude gradients provided in this application adopts a phased and incremental addition of compound functional additives (composed of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin and methyl nutrient Bacillus inoculant in a weight ratio of (60-80):(5-15):(5-15):(3-8):(3-8)) to meet the nutritional needs of chickens in the chick, pullet, and adult stages, so that each component in the compound functional additives can play a targeted regulatory role at different stages. At the same time, combined with altitude-specific temperature and humidity control (the brooding temperature in high-altitude areas is 2-3℃ higher than that in low-altitude areas) and gradual adaptation during the transition period, environmental stress is effectively reduced. Comparative experiments have verified that, using the method described in this application, the average final weight of mountain plum blossom chickens at 150 days of age can reach 2.35±0.71 kg, and the average net weight gain per chicken from 121 to 150 days of age is 0.50±0.04 kg, which is 117.4% higher than that of the conventional free-range model (0.23±0.09 kg). The feed conversion ratio can be reduced from 34.64% to 16.34%, a reduction of about 52.8%, which greatly improves feed utilization and significantly shortens the breeding cycle.

[0011] (2) The ecological breeding method for mountain plum blossom chickens adapted to altitude gradients provided in this application contains plant extracts (honeysuckle, mugwort, and houttuynia cordata) in the compound functional additives, which have anti-inflammatory and antibacterial effects. Allicin can enhance the activity of immune cells, and methyl-nutritive Bacillus can regulate the intestinal immune barrier. The four work synergistically to improve the immune level from within the body. At the same time, the transitional gradual adaptation and zoned rotational grazing and rest mechanism are adopted to reduce environmental stress on the chicken flock and reduce the risk of pathogen accumulation. According to the test, after adopting the method of this application, the spleen index of the chicken flock can be increased from 0.25±0.12 to 0.43±0.19, an increase of 72%; the serum IgM and IgG levels are significantly higher than those of the control group (P<0.05), and the IL-10 anti-inflammatory factor level is significantly increased; the disease incidence rate is reduced from 23% to below 3%, a decrease of 86.96%, and the survival rate is increased from 78% to over 95%, an increase of 21.79%.

[0012] (3) The ecological farming method for mountain plum blossom chickens adapted to altitude gradients provided in this application optimizes the meat quality of mountain plum blossom chickens in all aspects through rotational grazing management, the addition of compound grass powder to feed, and the phased and incremental addition of compound functional additives, combined with the ecological farming method based on altitude gradients. ① Tenderness and water retention: The shear force of the pectoral muscle can be reduced from 32.71N±1.76N to 23.63N±2.90N, a decrease of 27.8%, resulting in tender and juicy meat; the drip loss rate of the pectoral muscle can be reduced from 2.41%±0.38% to 1.70%±0.65%, a decrease of 29.5%, resulting in enhanced water retention and extended shelf life; ② Flesh color: The redness value (A value) of the pectoral muscle can be increased from 14.88±1.23 to 17.98±1.02, an increase of 20.8%, resulting in brighter flesh color and significantly improved product appearance; ③ Fatty acid composition: Saturated fatty acids: In pectoral muscle, caprylic acid (C8:0) showed an increasing trend with increasing additive dosage (EG2 and EG3 groups were significantly higher than the control group). In leg muscle, caprylic acid (C8:0), undecanoic acid (C11:0), palmitic acid (C16:0), and docosuccinic acid (C22:0) all showed an increasing trend with increasing dosage, with EG3 group being significantly higher than the control group. Monounsaturated fatty acids: The total amount of EG3 in the breast muscle group was 9.05% higher than that in the control group, and the total amount in all additive groups in the leg muscle was significantly higher than that in the control group; oleic acid (C18:1) and erucic acid (C22:1) were significantly increased in the additive groups, which helps to improve the tenderness and juiciness of the meat and cardiovascular health. Polyunsaturated fatty acids: The total amount of EG1 in the pectoral muscle group was not significantly different from that in the control group, while it was higher in both the EG2 and EG3 groups. The total amount of EG1 and EG2 in the leg muscle group was not significantly different from that in the control group, while it was higher in the EG3 group. Linoleic acid (C18:2) was higher in all three groups. ④ Market value: The market premium for high-quality commercial chickens has expanded from 30%-50% in the conventional model to 40%-60%. Attached Figure Description

[0013] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0014] Figure 1 A is a graph showing the serum urea nitrogen (BUN) content of each group of mountain plum blossom chickens; Figure 1 B is a graph showing the serum glucose (Glu) content of each group of mountain plum blossom chickens; Figure 1 C is a graph showing the detection results of serum aspartate aminotransferase (AST) activity in each group of mountain plum-blossom chickens; Figure 2 D is a graph showing the serum alanine aminotransferase (ALT) activity detection results of each group of mountain plum blossom chickens; Figure 2 E is a graph showing the results of serum total protein (TP) content detection in each group of mountain plum blossom chickens; Figure 2F is a graph showing the results of serum total cholesterol (TC) content detection in each group of mountain plum blossom chickens; Figure 3 G is a graph showing the serum triglyceride (TG) content of each group of mountain plum blossom chickens; Figure 3 H is a graph showing the results of serum tumor necrosis factor-α (TNF-α) content detection in each group of mountain plum blossom chickens; Figure 3 Figure I shows the results of serum interleukin-6 (IL-6) content detection in each group of mountain plum blossom chickens; Figure 4 J is a graph showing the results of serum interleukin-10 (IL-10) content detection in each group of mountain plum blossom chickens; Figure 4 K is a graph showing the results of serum immunoglobulin G (IgG) content detection in each group of mountain plum blossom chickens; Figure 4 L is a graph showing the results of serum immunoglobulin A (IgA) content detection in each group of mountain plum blossom chickens; Figure 5 M is a graph showing the detection results of serum immunoglobulin M (IgM) content in each group of mountain plum blossom chickens; Figure 5 N is a graph showing the results of principal component analysis (PCA) of serum biochemical and immune indicators and growth performance of each group of mountain plum blossom chickens; Figure 6 A is a comparison chart of the Chao1 index of gut microbiota in mountain plum blossom chickens in the EG1 group and the CG group; Figure 6 B is a comparison chart of the Shannon diversity index of gut microbiota in mountain plum blossom chickens in the EG1 group and the CG group; Figure 6 C is a comparison of the abundance-based coverage estimation index (ACE) of gut microbiota in mountain plum-blossom chickens in the EG1 and CG groups; Figure 7 D is a comparison chart of the gut microbiota phylogenetic diversity index (PD_whole_tree) of mountain plum-blossom chickens in the EG1 group and the CG group; Figure 7 E is a comparison chart of the Simpson diversity index of gut microbiota in mountain plum blossom chickens in the EG1 group and the CG group; Figure 8 This is a principal coordinate analysis (PCoA) plot of the gut microbiota of mountain plum blossom chickens in groups EG1 and CG based on Bray-Curtis dissimilarity distance; Figure 9 G is a stacked bar chart of relative abundance of gut microbiota at the phylum level, showing the community composition and relative abundance distribution of the EG1 group and the CG group at the bacterial phylum level; Figure 9 H is a stacked bar chart of relative abundance of gut microbiota at the genus level, showing the community composition and relative abundance distribution of the EG1 group and the CG group at the bacterial genus level; Figure 10 A is a differential genus-level species abundance analysis plot, showing the bacterial genera and their statistical results that show significant differences between the EG1 and CG groups; Figure 10 B is a differential species abundance analysis plot, showing the bacterial species with significant differences between the EG1 group and the CG group and their statistical results; Figure 11 This is the principal component analysis (PCA) score plot of the EG1 and CG groups of mountain plum-feathered chickens, showing the overall molecular spectrum distribution of the EG1 and CON groups (CG, control group); Figure 12 This is a partial least squares discriminant analysis (PLS-DA) score plot of mountain pheasants in groups EG1 and CG, showing the inter-group discrimination between samples in group EG1 and group CON. Figure 13 This is a heatmap of differential molecular abundance in mountain pheasants from the EG1 and CG groups, showing the clustering results of expression patterns of all differentially expressed molecules in the EG1 and CON group samples. Figure 14 This is a differential molecular volcano plot of mountain sika chickens in the EG1 and CG groups, showing the distribution of molecules that are significantly upregulated, downregulated, and have no significant differences in the two groups. Figure 15 This is a volcano plot variant-bar plot (also called Log2FC ordination plot) of differential metabolites between EG1 and CG group mountain plum blossom chickens. Figure 16 This is a differential abundance score plot (DAScorePlot) of metabolic pathway enrichment analysis between the EG1 group and the CG group of mountain plum-flower chickens. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions of this application will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of them. Based on the embodiments in this application, all other implementation methods obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0016] The inventive concept of this application lies in: addressing the differentiated nutritional needs and physiological characteristics of mountain plum blossom chickens at different growth stages (chicks, young chickens, and adult chickens), as well as the impact of different altitude gradients (low altitude, medium altitude, and high altitude) on the environmental stress of the flock, creatively combining staged incremental compound functional additive feeding with gradual free-range management based on altitude gradients. Specifically, on the one hand, a compound functional additive is formed by compounding honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin, and methyl-nutritive Bacillus inoculant in a fixed ratio, and the addition amount is gradually increased with the growth stage of the chickens to achieve multiple synergistic effects of anti-inflammatory and antibacterial properties, regulation of intestinal flora, enhanced immunity, and improved meat quality; on the other hand, the indoor brooding temperature is adjusted according to altitude (high altitude is higher than low altitude), and a three-level gradual adaptation process of "indoor brooding → semi-intensive feeding transition → full forest free-range" is set up, combined with a zoned rotational grazing and rest mechanism to minimize environmental stress. By combining the above-mentioned phased incremental nutritional regulation, altitude-specific environmental adaptation, and gradual free-range management, the technical problems of low growth performance, weak immune and intestinal function, frequent disease outbreaks, and substandard meat quality of mountain plum blossom chickens in existing technologies have been solved. The comprehensive effect of achieving a disease incidence rate of ≤3%, a survival rate of ≥95%, and a feed conversion ratio increase of ≥18% has been achieved.

[0017] The following is in conjunction with the appendix Figures 1 to 16 This application provides a detailed description of an ecological breeding method for mountain plum blossom chickens through specific embodiments and application scenarios.

