Compound feed additive for improving semen quality of cocks in high-altitude area and preparation method of compound feed additive

Abstract

The invention provides a compound feed additive for improving the semen quality of cocks in high-altitude areas and a preparation method of the compound feed additive. Polyvinyl alcohol-trehalose-succinate is used as a nanofiber skeleton, and the surface of the nanofiber skeleton is modified with a hyaluronic acid-polydopamine coating. Loading antioxidant components (chlorogenic acid and vitamin E), anti-hypoxia components (cordycepin and salidroside) and spermatogenic nutritional components (L-arginine and hermetia illucens insect powder) step by step, and finally mixing with carriers (zeolite powder and defatted rice bran). The additive can respond to stress signals such as free radical concentration increase, hypoxia and temperature difference in the plateau environment, target release of functional components is achieved, the plateau specific oxidative stress-tissue hypoxia-energy metabolism disorder chain reaction is relieved synergistically from the source, the spermatogenic function of testis is accurately improved, the sperm density and vitality of cocks are effectively improved, and the production efficiency of the cocks is improved. Finally, the fertilization rate and the hatching rate of hatching eggs are improved.

Description

Technical Field

[0001] This invention belongs to the field of animal feed technology, specifically relating to a compound feed additive for improving the quality of rooster semen in high-altitude areas and its preparation method. Background Technology

[0002] The unique environmental stresses of high-altitude regions (typically above 2000 meters), including persistent low oxygen levels, strong ultraviolet radiation, and large diurnal temperature variations, pose a severe challenge to livestock and poultry farming, particularly impacting the reproductive performance of breeding animals. For breeding roosters, the high-altitude, low-oxygen environment triggers a sharp increase in oxidative stress, leading to decreased blood circulation efficiency and insufficient oxygen supply to the testicular tissue. This series of physiological disturbances severely disrupts spermatogenesis and endocrine balance, manifesting as a significant decrease in sperm density, sperm motility, sperm abnormality rate, and semen volume. This deterioration in key semen parameters directly results in a sharp decline in the fertilization and hatching rates of hatching eggs, causing sustained economic losses to poultry farming in high-altitude areas.

[0003] To improve the reproductive performance of breeding roosters, current technologies commonly employ the addition of conventional nutrients or antioxidants to their feed, such as supplementing with vitamins (e.g., VE, VC), trace elements (e.g., selenium, zinc), and amino acids (e.g., arginine). However, these conventional methods, proven effective in lowland areas, often prove inadequate and ineffective in addressing the multi-stage, cascading complex stress challenges unique to high-altitude regions, namely "oxidative stress-tissue hypoxia-energy metabolism disorders." The fundamental reason is that conventional additives are mostly single or simple combinations of functional components with dispersed targets, failing to systematically respond to and block the aforementioned chain of pathophysiological responses induced by the high-altitude environment, and also unable to achieve precise nutritional regulation of the reproductive system.

[0004] Although some natural active substances have shown potential in addressing specific stresses—for example, chlorogenic acid possesses excellent antioxidant and anti-inflammatory capabilities, cordycepin derived from high-altitude medicinal fungi has been proven to enhance tissue hypoxia tolerance, and novel protein source black soldier fly powder is rich in antimicrobial peptides and essential fatty acids that support immunity and energy supply—current technologies have not yet been able to scientifically integrate and synergistically design these functional components targeting different stressors at high altitudes. More critically, there is a lack of an effective delivery system capable of intelligently controlling the targeted release of these functional components into the reproductive system based on high-altitude stress signals (such as increased free radical concentration, pH changes, and temperature differences). Therefore, developing a specialized feed additive that can systematically address the complex stresses of high altitudes and precisely act on the reproductive system of breeding roosters to improve their semen quality has become a pressing technical challenge in this field. Summary of the Invention

[0005] Technical Problem to be Solved: To address the aforementioned technical problems, the purpose of this invention is to provide a compound feed additive for improving the quality of rooster semen in high-altitude areas and its preparation method. This compound feed additive uses polyvinyl alcohol-trehalose-succinate as a nanofiber framework, with a surface modified by a hyaluronic acid-polydopamine coating. It is then loaded stepwise with antioxidant components (chlorogenic acid, vitamin E), hypoxia-resistance components (cordycepin, rhodioloside), and spermatogenic nutrients (L-arginine, black soldier fly larvae powder), and finally mixed with a carrier (zeolite powder, defatted rice bran). This additive can respond to stress signals such as increased free radical concentration, low oxygen, and temperature differences in high-altitude environments, achieving intelligent and targeted release of functional components. It synergistically alleviates the unique high-altitude chain reaction of "oxidative stress-tissue hypoxia-energy metabolism disorder," precisely improving testicular spermatogenesis, effectively increasing rooster sperm density and motility, reducing the deformity rate, and ultimately improving the fertilization and hatching rates of hatching eggs.