[0018] This application provides a method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients, including the following steps: (1) Feed compound functional feed in stages: Prepare stage feeds suitable for chickens, including at least the chick stage, pullet stage, and adult stage; The stage feed contains a compound functional additive, which is composed of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin and methyl nutrient Bacillus inoculum in a weight ratio of (60-80):(5-15):(5-15):(3-8):(3-8). The active ingredient of the methyltrophic Bacillus agent is methyltrophic Bacillus GBacillus-9 with the preservation number CGMCC No.13337; As chickens progress from chicks to young chickens and then to adult chickens, the amount of the compound functional additive added to the feed increases at each stage. (2) Gradual grazing management based on altitude gradients: The feeding and management of chickens are divided into two stages: indoor brooding and outdoor free-range. During the indoor brooding stage, the brooding temperature is controlled according to the altitude of the breeding area, with the brooding temperature in high-altitude areas being higher than that in low-altitude areas. Before entering the outdoor free-range stage, a transition period is set up, and the outdoor activity time is gradually increased each day to allow the chickens to gradually adapt to the woodland environment. After entering the outdoor grazing stage, grazing is carried out in a zoned rotational grazing manner, with each rotational grazing area grazing for a predetermined number of days before resting.

[0019] In some embodiments, the stage feeds all contain a basal diet and compound functional additives; wherein, the amount of compound functional additives added to the chick stage feed is 0.3%-0.7% of the basal diet weight, the amount added to the pullet stage feed is 0.5%-1.0% of the basal diet weight, and the amount added to the adult chicken stage feed is 0.5%-1.5% of the basal diet weight. It is understood that the above ranges correspond to the optimal control ranges for the chick, pullet, and adult chicken stages, respectively.

[0020] In some embodiments, the weight ratio of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin, and methyltrophic Bacillus inoculant in the compound functional additive is 70:10:10:5:5. It is understood that honeysuckle extract, as the main ingredient, provides the core anti-inflammatory and antibacterial effects; artemisia and houttuynia cordata assist in enhancing the antibacterial effect; allicin activates immune cells; and Bacillus inoculant regulates the intestinal microecology. The components can be adjusted within the range of (60-80):(5-15):(5-15):(3-8):(3-8).

[0021] In some implementations, the chick stage is 0-40 days old, the juvenile stage is 41-120 days old, and the adult stage is from 121 days old to slaughter. It is understood that 0-40 days old is the critical period for immune organ development, 41-120 days old is the rapid growth period, and after 121 days old is the period for meat flavor development. This division is based on the growth and development curve of the mountain plum blossom chicken, and in actual farming, it can fluctuate by 5-10 days depending on the flock condition and seasonal factors.

[0022] In some implementations, the indoor brooding stage is from 0 to 40 days old, the transition period is from 41 to 60 days old, and the outdoor free-range stage is from 61 days old to market age. It is understood that indoor brooding (0-40 days old) ensures survival rate, the transition period (41-60 days old) allows for gradual environmental adaptation, and outdoor free-range (after 61 days old) improves meat quality. This division corresponds to the age range of the feed stage.

[0023] In some embodiments, the basal diet of the stage feed comprises, by weight percentage: 60%-68% corn, 20%-26% soybean meal, 3%-7% wheat bran, 1%-3% rice bran, 0.8%-1.2% dicalcium phosphate, 1.5%-2.5% limestone powder, and 2.5%-3.5% micronutrient premix, with the sum of the weight percentages of the above components being 100%. The micronutrient premix contains vitamin A, vitamin D, vitamin E, copper sulfate, and methionine. It is understood that this basal diet formula (corn, soybean meal, wheat bran, rice bran, etc.) provides basic energy and protein for mountain plum-flower chickens. The ranges of each component are set based on the cost of common raw materials and nutritional requirements, and the total of 100% ensures the integrity of the formula. The vitamins and micronutrients in the micronutrient premix meet the micronutrient requirements of chickens at different stages.

[0024] In some embodiments, the rice bran in the chick feed is replaced with fishmeal. It is understood that partially replacing rice bran with fishmeal during the chick stage is to enhance the supply of high-quality animal protein and meet the high amino acid requirements of the chicks' rapid growth and immune organ development.

[0025] In some embodiments, starting from 31 days of age, compound forage powder is added to the stage feed of chickens. The compound forage powder is composed of alfalfa powder and leafy grass powder in a mass ratio of (5-7):(3-5). In the feed for chicks aged 31-40 days, the amount of compound forage powder added is 0.6%-1.0% of the weight of the basal diet. In the feed for pullets and adult chickens, the amount of compound forage powder added is 1.2%-1.8% of the weight of the basal diet. It is understood that this application, by precisely controlling the amount of compound forage powder added in stages, avoids the impact of excessive crude fiber intake on digestion and absorption in the early stages, and can make full use of the active ingredients such as dietary fiber, flavonoids and polysaccharides in the compound forage powder in the middle and late stages of growth, so as to improve intestinal health and enhance immunity and stress resistance.

[0026] In some embodiments, the preparation process of the stage feed includes: mixing the basal diet according to the formula and preheating it to 55℃-65℃, then adding the compound functional additive according to the formula and mixing evenly (generally stirring at a uniform speed for 10-20 minutes), and then granulating it using a low-temperature steam conditioning granulation process at 65℃-75℃, resulting in a pellet diameter of 2.5mm-4.5mm and a pellet hardness of 2.0kgf-3.0kgf; For stage feeds without added compound forage powder, the pellets obtained from pelleting are the finished stage feed products; for stage feeds requiring the addition of compound forage powder, the compound forage powder is mixed in after the pellets have cooled, and the amount added is determined as follows: In the feed for chicks aged 31-40 days, the amount of compound forage powder added is 0.6%-1.0% of the basal diet weight; in the feed for pullets and adult chickens, the amount of compound forage powder added is 1.2%-1.8% of the basal diet weight. When adding compound forage powder, a step-by-step premixing method is adopted. First, the compound forage powder is mixed evenly with a small amount of cooled feed pellets, and then the mixture is expanded to all feed pellets in the batch until it is evenly mixed to ensure the uniformity of mixing and avoid uneven local concentration that may affect feeding. The preparation method of the compound forage powder includes: selecting alfalfa in its early flowering stage and vigorous leafy grass as raw materials, harvesting them on a sunny morning, retaining the tender stems and leaves, and removing aged lignified straw and impurities; spreading them out to air dry naturally for 4-6 hours to remove surface free moisture, and then transferring them to a cool, ventilated place to air dry until the moisture content is ≤12%, avoiding nutrient loss caused by high-temperature exposure; mixing the dried alfalfa and leafy grass in the specified proportion, and pulverizing them to 40-60 mesh using a low-temperature, low-speed pulverizer; sieving to remove impurities after pulverization, and storing them in a dry, light-proof, and sealed environment to prevent moisture absorption and mold growth. This application retains the active nutrients, flavonoids, polysaccharides, and dietary fiber components of the forage through a standardized low-temperature processing technology, replacing part of the concentrated feed, improving intestinal health, enhancing the body's stress resistance and immunity, and adapting to the needs of mountain plum blossom chicken farming. The compound forage powder is added using a "step-by-step premixing" method, effectively ensuring the uniformity of mixing and avoiding differences in feed intake or nutritional imbalance caused by uneven local concentrations. Alfalfa and leafy grasses were combined at a ratio of (5-7):(3-5), which takes into account the synergistic effect of protein, vitamins, polysaccharides, and flavonoids. After being air-dried and pulverized at low temperature, the activity of the raw materials was maintained and the storage stability was extended. The synergistic design of the above process and formula is fully adapted to the needs of the phased ecological farming of mountain plum blossom chickens, and provides a material basis for improving their meat flavor, immune function and overall health.

[0027] In some embodiments, during the indoor brooding stage: when the altitude of the breeding area is ≤1200 meters, the brooding temperature is gradually reduced from 33℃-37℃ to 23℃-27℃, with a daily temperature decrease of 0.2℃-0.3℃, and the humidity is controlled at 55%-65%; When the altitude of the breeding area is >1200 meters, the brooding temperature is gradually reduced from 35℃-39℃ to 25℃-29℃, with a daily temperature decrease of 0.2℃-0.3℃, and the humidity is controlled at 55-65%. It is understandable that the brooding temperature in high-altitude areas (>1200m) is generally 2℃ higher than in mid-to-low-altitude areas (≤1200m), with an initial temperature of 35-39℃ and an ending temperature of 25-29℃. This is because the ambient temperature is lower and the diurnal temperature range is larger at high altitudes, resulting in greater heat loss for the chicks, requiring a higher ambient temperature to compensate. A daily temperature decrease of 0.2-0.3℃ and a humidity of 55-65% are standard brooding parameters to ensure the gradual development and maturation of the chicks' feathers and thermoregulation system.

[0028] In some implementations, during the transition period: When the altitude of the breeding area is ≤1200 meters, the initial outdoor activity time is 2-4 hours, which is increased by 0.5-1.5 hours every day until the animals are free-range all day. When the altitude of the breeding area is >1200 meters, the activity area should only be opened when the daytime temperature is ≥12℃. The initial outdoor activity time should be 1.5-2.5 hours, increasing by 1 hour every 2-4 days until full-day free-range breeding is achieved. It is understandable that in low-to-medium altitude areas (≤1200m), the temperature is higher and more stable, and extending the daily activity time (starting with 2-4 hours, increasing by 0.5-1.5 hours each day) allows for rapid environmental adaptation. In high-altitude areas (>1200m), the temperature is lower and fluctuates greatly. The area should only be opened during the warmer daytime hours (e.g., noon) when the temperature is ≥12℃ (e.g., 12℃-18℃), with a shorter initial activity time (1.5-2.5 hours), increasing by 1 hour every 2-4 days, to avoid wind chill stress with a more conservative approach. It should be noted that outdoor temperatures vary with the seasons. For example, from April to June, the temperature difference between day and night is large, and in low-to-medium altitude areas, it is also important to avoid prematurely extending the activity time when the temperature is below 12℃. It is recommended to monitor outdoor temperature changes for three consecutive days before implementation to determine the safe starting point each day, rather than relying solely on altitude for mechanical operation. Furthermore, temperature changes should be continuously monitored throughout the entire breeding cycle to rationally select the breeding period.

[0029] In some implementations, the partitioned rotational grazing includes: When the altitude of the breeding area is ≤1200 meters, it is divided into multiple rotational grazing areas according to the standard of 6-10 mu / 100 chickens. Each rotational grazing area is free-range for 15-25 days and rested for 35-45 days. Every 80-120 chickens are equipped with a 4-6 square meter mobile chicken house. When the altitude of the breeding area is >1200 meters, multiple rotational grazing areas are planned according to the standard of 5-8 mu / 100 chickens. Each rotational grazing area is free-range for 25-35 days and rested for 45-60 days. Every 80-120 chickens are equipped with a 4-6 square meter mobile chicken house. Understandably, this application scientifically adjusts rotational grazing parameters based on differences in altitude gradients. The core reason is that in low-to-medium altitude areas (≤1200 meters), the temperature is higher and the rainfall and heat conditions are better, resulting in faster grass growth and stronger regeneration capabilities. Therefore, a shorter grazing period (15-25 days) is adopted to avoid overgrazing by chickens and damage to the grass growth points. At the same time, a medium rest period (35-45 days) is sufficient for the grass to recover its biomass and achieve sustainable grassland utilization. In contrast, in high-altitude areas (>1200 meters), the temperature is lower, the diurnal temperature range is larger, the grass growth period is shorter, and regeneration is slower. Therefore, it is necessary to appropriately extend the grazing period (25-35 days) to ensure that the chickens have sufficient time to graze on enough natural grass and insects to meet their nutritional needs. At the same time, extending the rest period (45-60 days) allows the grass sufficient time to recover and prevents grassland degradation and soil erosion caused by overgrazing. Furthermore, high-altitude areas have strong ultraviolet radiation and relatively low vegetation cover, so appropriately extending the rest period helps reduce the accumulation of pathogens and parasites in rotational grazing areas. Both altitude conditions are equipped with mobile chicken houses of 4-6 square meters / 80-120 chickens for easy relocation. This zoned rotational grazing scheme, while ensuring the health of the grassland ecosystem, provides mountain plum-flower chickens with ample activity space and natural feed sources, effectively reducing environmental stress and pathogen exposure risks. This lays a solid environmental foundation for further optimization of meat quality and health levels through compound forage and compound functional additives.

[0030] In some implementation methods, immunization measures are also included, specifically: subcutaneous injection of Marek's disease vaccine 0.15-0.25 ml / bird at 1 day old; nasal administration of Newcastle disease vaccine 0.04-0.06 ml / bird at 10 days old; oral administration of infectious bursal disease vaccine at a concentration of 0.08-0.12 g / L at 14 days old; and intramuscular injection of E. coli vaccine 0.25-0.35 ml / bird at 50 days old when the altitude of the breeding area is >1200 meters. It is understood that Marek's disease, Newcastle disease, and infectious bursal disease are essential basic immunizations for chicks, and the vaccination times and dosages at 1, 10, and 14 days old are industry standards. In high-altitude areas, due to the cold climate and open environment, the risk of E. coli infection is increased; therefore, an additional intramuscular injection of E. coli vaccine (0.25-0.35 ml / bird) is given at 50 days old. The above immunization program has been experimentally verified to effectively reduce the incidence of diseases to below 3%, without the stress burden caused by repeated immunization.

[0031] In some implementations, biosecurity measures are also included, specifically: during the outdoor free-range stage: every 20-40 days, for 2-4 consecutive days each time, chickens are given a honeysuckle decoction in their drinking water at a concentration of 0.3-0.7 g / L, with the water temperature controlled at 22-28℃; every 15 days, chickens are disinfected with 0.3% povidone-iodine; monthly deworming with thiabendazole; and when changing grazing areas, a 3-7 cm thick layer of quicklime is placed for disinfection by the chickens stepping on it. It is understood that the honeysuckle decoction in the drinking water (0.3-0.7 g / L, 22-28℃) utilizes the anti-inflammatory and antibacterial components of honeysuckle, administered monthly for 2-4 consecutive days to enhance herd immunity. The water temperature is controlled to ensure palatability and the activity of the effective components. The quicklime basin (3-7 cm thick) is placed at the entrance of the grazing area and replaced each time the area is changed, utilizing the strong alkalinity of the quicklime to kill pathogens on the soles of the shoes of personnel entering and exiting, thus cutting off the route of cross-infection. The above measures are low-cost and easy to operate, making them suitable for large-scale under-forest farming.

[0032] In some implementations, predator control measures are also included, specifically: hanging reflective strips or old CDs every 1.5-2.5 meters; erecting a 2.0-2.4 meter high nylon mesh fence with a mesh size of (4-6) cm × (4-6) cm around the perimeter of the breeding area; preparing a powder by mixing realgar and wood ash in a weight ratio of 1:(2-4), and spreading a 0.3-0.7 meter wide protective strip along the inner side of the fence, spreading it once a month, and respreading it after rain; it is understood that reflective strips / Old CDs (spaced 1.5-2.5 meters apart) reflect light to scare away predators like eagles; nylon mesh fencing (2.0-2.4 meters high, 4-6 cm mesh) physically isolates weasels, wild cats, and other mammals; a powder made of realgar and wood ash in a 1:2-4 ratio is sprinkled along the inside of the fence for 0.3-0.7 meters. The arsenic sulfide in realgar has snake-repelling properties, while wood ash provides an alkaline environment and enhances adhesion. This powder is applied monthly, with reapplication after rain to maintain its effectiveness. This triple protection covers birds, mammals, snakes, and rodents, ensuring the safety of free-range chickens.

[0033] In some implementation methods, human interference control measures are also included, specifically: setting up a 40-60 meter wide and 1.5-2.0 meter high wooden fence between the breeding area and the external road; fixing feeding times at 6:30-7:30 am and 5:30-6:30 pm daily, and cleaning and disinfecting feeding equipment once a week; it is understood that the wooden fence can effectively block vehicle noise and human movement from disturbing the flock, preventing panic and stress-induced reduced feed intake. Fixing feeding times at 6:30-7:30 am and 5:30-6:30 pm helps the flock establish a stable feeding rhythm, reducing frequent contact between humans and chickens due to irregular feeding times. Cleaning and disinfecting feeding equipment once a week prevents the spread of moldy feed and pathogens through the feed troughs.

[0034] Some implementations also include monitoring measures, specifically: installing high-definition surveillance cameras in the foraging areas and water sources of rotational grazing areas, and monitoring in real time via a mobile app. This means installing high-definition surveillance cameras in areas where chickens frequently move, such as foraging areas and water sources, and remotely monitoring the distribution, mental state, feeding and drinking habits of the chickens, as well as the presence of predators (such as eagles, snakes, and weasels) via a mobile app. This measure compensates for the lag and blind spots of manual inspections, and is particularly suitable for large areas of woodland and remote mountainous regions, allowing for the immediate detection of anomalies and the implementation of countermeasures to minimize losses.

[0035] In some implementations, when implementing rotational grazing, the chicken coop adopts a movable slatted structure with slatted floors. To adapt to mountainous environments with different altitude gradients and to balance warmth, ventilation, and manure collection, the movable chicken coop is specifically designed as follows: In low-altitude areas, the chicken coop has a shoulder height of 1.8m-2.0m and a ridge height of 3.2m-3.5m. The walls are covered with a double layer of insulation cotton and waterproof membrane to cope with the high temperature and humidity environment in summer. The roof is equipped with openable ventilation windows to ensure air circulation. In high-altitude areas, the chicken coop has a shoulder height of 2.0m-2.2m and a ridge height of 3.5m-3.8m. The insulation layer is thickened, and double-layered tempered glass is used for lighting to balance warmth and light requirements and to resist the problem of large temperature differences between day and night in high-altitude areas. All chicken houses feature slatted floors with gaps 1.5cm-2.0cm wide, made of rot-resistant pine wood. Removable manure collection troughs are installed underneath, with an incline of 5°-8° to facilitate natural manure collection and prevent manure accumulation that could contaminate the coop environment. Inside the coop, one feed trough and one waterer are provided for every 10 chickens, positioned at a height suitable for the feeding habits of mountain plum-feathered chickens. Multi-level perches (40cm-50cm off the ground) are also provided to reduce the time chickens spend on the ground, minimizing direct contact between manure and the chickens and reducing the risk of disease transmission at the source. The coop locations avoid low-lying, waterlogged areas in the mountains. Lower-altitude coops have additional drainage ditches, while higher-altitude coops have their floors raised 25-35cm to prevent rainwater backflow and ground dampness.

[0036] In some implementations, chicken manure treatment employs a slatted chicken coop fermentation method. Specifically, fresh chicken manure from the collection trough is promptly transferred to the fermentation area and evenly mixed with sawdust in a 7:3 ratio. The sawdust absorbs excess moisture from the chicken manure, adjusting the fermentation humidity to 55%-65% (it should clump together when squeezed but crumble easily when released). Then, 0.5-1% of EM (Effective Microorganisms) inoculant is added based on the total weight of the chicken manure. After thorough mixing, the mixture is piled into a fermentation heap 1.2-1.5m high and 2-3m wide, covered with plastic film for sealed fermentation. The fermentation period is 15-20 days, during which the heap is turned over every 5 days to ensure complete fermentation. The fermented chicken manure organic fertilizer is then directly used as fertilization in rotational grazing areas' fallow forests, achieving the resource utilization of chicken manure.

[0037] The aforementioned rotational grazing, combined with chicken manure treatment and chicken coop design, forms a virtuous ecological cycle of "chicken-manure-forest-chicken": the rotational grazing model allows the forest vegetation to fully recover and rest, and the fermented chicken manure organic fertilizer nourishes the forest, improving soil fertility. Monitoring shows that the organic matter content of the forest soil increased by 12%, and the use of chemical fertilizers decreased by more than 30%. At the same time, the scientific chicken coop design reduces the probability of disease outbreaks in the flock, and precise rotational grazing and feeding management reduce feed waste and disease losses, shorten the breeding cycle, and improve the quality of the mountain plum blossom chickens at market, achieving a dual improvement in ecological and economic benefits.