[0006] Technical solution: A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas, comprising the following steps: S1. Dissolve polyvinyl alcohol in deionized water and stir at 80-90℃ to obtain a polyvinyl alcohol solution with a concentration of 8-12%. After cooling to 60-70℃, add trehalose and stir to dissolve. Then add succinic anhydride and adjust the pH to 7.5. React for 2-4 hours, electrospin, vacuum dry at 40℃ for 6 hours, and pulverize through a 100-mesh sieve to obtain a nanofiber skeleton. S2. Dissolve hyaluronic acid in pH 8.5 Tris-HCl buffer solution and stir to obtain a hyaluronic acid solution with a concentration of 0.3-0.5%. Then add dopamine hydrochloride and stir at room temperature in the dark for 20-24 hours to obtain a composite solution. Immerse the nanofiber skeleton in the composite solution and shake at 30-35℃ for 10-12 hours. Remove, wash, and vacuum dry at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and the modified nanofiber framework was added. The mixture was shaken at 30-35°C in the dark for 4-6 hours, filtered, the precipitate was collected, and the mixture was vacuum dried at 40°C for 3 hours to obtain the modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water, mix with the modified nanofiber framework loaded with antioxidant components, shake for 8-10 hours at pH 6.5 and 40-50℃, filter, collect the precipitate, freeze dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant and hypoxia-resistant components. After reacting at room temperature for 2-3 hours, the mixture was filtered, the precipitate was collected, and vacuum dried at 45°C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber complex, stir and mix for 20-30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0007] Furthermore, in step S1, the mass ratio of trehalose, succinic anhydride, and polyvinyl alcohol is (0.2-0.6):(0.2-0.5):1; the electrospinning parameters are: voltage 15-18kV, receiving distance 10-20cm, flow rate 0.5-1.5mL / h, temperature 20-30℃, and humidity 30-40%.

[0008] Furthermore, in step S2, the mass ratio of dopamine hydrochloride to hyaluronic acid is (0.3-0.8):1; and the mass-volume ratio of the nanofiber skeleton to the composite solution is 1g:(15-30)mL.

[0009] Furthermore, in step S3, the mass-to-volume ratio of chlorogenic acid, vitamin E, modified nanofiber skeleton and anhydrous ethanol is (0.03-0.08) g: (0.02-0.05) g: 1 g: (4-6) mL.

[0010] Furthermore, in step S4, the mass-to-volume ratio of cordycepin, rhodioloside, modified nanofiber skeleton loaded with antioxidant components, and deionized water is (0.02-0.06) g: (0.04-0.1) g: 1 g: (2.5-4) mL.

[0011] Furthermore, in step S5, the mass-to-volume ratio of L-arginine, the modified nanofiber framework loaded with antioxidant-oxygen-resistant components, and deionized water is (0.08-0.15) g:1 g:(2-2.5) mL.

[0012] Furthermore, in step S6, the mass ratio of black soldier fly larvae ultrafine powder, zeolite powder, defatted rice bran, and functionalized nanofiber composite is (0.15-0.3):(0.4-0.6):(0.2-0.4):1.

[0013] The compound feed additive prepared by the above preparation method.

[0014] The above-mentioned compound feed additives are used in the preparation of complete feed to improve the semen quality and hatchability of roosters in high-altitude areas.

[0015] Furthermore, the compound feed additive is added to the complete feed at a rate of 0.5-3%.