[0038] The chickens raised in this application are slaughtered at 140-160 days of age, with the following characteristics at slaughter: weight ≥ 1.49 kg, feed conversion ratio ≤ 19.43%, breast muscle shear force ≤ 26.26 N, leg muscle drip loss rate ≤ 2.09%; breast muscle drip loss rate ≤ 1.93%, breast muscle redness ≥ 16.79. For chickens weighing ≥ 1.5 kg, inosinic acid content ≥ 3.2 mg / g, feed conversion ratio ≤ 2.5:1, breast muscle shear force ≤ 24 N, drip loss rate ≤ 1.8%, and meat color score ≥ 5.4. These slaughter indicators are based on the experimental verification results of the methods described in this application. Among these criteria, a weight of ≥1.49kg ensures the marketable size and economic benefits of the chickens; a feed conversion ratio ≤19.43% represents high feed conversion efficiency; a breast muscle shear force ≤26.26N corresponds to tender meat and excellent taste; a leg muscle drip loss rate ≤2.09%; and a breast muscle drip loss rate ≤1.93%, indicating strong water retention and long shelf life; furthermore, the meat has a bright color and excellent commercial appearance. These indicators collectively constitute the quality standards for high-quality mountain plum blossom chickens at market.

[0039] To more clearly illustrate the technical solution of this application, the following detailed description is provided in conjunction with specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application.

[0040] This application uses the Qinba Mountain Plum Blossom Chicken as the experimental animal, and raises it in the forest at a mid-altitude (1200 meters) in the Qinba Mountain area.

[0041] I. Feed Instructions: 1. Trace element premix: Purchased from Dazhou Hefeng Biotechnology Co., Ltd., which is Hefeng laying hen compound feed 733.

[0042] 2. Compound Grass Meal: This is a mixture of alfalfa meal and leafy grass meal in a 6:4 mass ratio. Preparation Process: Select alfalfa in its early flowering stage and leafy grass in its vigorous growth stage as raw materials. Harvest on a sunny morning, retaining fresh, tender stems and leaves, and removing aged, lignified stalks and impurities. After harvesting, spread out to air dry naturally for 4-6 hours to remove surface moisture, then transfer to a cool, ventilated place to air dry until the moisture content is ≤12%, avoiding high-temperature exposure that could lead to nutrient loss. Mix the dried alfalfa and leafy grass according to the above ratio, and pulverize using a low-temperature, low-speed grinder, controlling the particle size to 40-60 mesh. After pulverizing, sieve to remove impurities and store in a dry, light-proof, sealed environment to prevent moisture absorption and mold growth, ensuring stable grass meal quality. When using, employ a step-by-step premixing method: first, mix the grass meal evenly with a small amount of basal diet, then expand the mixing to the complete diet, ensuring uniform mixing and avoiding uneven local concentrations that could affect foraging.

[0043] 3. Methyltrophic Bacillus inoculum: Take a bacterial suspension containing Bacillus GBacillus-9 strain with preservation number CGMCC No. 13337 and inoculate it according to the following volume ratio: 5 mL of GBacillus-9 bacterial suspension is inoculated into 500 mL of LB liquid medium for culture to obtain GBacillus-9 seed culture; then 500 mL of seed culture is inoculated into a fermenter containing 10 L of LB liquid medium for expansion culture to obtain GBacillus-9 fermentation broth; the obtained GBacillus-9 fermentation broth is centrifuged at 6000–10000 rpm for 10–20 minutes, the bacterial precipitate is collected, and washed 1–2 times with sterile physiological saline. The washed bacterial precipitate is then freeze-dried under vacuum to obtain a powdered inoculum. Viable cell count testing shows that the viable cell content of the obtained powder is greater than 2 × 10⁻⁶. 9 CFU / g.

[0044] 4. The experimental group used chick stage feed 1 (1-30 days old): It consisted of a basal diet and compound functional additives. The basal diet included the following by weight percentage: corn 64%, soybean meal 23%, wheat bran 5%, fish meal 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%. The compound functional additives were added at 0.5% of the total weight of the basal diet. The weight ratio of each component in the compound functional additives was honeysuckle extract: artemisia extract: houttuynia cordata extract: allicin: methyl nutrient Bacillus inoculant = 70:10:10:5:5. Feed preparation process: After mixing the basic diet, preheat it to 60°C, add compound functional additives and stir for 15 minutes, then granulate it by low-temperature steam at 70°C to obtain pelleted feed with a pellet diameter of 3-4 mm and a hardness of 2.5 kgf.

[0045] 5. The experimental group used feed two for chicks (31-40 days old): This feed consisted of a basal diet, a compound functional additive, and compound forage powder. The basal diet, by weight percentage, included: corn 64%, soybean meal 23%, wheat bran 5%, fish meal 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%. The compound functional additive was added at 0.5% of the total weight of the basal diet, with the weight ratio of its components being honeysuckle extract: artemisia extract: houttuynia cordata extract: allicin methyl nutrient Bacillus inoculant = 70:10:10:5:5. The compound forage powder was added at 0.8% of the total weight of the basal diet. Feed preparation process: (1) Core pellet preparation: Mix the components of the basic diet evenly, preheat to 60°C, add compound functional additives and stir for 15 minutes, and granulate by low temperature steam at 70°C to obtain core pellets with a pellet diameter of 3-4 mm and a hardness of 2.5 kgf. Cool and set aside. (2) Forage powder after mixing: Mix the compound forage powder with the above core pellets in a step-by-step premixing manner: First, mix the forage powder with a small amount of core pellets evenly, and then expand the mixing to the whole batch of feed to ensure the uniformity of mixing and avoid uneven local concentration affecting feeding.

[0046] 6. The experimental group used feed for young chickens, with compound functional additives and compound forage powder added to the basal diet. The basal diet, by weight percentage, included: corn 64%, soybean meal 23%, wheat bran 5%, rice bran 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%. The amount of the compound functional additive added is 0.8% of the total weight of the basic diet; the weight ratio of each component in the compound functional additive is honeysuckle extract: artemisia extract: houttuynia cordata extract: allicin: methyl nutrient Bacillus inoculant = 70:10:10:5:5. The amount of the compound forage powder added is 1.5% of the total weight of the basal diet; Feed preparation process: The same as the preparation process of the chick stage feed 2 (31-40 days old) used in the experimental group.

[0047] 7. The experimental group used adult chicken feed: compound functional additives and compound forage powder were added to the basal diet. The basal diet included the following by weight percentage: corn 64%, soybean meal 23%, wheat bran 5%, rice bran 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%. The amount of the compound functional additive added is set according to groups (the weight ratio of each component in the compound functional additive is honeysuckle extract: artemisia extract: houttuynia cordata extract: allicin: methyl nutrient Bacillus inoculant = 70:10:10:5:5): EG1 group: 0.5% of the total weight of the basal diet; EG2 group: 1.0% of the total weight of the basal diet; EG3 group: 1.5% of the total weight of the basal diet. The amount of the compound forage powder added is 1.5% of the total weight of the basal diet; Feed preparation process: The same as the preparation process of the chick stage feed 2 (31-40 days old) used in the experimental group.

[0048] 8. Control group chick feed: fed only the basal diet, which by weight percentage included: corn 64%, soybean meal 23%, wheat bran 5%, fish meal 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%. Feed preparation process: after mixing the basal diet, the pellets were pelleted with a diameter of 3-4 mm and a hardness of 2.5 kgf.

[0049] 9. Control group feed for young chickens: Only the basic diet was fed, which by weight percentage included: corn 64%, soybean meal 23%, wheat bran 5%, rice bran 2%, dicalcium phosphate 1%, limestone powder 2%, and trace element premix 3%; feed preparation process: the same as the control group feed for chicks.

[0050] 10. The control group used adult chicken feed: the same as the pullet feed used in the control group.

[0051] II. Specific breeding and management methods: 1. Venue preparation: At an altitude of 1200 meters in the Qinling-Bashan Mountains, 40 mu of woodland with open terrain, lush vegetation, and clean water sources were selected. It was divided into 5 equal rotational grazing areas according to the standard of 8 mu per area, with a 1-meter-wide isolation belt set between the rotational grazing areas.

[0052] Indoor brooding stage (1-40 days old) and transition period (41-60 days old): Mobile chicken houses are built in the rotational grazing area. The chickens are raised in mobile chicken houses, which can also be used for rotational grazing during the outdoor free-range stage. 5 square meters of mobile chicken house are provided for every 100 chickens. In addition, a semi-open activity area equipped with shade netting is set up next to the chicken house where the chickens are located during the transition period to help the chickens gradually adapt to the outdoor environment.

[0053] Outdoor free-range stage (61-150 days old): 5㎡ of mobile chicken house is provided for every 100 chickens for rotational grazing.

[0054] Mobile chicken coops: Featuring a slatted structure, these coops are well-suited to mountainous environments, combining warmth, ventilation, and manure collection. The specific design is as follows: the coop has a shoulder height of 2.0m and a ridge height of 3.5m. The walls are covered with a double layer of insulation cotton and a waterproof membrane to cope with the high temperature and humidity of summer. The roof has openable ventilation windows to ensure air circulation. All coops use slatted floors with 2.0cm gaps, made of anti-corrosion pine wood. A removable manure collection trough is installed underneath, with an 8° incline to facilitate natural manure collection and prevent manure accumulation and pollution of the coop environment. Inside the coop, one feed trough and one waterer are provided for every 10 chickens, positioned at a height suitable for the feeding habits of mountain plum-feathered chickens. Multi-level perches (45cm off the ground) are also provided to reduce the time chickens spend on the ground, minimizing direct contact between manure and chickens and reducing the risk of disease transmission at the source. The location of the chicken coop should avoid low-lying, waterlogged areas in mountainous regions, and drainage ditches should be added to prevent rainwater backflow and ground dampness. During the outdoor free-range stage, the mobile chicken coop should be moved to the next rotational grazing area as needed, according to the rotational grazing plan.

[0055] A 2.2-meter-high nylon fence with 5×5cm mesh was erected around the chicken coop to prevent animals from entering. Three high-definition surveillance cameras were installed in key locations such as the foraging area and water source to achieve full coverage monitoring of the breeding area.

[0056] Chicken manure treatment employs a slatted chicken coop fermentation method. The specific steps are as follows: Fresh chicken manure from the collection trough is promptly transferred to the fermentation area and evenly mixed with sawdust at a ratio of 7:3. The sawdust absorbs excess moisture from the chicken manure, adjusting the fermentation humidity to 55%-65% (it should clump together when squeezed but crumble easily when released). Then, 0.5%-1% of EM (Effective Microorganisms) inoculant (purchased from Henan Wobao Biotechnology Co., Ltd., mainly including denitrifying bacteria, photosynthetic bacteria, lactic acid bacteria, yeast, actinomycetes, and filamentous bacteria, with an effective live bacteria count of 50 billion / g) is added to the total chicken manure mass. After thorough mixing, the mixture is piled into a fermentation heap 1.2-1.5m high and 2-3m wide, covered with plastic film for sealed fermentation. The fermentation cycle is 15-20 days, during which the heap is turned over every 5 days to ensure complete fermentation. The fermented chicken manure organic fertilizer is directly used as fertilization in rotational grazing areas' fallow forests, realizing the resource utilization of chicken manure.

[0057] After the site preparation was completed, one-day-old chicks were introduced to begin the experiment. 400 healthy, uniformly weighted one-day-old mountain plum blossom chickens were randomly divided into two groups: an experimental group (300 chicks) and a control group (CG group, 100 chicks). Both groups were mixed-fed from 1 to 120 days of age.

[0058] Experimental group (300 birds): From day 1 to 120, the feed formula and additive dosage were the same, and all were managed according to the methods described in this application. All other breeding conditions (environment, grazing management, disease prevention and control, etc.) of the experimental group were completely consistent and uniformly managed.

[0059] Control group (CG group, 100 animals): From day 1 to 120, animals were managed uniformly according to the method described below.

[0060] 2. 1-40 days old: (1) Experimental group: All were managed in accordance with the following indoor brooding stage (in chicken house).

[0061] Temperature, humidity, and light: From 1 to 40 days old, the chicken house uses a combination of underfloor heating and heat lamps for dual temperature control. The initial temperature is 37℃, and the temperature is reduced by 0.25℃ every day until it reaches about 27℃ by 40 days old. The humidity is kept stable at 60%. Lighting duration: 21 hours / day for 1 to 15 days old, and reduced to 18 hours / day for 16 to 40 days old.

[0062] Feed: For chicks aged 1-30 days, the experimental group was fed with chick stage feed one (1-30 days old); for chicks aged 31-40 days, the experimental group was fed with chick stage feed two (31-40 days old).

[0063] Immunization: 0.2 ml / animal subcutaneously injected with Marek's disease vaccine at 1 day old; 0.05 ml / animal intranasally administered with Newcastle disease vaccine at 10 days old; 0.1 g / L of infectious bursal disease vaccine administered via drinking water at 14 days old.

[0064] By 40 days of age, a total of 294 chicks had survived in the experimental group, with a survival rate of 98%. The chicks were healthy and their immune organs were developing normally. At this point, the flock was ready to enter the transition period.

[0065] (2) Control group: Feed: Feed the control group with the same feed for chicks during the chick stage; Temperature, humidity, light, and immunization management were all the same as in the experimental group.

[0066] 3. 41-60 days old: (1) Experimental group: All animals were managed according to the following transitional breeding methods: A transitional free-range model was initiated at 41 days of age to allow the flock to gradually adapt to the woodland environment.

[0067] Temperature, humidity, and light: For chickens aged 41-60 days, the temperature in the chicken house should be controlled at 24-27℃ initially (at 41 days old), and then reduced by 1-2℃ every 3-5 days until it reaches 20-22℃ at 60 days old; humidity should be kept stable at 60%±5%. The lighting duration should be maintained at 16 hours / day.

[0068] Feed: The experimental group was fed with feed for young chickens, and their feeding, droppings and activity were observed throughout the process.

[0069] Outdoor activities: The semi-open activity area (equipped with shade netting) opens at 9:00 AM daily, and the fully open activity area (woodland) opens at 11:00 AM. Initially, outdoor activity time is 3 hours, increasing by 1 hour each day, gradually transitioning to 8 hours. If the outdoor temperature is below 12℃, outdoor activities will be suspended or opening hours shortened for the day.

[0070] By 60 days of age, after a 20-day transition period, the flock was in good spirits, eating normally, and had formed droppings. They had fully adapted to the outdoor environment. There were no new deaths or new cases in the experimental group, and the number of surviving chickens remained at 294.

[0071] (2) Control group: Feed: Feed the control group with the feed for young chickens.

[0072] Temperature, humidity, and light were managed in the same way as the experimental group, and no outdoor activities were conducted.

[0073] 4. 61-120 days old: (1) Experimental group: All young chickens were managed according to the following free-range rearing method: From 61 days old, the chickens enter the free-range stage of the young chickens. During the free-range stage, it is necessary to monitor the activities of the flock and the situation of natural enemies in real time through a mobile APP.

[0074] Feed: Continue using the feed for the pullet stage of the experimental group.

[0075] Free-range management: Rotational grazing is adopted, with each rotational grazing area changing every 20 days. Each rotational grazing area is used for 20 days of free-range grazing followed by 40 days of rest. Each 100 chickens is provided with a 5㎡ mobile chicken house, which is moved once a week to an area with vigorous vegetation growth (within the same rotational grazing area).

[0076] Biosecurity measures: Administer honeysuckle decoction in drinking water (0.5g / L, water temperature 22-28℃) once a month for 3 consecutive days; disinfect chickens with 0.3% povidone-iodine every 15 days; deworm with thiabendazole (10mg / kg body weight) once a month; and disinfect with 5cm of quicklime when changing grazing areas, with chickens stepping on the lime.

[0077] Predator control measures: Hang reflective strips every 2 meters; the outer nylon mesh fence is 2.2m high; spread powder with a mass ratio of realgar:wood ash = 1:3 along the inner side of the fence, with a width of 0.5m, once a month, and respread after rain.

[0078] Extreme weather response measures: During heavy rain or snow, chickens should be taken inside the chicken house to avoid rain and snow; when the temperature is high (>32℃), add 0.1% electrolyte to the drinking water, use trees (such as planted chestnut trees) to provide shade, and use cooling equipment to cool the chicken house.

[0079] Human interference control measures: A 50m wide and 1.8m high wooden fence is set up between the breeding area and the road; feeding is scheduled at 7:00 am and 6:00 pm, and feeding equipment is cleaned and disinfected once a week.

[0080] By 120 days of age, the average weight of each chicken in the experimental group was about 1.85-1.88 kg, and a total of 294 chickens survived. The surviving chickens were in good spirits and no major diseases occurred.

[0081] (2) Control group: Feed: Continue using the same feed as the control group for young chickens.

[0082] Free-range management: Grazing is carried out within one of the rotational grazing areas, without rotational grazing.

[0083] Biosafety measures, natural enemy control measures, extreme weather response measures, and human interference control measures were all the same as those for the experimental group aged 61-120 days.

[0084] By 120 days of age, the average weight of a single chicken in the control group was about 1.90 kg, and a total of 78 chickens survived. No major diseases occurred among the surviving chickens.

[0085] 5. 121-150 days old: At 121 days of age (starting from the adult stage): the 294 surviving chickens in the experimental group were randomly re-divided into 3 subgroups, as follows: Experimental Group 1 (EG1 group): 98 animals, further subdivided into 6 subgroups (some subgroups had 17 animals, and some had 16 animals). Experimental Group 2 (EG2 group): 98 animals, further subdivided into 6 subgroups (some subgroups had 17 animals, and some had 16 animals). Experimental Group 3 (EG3 group): 98 animals, further subdivided into 6 subgroups (some subgroups had 17 animals, and some had 16 animals). Control group (CG group): 78 animals survived, which were also subdivided into 6 subgroups (23 animals in each subgroup).

[0086] (1) Experimental group: All chickens were managed according to the following fattening method during the adult stage: They enter the fattening stage of adult chickens at 121 days of age and are raised to slaughter at 150 days of age.

[0087] Feed: Chickens were fed the experimental group's stage feed separately according to the aforementioned groups EG1, EG2, and EG3.

[0088] The management of free-range animals, biosecurity measures, natural enemy control measures, extreme weather response measures, and human interference control measures were all the same as those for the experimental group aged 61-120 days.

[0089] (2) Control group: Feed: Feed the control group adult chickens with the same feed.

[0090] Free-range management: Grazing continues in the same rotational grazing area, without rotational grazing.

[0091] Biosafety measures, natural enemy control measures, extreme weather response measures, and human interference control measures were all the same as those for the experimental group aged 61-120 days.

[0092] They are raised to 150 days old before being sold.

[0093] IV. Experimental Examples: The experimental period was from 121 to 150 days of age, with animals raised to market age of 150 days. Growth performance, immune organ indices, serum biochemical and immune indicators, gut microbiota structure, meat quality, and liver metabolome were measured. The results are as follows.

[0094] 1. Growth performance and immune organ indicators The experiment measured the initial total weight (IBW) and final total weight (FBW) of each group, calculated the average net weight gain (ANG) and feed conversion ratio (FCR) per bird, and measured the organ indices of the heart, liver, and spleen. 1.1 Experimental Methods: (1) Growth performance determination In the CG, EG1, EG2, and EG3 groups, 10 chickens were randomly selected from each group (1 or 2 chickens from each of the 6 subgroups within each group). Each chicken was weighed at the beginning (121 days of age) and the end (150 days of age), and the initial total weight (IBW) and final total weight (FBW) were recorded. The average net weight gain per chicken (ANG) was calculated. ANG = (FBW-IBW) / Number of chickens in the experiment; During the experiment, the daily feed intake of the selected 10 chickens was recorded. After the experiment, the total feed consumption was calculated, and the feed conversion ratio (F / G) was determined. Material weight ratio (F / G) = (Total weight gain / Total material consumption) × 100%; (2) Organ index measurement After the experiment, 10 chickens from the same batch were randomly selected from each group, weighed, and euthanized by exsanguination via the jugular vein. The heart, liver, and spleen were dissected and separated, rinsed with physiological saline, and dried with filter paper. The weight of each organ was then measured, and the organ index was calculated using the following formula: Organ index = Fresh organ weight (g) / Live body weight (kg).

[0095] 1.2 Experimental results are shown in Table 1.

[0096] Table 1. Growth performance and immune organ index of mountain plum blossom chickens in each group (n=10)

[0097] Note: Experimental data were analyzed using SPSS software using one-way ANOVA. Results are expressed as mean ± standard deviation. Different lowercase letters in the same column indicate significant differences (P < 0.05), while the same letter (including common letters, such as 'a' and 'ab') indicates no significant differences (P > 0.05). n = 10, indicating that 10 chickens were randomly selected from each group; initial weight and final weight are the total weight of the 10 chickens selected.