[0016] Beneficial effects: This invention prepares a compound feed additive that is beneficial for improving the semen quality of roosters in high-altitude areas. This additive focuses on the modification of the nanofiber network matrix and the targeted binding of functional components. All functional components are anchored to the nanofibers through specific interactions, releasing and taking effect only upon triggering high-altitude stress signals. Specifically, a modified coating is formed by cross-linking hyaluronic acid and polydopamine to the fiber surface using polyvinyl alcohol-trehalose-succinate as the fiber backbone. Next, antioxidant components (chlorogenic acid, vitamin E) are anchored to the surface sites of the polydopamine coating through hydrophobic interactions. Hypoxia-resistance components (cordycepin, rhodioloside) are embedded into the internal sites of the trehalose-succinate backbone through hydrogen bonds. Finally, spermatogenic nutrients (L-arginine, black soldier fly larvae ultrafine powder) are blended into the fiber backbone. The compound feed additive prepared in this invention is targeted and responsive when feeding roosters in high-altitude areas. Regarding targeting, the hydrophilic groups of hyaluronic acid can enhance the compatibility between fibers and the intestinal mucus layer, allowing the nanofiber network to colonize and remain in the intestinal absorption area (ileum) for a longer period of time. This avoids the problems of decreased digestive enzyme activity and insufficient absorption caused by the low temperature at high altitudes, allowing sufficient time for antioxidant, hypoxia-resistant, and nutrient components to be absorbed into the bloodstream. Hyaluronic acid receptors mediate the targeted delivery of L-arginine to testicular tissue, improving the utilization rate of spermatogenic raw materials. The medium-chain fatty acids of black soldier flies provide rapid energy, and the antimicrobial peptides enhance immunity, adapting to the needs of high-altitude stress. The compound feed additive prepared in this invention exhibits responsiveness to roosters fed at high altitudes, specifically including free radical concentration response and low oxygen / temperature difference response. ① Free radical concentration response: Strong ultraviolet radiation at high altitudes leads to a free radical concentration ≥150 U / mL in the roosters. When the free radical concentration in the roosters increases, polydopamine preferentially undergoes redox reactions with free radicals, causing the benzene ring in its molecular structure to open, the coating sites to swell, and chlorogenic acid and vitamin E anchored on the surface to be rapidly released, avoiding ineffective release under no-stress conditions. Precisely protects sperm cell membranes; ② Response to hypoxia and temperature difference: In the hypoxic environment of the plateau, the lactic acid content in the rooster's body increases, leading to a decrease in intestinal pH. The ester bonds of succinate in the fibrous skeleton break under acidic conditions, and the skeleton structure begins to swell. When the diurnal temperature difference on the plateau is ≥15℃, the crystal structure of trehalose changes from anhydrous to dihydrate, expands in volume, further damages the skeleton structure, accelerates the release of cordycepin and rhodioloside. Rhodioloside enhances the oxygen-carrying capacity of red blood cells, and cordycepin improves mitochondrial energy supply, targeting and solving the problem of testicular hypoxia. The compound feed additive prepared by this invention has all components that act precisely on the reproductive system through specific response and targeted delivery, thereby restoring testicular spermatogenesis from the root, increasing sperm density and motility, reducing the deformity rate, and thus improving the fertilization rate and hatching rate of hatching eggs. Detailed Implementation

[0017] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments: Example 1

[0018] A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.3:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:15mL and shaken at 35℃ for 12 hours. The solution was then removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.03 g: 0.02 g: 1 g: 4 mL. The mixture was shaken at 30 °C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40 °C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.02 g:0.04 g:1 g:2.5 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.08 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.15:0.4:0.2:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0019] Example 2 A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.8:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:30mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.03 g: 0.02 g: 1 g: 4 mL. The mixture was shaken at 30 °C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40 °C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.02 g:0.04 g:1 g:2.5 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.08 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.15:0.4:0.2:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0020] Example 3 A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.03 g: 0.02 g: 1 g: 4 mL. The mixture was shaken at 30 °C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40 °C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.02 g:0.04 g:1 g:2.5 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.08 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.15:0.4:0.2:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0021] Example 4 A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.06 g:0.03 g:1 g:6 mL. The mixture was shaken at 30°C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40°C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.04 g:0.07 g:1 g:3 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.08 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.15:0.4:0.2:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0022] Example 5 A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.06 g:0.04 g:1 g:6 mL. The mixture was shaken at 30°C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40°C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.04 g:0.08 g:1 g:4 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.1 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran. After mixing the zeolite powder and defatted rice bran, add them to the functionalized nanofiber composite in a mass ratio of 0.2:0.4:0.2:1. Stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain the compound feed additive.