[0098] As shown in Table 1, there was no significant difference in the initial total weight among the groups (P>0.05), indicating that the experimental groups were balanced and highly comparable.

[0099] Compared with the CG group, the total weight at the end of the trial period was significantly higher in the EG1, EG2, and EG3 groups, the average net weight gain per bird was significantly increased, and the feed conversion ratio was significantly lower. Specifically, in the CG group, the average net weight gain per bird was only 0.23±0.09 kg during the 121–150 day period, with a feed conversion ratio of 34.64%, indicating low feed utilization and slow growth. In contrast, the EG1 group achieved an average net weight gain of 0.50±0.04 kg per bird during the 121–150 day period, with a feed conversion ratio decreasing to 16.34%, demonstrating significantly improved growth performance and feed conversion efficiency compared to the CG group.

[0100] Regarding immune organs, there were no significant differences in cardiac and liver indices among the groups (P>0.05), indicating that the technical solution proposed in this application has no adverse effects on the development of important organs and is safe to use. The spleen index of the EG1 and EG2 groups was significantly higher than that of the CG group (P<0.05), indicating more complete development of immune organs and significantly enhanced immune function and disease resistance.

[0101] In summary, this application utilizes rotational grazing management, the addition of compound forage powder to feed, and the phased and incremental addition of compound functional additives, along with various technical means such as altitude-based transitional breeding, environmental buffering, and biosecurity control, to significantly improve the final weight and net weight gain of mountain plum blossom chickens, significantly reduce the feed conversion ratio, and increase the spleen index, thereby achieving synergistic optimization of growth performance and immune level.

[0102] 2. Serum biochemical and immune indicators To assess the impact of compound functional additives on the health status of mountain plum blossom chickens, serum biochemical and immunological parameters were measured.

[0103] 2.1 Experimental Methods: After the experiment, six chickens were randomly selected from each of the CG, EG1, EG2 and EG3 groups (one chicken from each of the six subgroups of each group) to collect blood from their jugular veins, and the serum was separated and stored at -20℃ for testing.

[0104] All biochemical and immunological indicators were detected using enzyme-linked immunosorbent assay (ELISA). All kits were purchased from Shandong Jiuyi Biotechnology Co., Ltd., and the operation was strictly carried out in accordance with the kit instructions.

[0105] Principal component analysis (PCA) was used to perform dimensionality reduction analysis on serum biochemical and growth parameters.

[0106] 2.2 Experimental Results: like Figures 1 to 5 As shown: (1) There were no significant differences in serum urea nitrogen (BUN), glucose (Glu), aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), and triglycerides (TG) among the groups (P>0.05), indicating that the technical solution of this application has no adverse effects on the liver and kidney function and glucose and lipid metabolism of mountain plum blossom chickens, and its use is safe.

[0107] (2) The total protein (TP) in the EG3 group was significantly higher than that in the CG group (P<0.05), indicating an improvement in the body's protein synthesis and nutrient metabolism capabilities. This suggests that the technical solution proposed in this application can improve the protein synthesis capabilities and nutrient metabolism levels of mountain plum blossom chickens.

[0108] (3) Regarding immune and inflammatory markers: The serum tumor necrosis factor-α (TNF-α) levels in both EG1 and EG2 groups were significantly lower than those in the CG group (P<0.01), and the serum interleukin-6 (IL-6) level in the EG1 group was significantly lower than that in the CG group (P<0.05), suggesting that this application can effectively alleviate the body's inflammatory response. Among humoral immune markers, the serum immunoglobulin G (IgG) level in the EG2 group was significantly higher than that in the CG group (P<0.05), and the serum immunoglobulin M (IgM) levels in the EG1 and EG3 groups were significantly (P<0.05) and extremely significantly (P<0.01) higher than those in the CG group, respectively, with the EG3 group showing the best IgM enhancement effect. There were no significant differences in serum immunoglobulin A (IgA) and interleukin-10 (IL-10) levels among the groups (P>0.05). The above results indicate that the compound functional additives of this application can effectively enhance the humoral immune function of mountain plum blossom chickens and have significant anti-inflammatory effects, with some indicators showing a dose-dependent enhancement trend.

[0109] (4) Principal component analysis (PCA) results showed that, based on serum biochemistry, immune indicators and growth performance, there was a clear clustering trend between the treatment groups and the control group. Among them, the EG1 group and the CG group had the most significant differentiation, indicating that the compound functional additive treatment of this application had a systematic effect on the overall physiological state of mountain plum blossom chickens, which is consistent with the aforementioned improvement results in growth performance, immune function and inflammatory indicators.

[0110] In summary, this application has achieved systematic physiological optimization in mountain plum-feed chickens through rotational grazing management, the addition of compound forage powder to feed, and the phased and incremental addition of compound functional additives, combined with standardized immunization programs, biosafety disinfection, physical isolation and protection, and environmental stress buffering. This has resulted in improved protein synthesis capacity, enhanced humoral immunity and anti-inflammatory effects without adverse effects on the liver, kidneys and metabolism. Principal component analysis results also support this conclusion.

[0111] 3. Gut microbial community structure 3.1 Experimental Methods After the experiment, six chickens were randomly selected from each of the CG and EG1 groups (one chicken from each of the six subgroups of each group). Fresh feces or cecal contents were aseptically collected, placed in cryovials, flash-frozen in liquid nitrogen, and stored at -80°C for later use.

[0112] Whole-genome DNA was extracted from the intestinal contents (rectum) of mountain-grown plum-flowered chickens using the CTAB method. DNA purity and concentration were determined by 1% agarose gel electrophoresis. The DNA was diluted to 1 ng / µL with sterile water, and PCR amplification of the V3-V4 region of the 16S rRNA gene was performed using primers 341F (5′-CCTAYGGGRBGCASCAG-3′) and 806R (5′-GGACTACNNGGGTATCTAAT-3′) designed by Wuhan Maiwei Metabolism Technology Co., Ltd. (Wuhan, China). A 16S rRNA gene amplicon library was constructed according to the Illumina sequencing standard protocol, and sequencing was performed by Wuhan Maiwei Metabolism Technology Co., Ltd. using the Illumina NovaSeq 6000 high-throughput sequencing platform.

[0113] To obtain high-quality sequence data, the raw sequencing reads were first spliced ​​and filtered. Then, the Deblur algorithm was applied for sequence denoising to generate amplicon variants (ASVs). Representative ASV sequences were taxonomically annotated using Mothur software (v1.48). The composition and structure of the microbial community were analyzed at both the phylum and genus levels. To identify common and unique ASVs among different samples and to assess species richness and evenness within samples, the α-diversity index was calculated. Based on the unweighted UniFrac distance matrix of ASV abundance, non-metric multidimensional scaling (NMDS) was used for β-diversity analysis to assess intergroup differences in gut microbial community structure under different feeding patterns.

[0114] 3.2 Experimental Results Alpha diversity: The Chao1 index and ACE index of the EG1 group were significantly lower than those of the CG group (P < 0.05), while the Shannon index, phylogenetic diversity (PD), and Simpson index showed no significant differences (P > 0.05), indicating that the EG1 group had a more streamlined and stable microbial community structure. Please refer to [link to relevant documentation]. Figure 6 A, Figure 6 B. Figure 6 C Figure 7 D、 Figure 7 E.

[0115] β-diversity: PCoA analysis based on Bray-Curtis distance showed significant separation between the EG1 and CG groups, with a significant difference in β-diversity between the groups (P < 0.05), indicating that the compound functional additive had a significant impact on the overall structure of the gut microbiota. Please refer to [link to relevant documentation]. Figure 8 .

[0116] At the phylum level: the dominant taxa in both groups EG1 and CG were Firmicutes, Proteobacteria, and Actinobacteria, followed by Cyanobacteria, Bacteroidetes, and Campylobacteria. No statistically significant differences were observed in the relative abundance of the top 10 most abundant phyla between the two groups; the composition of each group was similar. Please refer to [link to relevant documentation]. Figure 9 G.

[0117] At the genus level: the major genera in groups EG1 and CG include *Lactobacillus*, *Romobrulla*, and *Lactobacillus*, while *Bacillus*, *Corynebacterium*, and *Lactobacillus* are minor contributors. (See also...) Figure 9 H. Similarly, there were no significant differences in the relative abundance of the top 10 most abundant genera between the two groups. Please refer to [link to relevant documentation]. Figure 10 A.

[0118] At the species level: The abundance of *Citrus maxima* was significantly increased in group EG1; the abundance of *Lactobacillus reuteri*, *Corynebacterium diphtheriae*, and *Lactobacillaceae bacterium 28-4* was significantly increased in group CG. Please refer to [link to relevant documentation]. Figure 10 B.

[0119] In summary, this application significantly optimized the intestinal microecology of mountain plum-feathered chickens through rotational grazing management, the addition of compound forage powder to feed, and the phased, incremental addition of compound functional additives, combined with phased feeding model adaptation, environmental disinfection to reduce pathogen accumulation, and a low-stress environment to stabilize the intestinal microecology. Specifically, the Chao1 and ACE indices of the EG1 group were significantly reduced, and the microbiota structure became more streamlined and stable; β-diversity analysis showed a clear separation between the EG1 group and the control group, with a significant change in the overall structure of the intestinal microbiota; at the species level, the abundance of Citrus species was significantly increased in the EG1 group, while the abundance of conditional pathogens such as Lactobacillus reuteri and Corynebacterium diphtheriae was significantly increased in the CG group. These results indicate that the technical solution of this application can target and regulate the intestinal microbiota, reduce potential pathogens, enhance the competitive advantage of beneficial bacteria, and thus establish a healthier intestinal microecological environment.

[0120] 4. Meat quality at 150 days old 4.1 Experimental Methods After the experiment, six chickens were randomly selected from each of the CG and EG1 groups (one chicken from each of the six subgroups of each group). After euthanasia by exjugation through the jugular vein, samples of the pectoral muscle (pectoralis major) and leg muscle (thigh muscle) were quickly collected.

[0121] Referring to NY / T 1333-2007 "Determination of Livestock and Poultry Meat Quality", a colorimeter was used to determine meat color (brightness L, redness A, yellowness B), a pH meter was used to determine the muscle pH value at 1h, 24h, and 48h after slaughter, a shear force meter was used to determine muscle tenderness (shear force value, N), and a pressure method was used to determine muscle water-holding capacity (drip water loss rate, cooking loss rate).

[0122] Referring to the national food safety standard (GB5009.168-2016), the fatty acid composition of muscle tissue was determined using a gas chromatograph (7890B, Agilent Technologies, USA), and the compounds were quantitatively analyzed by the relative content of peak areas of each component.

[0123] 4.2 Experimental Results As shown in Tables 2 to 8 below.

[0124] Table 2 Quality indicators of mountain plum blossom chicken meat in each group (n=6)

[0125] Note: The experimental data were analyzed by one-way ANOVA using SPSS software. The results are expressed as mean ± standard deviation. P < 0.05 indicates a significant difference. Different lowercase letters in the same column indicate significant differences (P < 0.05), while the same letter (including common letters, such as a and ab) indicates no significant difference (P > 0.05).

[0126] n=6 means that 6 experimental chickens are randomly selected from each group.

[0127] As shown in Table 2: Compared with the CG group, the cooking loss rate of the EG1, EG2, and EG3 groups was slightly reduced, the drip loss rate of the breast muscle was significantly reduced (P < 0.05), and there was no significant difference in the drip loss rate of the leg muscle (P > 0.05), indicating a significant improvement in muscle water retention. There was no significant difference in pH value between the breast and leg muscles. The redness A value of the breast muscle was significantly increased (P < 0.05), and the yellowness B value of the leg muscle was significantly decreased (P < 0.05), resulting in bright and uniform meat color. The shear force of the breast muscle decreased from 32.71 ± 1.76 N in the CG group to 23.63 ± 2.90 N in the EG3 group (P < 0.05), significantly improving meat tenderness and comprehensively optimizing flavor and commercial appearance. This demonstrates that this application significantly improved the muscle quality of mountain plum blossom chickens through rotational grazing management, the addition of compound forage powder to the feed, and the phased, incremental addition of compound functional additives.

[0128] Table 3. Results of saturated fatty acid detection in the breast muscle of mountain plum blossom chickens in each group (n=6)

[0129] Table 4. Results of monounsaturated fatty acid detection in the breast muscle of mountain plum blossom chickens in each group (n=6)

[0130] Table 5. Results of polyunsaturated fatty acid detection in the breast muscles of mountain plum blossom chickens in each group (n=6)

[0131] In Tables 3 to 5, the experimental data were analyzed using one-way ANOVA with SPSS software. The results are expressed as mean ± standard deviation. Different lowercase letters in the same column indicate significant differences (P < 0.05), while the same letter (including common letters, such as a and ab) indicates no significant differences (P > 0.05). n=6 indicates that 6 experimental chickens were randomly selected from each group.

[0132] From Tables 3 to 5, we can see that: A total of 12 fatty acids were detected in the pectoral muscle samples, covering three major categories: saturated fatty acids (5 types), monounsaturated fatty acids (5 types), and polyunsaturated fatty acids (2 types).

[0133] It should be noted that the control group was fed conventional stall feeding and basal diet without the addition of compound grass powder and compound functional additives, and no rotational grazing was carried out; while the experimental groups (EG1, EG2, EG3) under the rotational grazing conditions had compound grass powder added to the feed in proportion, and compound functional additives were added in stages.

[0134] ①Saturated fatty acids: The EG2 group had the highest saturated fatty acid content, which was significantly higher than that of the CG group (P<0.05). The C8:0 ratio increased with increasing dosage (the EG2 and EG3 groups were significantly higher than the CG group, while the C22:0 ratio decreased. The CG group was significantly higher than the EG1 and EG2 groups). ② Monounsaturated fatty acids: The content of EG3 group was significantly higher than that of CG group and EG1 group (P<0.05), and the contents of C15:1, C22:1 and C24:1 were significantly increased in EG2 and EG3 groups (P<0.05). ③ Polyunsaturated fatty acids: There was no significant difference in the total amount of polyunsaturated fatty acids in the pectoral muscles among the groups (P>0.05), with the EG2 and EG3 groups being slightly higher than the CG group.

[0135] In summary, this application significantly optimized the fatty acid composition of the breast muscle of mountain-raised plum blossom chickens through rotational grazing management, the addition of compound forage meal to the feed, and the phased, incremental addition of compound functional additives. Specifically, the content of some medium- and short-chain saturated fatty acids (e.g., C8:0) was significantly increased, while the content of long-chain saturated fatty acids (C22:0) decreased. Simultaneously, the content of monounsaturated fatty acids was significantly increased, resulting in improved meat flavor and nutritional value without negatively impacting polyunsaturated fatty acids. These results further validate the significant advantages of the method presented in this application in improving chicken quality.

[0136] Table 6. Results of saturated fatty acid detection in the leg muscles of mountain plum blossom chickens in each group (n=6)

[0137] Table 7 Results of monounsaturated fatty acid detection in leg muscles of mountain plum blossom chickens in each group (n=6)

[0138] Table 8. Results of polyunsaturated fatty acid detection in the leg muscles of mountain plum blossom chickens in each group (n=6)

[0139] In Tables 6 to 8, the experimental data were analyzed using one-way ANOVA with SPSS software. The results are expressed as mean ± standard deviation. Different lowercase letters in the same column indicate significant differences (P < 0.05), while the same letter (including common letters, such as a and ab) indicates no significant differences (P > 0.05). n=6 indicates that 6 experimental chickens were randomly selected from each group.

[0140] From Tables 6 to 8, we can see that: A total of 14 fatty acids were detected in the leg muscle samples, covering three major categories: saturated fatty acids (7 types), monounsaturated fatty acids (5 types), and polyunsaturated fatty acids (2 types).

[0141] ①Saturated fatty acids: The EG3 group had the highest saturated fatty acid content (226.79 μg / g), which was significantly higher than other groups (P<0.05). The C8:0, C11:0, C16:0, and C22:0 ratios increased with the increase of the amount of compound functional additives. ② Monounsaturated fatty acids: The total amount of monounsaturated fatty acids in the EG1, EG2 and EG3 groups was significantly higher than that in the CG group (P<0.05), and the contents of C18:1 and C22:1 were significantly increased in the EG1, EG2 and EG3 groups (P<0.05). ③ Polyunsaturated fatty acids: The total amount of polyunsaturated fatty acids in the EG3 group was higher than that in other groups.

[0142] In summary, this application significantly optimized the fatty acid composition of mountain plum blossom chicken leg muscles through rotational grazing management, the addition of compound forage meal to the feed, and the phased, incremental addition of compound functional additives. The total saturated fatty acid content (especially C8:0, C11:0, C16:0, and C22:0) and the total monounsaturated fatty acid content (especially C18:1 and C22:1) were significantly increased, contributing to improved meat flavor and nutritional value. Although the DHA content decreased, the total polyunsaturated fatty acids were not significantly adversely affected, and the linoleic acid content increased. These results further validate the significant advantages of the method of this invention in improving chicken quality (including breast and leg muscles).

[0143] 5. Liver Metabolome 5.1 Experimental Methods: After the experiment, six chickens were randomly selected from each of the CG and EG1 groups (one chicken from each of the six subgroups of each group). After euthanizing the chickens by exjugation through the jugular vein, liver tissue was quickly collected, placed in cryovials, flash-frozen in liquid nitrogen, and stored at -80°C for later use.

[0144] 20 mg of liver tissue was collected, and metabolites were extracted using 70% methanol aqueous solution. After centrifugation, the supernatant was collected. Gradient separation was performed using a Waters HSS T3 column with 0.1% formic acid aqueous solution and acetonitrile as the mobile phase, and the results were detected by liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0145] The raw detection data were processed using XCMS software and compared with a metabolite database for identification. Metabolites with a score > 0.5 and a quality control coefficient of variation (QC-CV) < 0.3 were selected and retained. The positive and negative ion mode data were merged to obtain the final metabolite data matrix (all_sample_data.xlsx).

[0146] 5.2 Experimental Results PCA and PLS-DA analyses showed significant separation of metabolite clusters between the experimental and CG groups, indicating significant differences in liver metabolic profiles. (Please refer to [link / reference]). Figures 11 to 14 .

[0147] Compared to the CG group, the experimental group screened out 87 differentially regulated metabolites, of which 56 were upregulated and 31 were downregulated. To further understand the changes in differentially regulated metabolites, the top 20 metabolites with the highest fold changes were analyzed. This revealed that 17 metabolites were significantly upregulated and 3 were significantly downregulated. The upregulated metabolites mainly included: alph-tocopherolquinone, alph-tocopherol, phosphatidylserine (18:1–16:0), diglyceride (18:2–16:1–0:0), and phosphatidylserine (14:0–20:1). The downregulated metabolites mainly included: dihydroergotamine, triamterene, and delta-hexanolactone. Please refer to [link to relevant documentation]. Figure 15 ; KEGG pathway enrichment analysis showed that differentially metabolites were mainly enriched in the glycerophospholipid metabolism pathway, followed by significant enrichment in sulfur metabolism, cysteine ​​and methionine metabolism, pyrimidine metabolism, and bile secretion pathways. (See also...) Figure 16 .

[0148] In summary, this application significantly improved the liver metabolic profile of mountain plum-feathered chickens through rotational grazing management, the addition of compound forage powder to the feed, and the phased, incremental addition of compound functional additives, combined with altitude-specific environmental adaptation, human interference control, and improvement of the chickens' overall health. PCA and PLS-DA analyses showed that the metabolites in the experimental and control groups were clearly separated, with a total of 87 differentially expressed metabolites identified (56 upregulated and 31 downregulated). Among the top 20 metabolites with the largest fold changes, 17 were significantly upregulated (such as antioxidant-related metabolites like alph-tocopherolquinone, alph-tocopherol, and phosphatidylserine (PS)) and 3 were significantly downregulated. KEGG pathway enrichment analysis indicated that the differentially expressed metabolites were mainly enriched in pathways such as glycerophospholipid metabolism, sulfur metabolism, cysteine ​​metabolism, and methionine metabolism. The above results indicate that the technical solution of this application, through the synergistic effect of multiple links such as nutritional regulation, ecological free-range and liver metabolism optimization, not only improves muscle metabolism, meat flavor and reduces stress, but also further enhances the body's antioxidant capacity, thus achieving comprehensive optimization of the overall quality of mountain plum blossom chicken.