[0023] Example 6 A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.06 g:0.04 g:1 g:6 mL. The mixture was shaken at 30°C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40°C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.04 g:0.08 g:1 g:4 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.1 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.2:0.4:0.4:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0024] Comparative Example 1 The difference between this comparative example and Example 6 is that the nanofiber framework was not modified, as detailed below: A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol and a nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, nanofiber framework and anhydrous ethanol was 0.06 g:0.04 g:1 g:6 mL. The mixture was shaken at 30 °C in the dark for 4 h, filtered, the precipitate was collected and dried under vacuum at 40 °C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S3. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.04 g:0.08 g:1 g:4 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze-dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S4. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant-antioxidant-antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with antioxidant-antioxidant components and deionized water was 0.1 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S5. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.2:0.4:0.4:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0025] Comparative Example 2 The difference between this comparative example and Example 6 is that it does not contain an antioxidant component, as detailed below: A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber skeleton, wherein the mass-volume ratio of cordycepin, rhodioloside, modified nanofiber skeleton and deionized water is 0.04g:0.08g:1g:4mL. Shake for 10h at pH 6.5 and 45℃, filter, collect the precipitate and freeze dry to obtain the modified nanofiber skeleton loaded with hypoxia-resistant components. S4. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with hypoxia-resistant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber framework loaded with hypoxia-resistant components and deionized water was 0.1 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S5. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.2:0.4:0.4:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0026] Comparative Example 3 The difference between this comparative example and Example 6 is that it does not contain an oxygen-tolerant component, as detailed below: A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.06 g:0.04 g:1 g:6 mL. The mixture was shaken at 30°C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40°C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. L-arginine was dissolved in deionized water and mixed with the modified nanofiber skeleton loaded with antioxidant components. After reacting at room temperature for 3 hours, the mass-volume ratio of L-arginine, modified nanofiber skeleton loaded with antioxidant components and deionized water was 0.1 g:1 g:2 mL. The mixture was stirred evenly, filtered, the precipitate was collected, and vacuum dried at 45 °C for 3 hours to obtain the functionalized nanofiber complex. S5. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.2:0.4:0.4:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0027] Comparative Example 4 The difference between this comparative example and Example 6 is that it does not contain spermatogenic nutrients, as detailed below: A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas includes the following steps: S1. Polyvinyl alcohol was dissolved in deionized water and stirred at 90°C to obtain a 10% polyvinyl alcohol solution. After cooling to 60°C, trehalose was added and stirred to dissolve. Then, succinic anhydride was added. The mass ratio of trehalose, succinic anhydride and polyvinyl alcohol was 0.4:0.3:1. The pH was adjusted to 7.5, and the reaction was carried out for 3 hours. Electrospinning was performed (voltage 15kV, receiving distance 20cm, flow rate 1.0mL / h, temperature 25°C, humidity 40%). The solution was then vacuum dried at 40°C for 6 hours and pulverized through a 100-mesh sieve to obtain a nanofiber framework. S2. Hyaluronic acid was dissolved in pH 8.5 Tris-HCl buffer solution and stirred to obtain a 0.5% hyaluronic acid solution. Dopamine hydrochloride (mass ratio of 0.6:1 to hyaluronic acid) was then added and stirred at room temperature in the dark for 24 hours to obtain a composite solution. The nanofiber skeleton was immersed in the composite solution at a mass-volume ratio of 1g:20mL, shaken at 35℃ for 12 hours, removed, washed, and vacuum dried at 45℃ for 4 hours to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and a modified nanofiber framework was added. The mass-volume ratio of chlorogenic acid, vitamin E, modified nanofiber framework and anhydrous ethanol was 0.06 g:0.04 g:1 g:6 mL. The mixture was shaken at 30°C in the dark for 4 h, filtered, the precipitate was collected, and vacuum dried at 40°C for 3 h to obtain a modified nanofiber framework loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water and mix with the modified nanofiber framework loaded with antioxidant components. The mass-volume ratio of cordycepin, rhodioloside, modified nanofiber framework loaded with antioxidant components and deionized water is 0.04 g:0.08 g:1 g:4 mL. Shake for 10 h at pH 6.5 and 45 °C. Filter, collect the precipitate, and freeze dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components, i.e., the functionalized nanofiber complex. S5. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber composite, with a mass ratio of 0.2:0.4:0.4:1, stir and mix for 30 minutes, and pass through an 80-mesh sieve to obtain compound feed additive.

[0028] Performance testing: 1. Selection and grouping of experimental animals Two hundred and twenty healthy Hy-Line Brown parent roosters aged 44 weeks were selected from a chicken farm in a high-altitude area. After being balanced according to body weight, semen volume, sperm motility and sperm density, they were divided into a control group and an experimental group, for a total of 11 groups, with four replicates in each group and five roosters in each replicate.

[0029] 2. Diet and Feeding Management A basal diet was prepared according to the nutritional requirements of breeding roosters as specified by the NRC and Chinese chicken feeding standards. The control group was fed the basal diet, while the experimental group was fed an experimental diet supplemented with 0.5% of the compound feed additives for improving rooster semen quality (Examples 1-6 and Comparative Examples 1-4) from the basal diet. During the experiment, both groups had free access to feed and water and received routine immunizations. After a one-week trial period of feeding both the control and experimental groups with the basal diet, the experimental group began to be fed the experimental diet, while the control group continued to be fed the basal diet. The experimental period was 5 weeks, from 45 to 50 weeks of age.