[0149] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0150] Furthermore, it should be noted that the scope of the methods and apparatus in the embodiments of this application is not limited to performing functions in the order shown or discussed, but may also include performing functions substantially simultaneously or in the reverse order, depending on the functions involved. For example, the described methods may be performed in a different order than described, and various steps may be added, omitted, or combined. In addition, features described with reference to certain examples may be combined in other examples.

[0151] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application.

Claims

1. A method for ecologically raising mountain plum blossom chickens adapted to altitude gradients, characterized in that, Includes the following steps: (1) Feed compound functional feed in stages: Prepare stage feeds suitable for chickens, including at least the chick stage, pullet stage, and adult stage; The stage feed contains a compound functional additive, which is composed of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin and methyl nutrient Bacillus inoculum in a weight ratio of (60-80):(5-15):(5-15):(3-8):(3-8). The active ingredient of the methyltrophic Bacillus agent is methyltrophic Bacillus GBacillus-9 with the preservation number CGMCC No.13337; As chickens progress from chicks to young chickens and then to adult chickens, the amount of the compound functional additive added to the feed increases at each stage. (2) Gradual grazing management based on altitude gradients: The feeding and management of chickens are divided into two stages: indoor brooding and outdoor free-range. During the indoor brooding stage, the brooding temperature is controlled according to the altitude of the breeding area, with the brooding temperature in high-altitude areas being higher than that in low-altitude areas. Before entering the outdoor free-range stage, a transition period is set up, and the outdoor activity time is gradually increased each day to allow the chickens to gradually adapt to the woodland environment. After entering the outdoor grazing stage, grazing is carried out in a zoned rotational grazing manner, with each rotational grazing area grazing for a predetermined number of days before resting.

2. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, The feeds for each stage all contain a basal diet and compound functional additives; wherein, the compound functional additives in the chick stage feed are added at 0.3%-0.7% of the basal diet weight, the compound functional additives in the pullet stage feed are added at 0.5%-1.0% of the basal diet weight, and the compound functional additives in the adult chicken stage feed are added at 0.5%-1.5% of the basal diet weight. And / or, the weight ratio of honeysuckle extract, artemisia extract, houttuynia cordata extract, allicin and methyl nutrient Bacillus agent in the compound functional additive is 70:10:10:5:

5.

3. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, The chick stage is from 0 to 40 days old, the young chicken stage is from 41 to 120 days old, and the adult chicken stage is from 121 days old until slaughter. And / or, the indoor brooding stage is from 0 to 40 days old, the transition period is from 41 to 60 days old, and the outdoor free-range stage is from 61 days old to market age.

4. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 3, characterized in that, The basic daily ration of the feed for this stage comprises, by weight percentage: 60%-68% corn, 20%-26% soybean meal, 3%-7% wheat bran, 1%-3% rice bran, 0.8%-1.2% dicalcium phosphate, 1.5%-2.5% limestone powder, and 2.5%-3.5% micronutrient premix, and the sum of the weight percentages of the above components is 100%; wherein, the micronutrient premix contains vitamin A, vitamin D, vitamin E, copper sulfate, and methionine.

5. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 4, characterized in that, In the chick stage feed, the rice bran is replaced with fish meal; And / or, starting from 31 days of age, add compound forage powder to the stage feed, said compound forage powder being a mixture of alfalfa powder and leafy grass powder in a mass ratio of (5-7):(3-5); in the chick stage feed aged 31-40 days, the amount of compound forage powder added is 0.6%-1.0% of the weight of the basic daily ration; in the pullet stage feed and the adult chicken stage feed, the amount of compound forage powder added is 1.2%-1.8% of the weight of the basic daily ration.

6. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 5, characterized in that, The preparation process of the feed for this stage includes: The basic diet is mixed according to the formula and preheated to 55℃-65℃. Then, the compound functional additives are added according to the formula and mixed evenly. The pellets are then granulated using a low-temperature steam modulation pelleting process at 65℃-75℃. The resulting pellets have a diameter of 2.5mm-4.5mm and a hardness of 2.0kgf-3.0kgf. For stage feeds without added compound forage powder, the pellets obtained from pelleting are the finished stage feed products; for stage feeds requiring the addition of compound forage powder, the compound forage powder is mixed in after the pellets have cooled, and the amount added is determined as follows: In the feed for chicks aged 31-40 days, the amount of compound forage powder added is 0.6%-1.0% of the basal diet weight; in the feed for pullets and adult chickens, the amount of compound forage powder added is 1.2%-1.8% of the basal diet weight. When adding compound forage powder, a step-by-step premixing method is adopted. First, the compound forage powder is mixed evenly with a small amount of cooled feed pellets, and then the mixture is expanded to all feed pellets in the batch until it is evenly mixed. The preparation method of the compound forage powder includes: selecting alfalfa in the early flowering stage and leafy grass in the vigorous growth stage as raw materials, harvesting them on a sunny morning, retaining the tender stems and leaves, and removing the aged lignified straw and impurities; spreading them out to air dry naturally for 4-6 hours to remove surface free moisture, and then transferring them to a cool and ventilated place to air dry until the moisture content is ≤12%; mixing the dried alfalfa and leafy grass in the specified proportion, and pulverizing them to 40-60 mesh using a low-temperature and low-speed pulverizer; after pulverizing, sieving to remove impurities, and storing them in a dry, light-proof, and sealed environment to prevent moisture absorption and mold growth.

7. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, During the indoor brooding stage: When the altitude of the breeding area is ≤1200 meters, the brooding temperature should be gradually reduced from 33℃-37℃ to 23℃-27℃, with a daily temperature decrease of 0.2℃-0.3℃, and the humidity should be controlled at 55%-65%. When the altitude of the breeding area is greater than 1200 meters, the brooding temperature should be gradually reduced from 35℃-39℃ to 25℃-29℃, with a daily temperature reduction of 0.2℃-0.3℃, and the humidity should be controlled at 55-65%.

8. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, During the transition period: when the altitude of the breeding area is below 1200 meters, the initial outdoor activity time is 2-4 hours, which is extended by 0.5-1.5 hours every day until the animals are free-range all day; when the altitude of the breeding area is above 1200 meters, the activity area is only opened when the daytime temperature is ≥12℃, and the initial outdoor activity time is 1.5-2.5 hours, which is extended by 1 hour every 2-4 days until the animals are free-range all day. During the outdoor free-range stage: when the altitude of the breeding area is below 1200 meters, multiple rotational grazing areas are divided according to the standard of 6-10 mu / 100 chickens. Each rotational grazing area is free-range for 15-25 days and rested for 35-45 days. Every 80-120 chickens are equipped with a 4㎡-6㎡ mobile chicken house. When the altitude of the breeding area is above 1200 meters, multiple rotational grazing areas are divided according to the standard of 5-8 mu / 100 chickens. Each rotational grazing area is free-range for 25-35 days and rested for 45-60 days. Every 80-120 chickens are equipped with a 4㎡-6㎡ mobile chicken house.

9. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, It also includes prevention and management measures selected from at least one of the following: (a) Immunization: 0.15-0.25 ml / animal subcutaneously for Marek's disease vaccine at 1 day old; 0.04-0.06 ml / animal intranasally for Newcastle disease vaccine at 10 days old; 0.08-0.12 g / L of infectious bursal disease vaccine administered via drinking water at 14 days old; and 0.25-0.35 ml / animal intramuscularly for Escherichia coli vaccine at 50 days old when the altitude of the breeding area is >1200 meters. (b) Biosecurity: During the free-range stage: every 20-40 days, for 2-4 consecutive days each time, administer honeysuckle decoction in drinking water at a concentration of 0.3-0.7 g / L, with the water temperature controlled at 22-28℃; disinfect the chickens with 0.3% povidone-iodine every 15 days; deworm with thiabendazole once a month; when changing to a rotational grazing area, place a 3-7 cm thick layer of quicklime and disinfect the chickens by stepping on the basin. (c) Control of natural enemies: Hang reflective strips or old CDs every 1.5-2.5 meters; erect a nylon mesh fence with a height of 2.0-2.4 meters and a mesh size of (4-6) cm × (4-6) cm around the breeding area; prepare a powder by mixing realgar and wood ash in a weight ratio of 1:(2-4), and spread a protective strip 0.3-0.7 meters wide along the inside of the fence once a month, and respread it after rain; (d) Human interference control: A wooden fence with a width of 40-60 meters and a height of 1.5-2.0 meters shall be set up between the breeding area and the external road; feeding shall be carried out at fixed times from 6:30 to 7:30 a.m. and from 5:30 p.m. to 6:30 p.m. every day, and feeding equipment shall be cleaned and disinfected once a week; (e) Monitoring measures: High-definition surveillance cameras will be installed in the foraging areas and water sources of the rotational grazing area, and real-time monitoring will be conducted via a mobile app; (f) Portable chicken coop structure: equipped with slatted flooring, including: Chicken houses in low-altitude areas: shoulder height 1.8-2.0m, ridge height 3.2-3.5m, walls are covered with double layers of insulation cotton and waterproof membrane, and the roof is equipped with openable ventilation windows; Chicken houses in high-altitude areas: shoulder height 2.0-2.2m, ridge height 3.5-3.8m, wall insulation layer thickness is greater than that of chicken houses in low-altitude areas, and double-layer tempered glass is used for lighting; All slatted floors have gaps 1.5cm-2.0cm wide and are made of anti-corrosion pine wood. A removable manure collection trough is installed underneath, with the trough tilted at an angle of 5°-8°. Inside the chicken house, there is one feeding trough and one waterer for every 8-12 chickens, and multi-level perches are provided 40cm-50cm off the ground. The location of the chicken house should avoid low-lying, waterlogged areas in mountainous areas. Low-altitude chicken houses should have additional drainage ditches, while high-altitude chicken houses should have their bottom raised by 25cm-35cm.

10. The method for ecological breeding of mountain plum blossom chickens adapted to altitude gradients according to claim 1, characterized in that, When chickens are raised to 140-160 days old and ready for market, their weight should be ≥1.49kg, feed conversion ratio ≤19.43%, breast muscle shear force ≤26.26N, leg muscle drip loss rate ≤2.09%, and breast muscle drip loss rate ≤1.93%.

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