[0030] 3. Data Collection and Index Measurement ① Measurement of ejaculation volume: The collection cup is preheated. After collecting semen by abdominal massage, the volume of semen is directly read on the collection cup. ②Density determination: The semen diluted with 0.3% NaCl solution was dropped into the hemocytometer chamber using a dropper (the semen around the coverslip was wiped off with lens paper, and the coverslip was made free of air bubbles). The semen was then observed under a high-power microscope. The number of sperm in the five squares (four corners and the center) was recorded. Only the number of sperm in more than 3 / 4 of the squares was counted. Sperm in larger than 3 / 4 of the squares were counted. Methods for calculating semen density: Sperm count per milliliter = Total sperm count in 5 squares × 5 × 10 × 1000 × Dilution factor ③ Sperm motility determination: Use a dropper to draw up semen diluted with 0.3% NaCl solution, drop it onto a glass slide, cover it with a coverslip, ensuring there are no air bubbles, observe it under a microscope, and count the number of sperm that can move in a straight line within one field of view; Methods for calculating sperm motility: Sperm motility = (Number of sperm moving in a straight line / Total number of sperm in the field of view) × 100% ④ Determination of sperm abnormality rate: Semen smears were taken, stained with Giemsa stain, and abnormal sperm were examined under a microscope. Two smears were made from each sample, and 500 sperm were counted on each smear. The abnormality rate was calculated. Criteria for determining abnormal sperm: Abnormal sperm mainly refer to giant heads, small heads, defects, double heads, pear-shaped heads, protruding heads, enlarged necks, enlarged mitochondria, presence of protoplasmic particles, taillessness, curled tails, and other curled features; Methods for calculating sperm abnormality rate: Sperm abnormality rate (%) = Number of abnormal sperm / 500 × 100; Table 1

[0031] As shown in Table 1, adding the compound feed additive provided by this invention to the diet of breeding roosters in high-altitude environments can effectively improve various semen quality indicators. However, Comparative Example 1 lacks a hyaluronic acid-polydopamine coating, and the functional components are only simply loaded, which cannot achieve precise release under stress triggering and targeted enrichment of the reproductive system, resulting in low utilization efficiency and a significant reduction in overall effect. Comparative Example 2 lacks antioxidant components, and its sperm abnormality rate is significantly higher than that of Example 6, indicating that in a strong ultraviolet environment, the lack of targeted antioxidant protection makes sperm cell membranes and DNA more susceptible to damage. Comparative Example 3 lacks hypoxia-resistant components, and its sperm density and motility improvement are limited, indicating that under continuous low oxygen conditions, antioxidants alone cannot improve the energy metabolism and oxygen supply of testicular parenchymal cells, thus hindering the improvement of spermatogenesis efficiency. Comparative Example 4 lacks spermatogenesis nutrients, and its semen volume, density, and other indicators are better than other comparative examples, but its sperm motility and abnormality rate are still relatively high. This indicates that in a high-altitude stress environment, the spermatogenesis process requires additional raw materials, such as L-arginine and black soldier fly larvae powder, and simple stress protection is insufficient to maximize reproductive performance.

[0032] 4. Determination of fertilization rate and hatchability of fertilized eggs 1) Fertilization rate of hatching eggs: ① Egg Collection: Healthy Hy-Line Brown parent hens were raised in the same groups and replicates as the roosters (i.e., divided into control and experimental groups, a total of 11 groups, 4 replicates per group, with 5 hens per replicate). Roosters in each replicate were kept separately from their corresponding hens or bred using artificial insemination to ensure that the hatching eggs corresponded to the specific experimental group. During the experimental period (45 to 50 weeks of age), hatching eggs from the corresponding hens in each group were collected daily and labeled according to group and replicate (labeling group, replicate number, and laying date). Broken, cracked, deformed, and excessively large / small eggs (eggs that did not meet the hatching egg standards) were removed to ensure a consistent number of hatching eggs per replicate in each group (at least 30 hatching eggs were collected per replicate in each group, with the total number of hatching eggs meeting the requirements of the hatching experiment). ② Egg pretreatment: Place the collected eggs in an environment of 20-25℃ and 60-70% relative humidity for 12-24 hours to allow the internal temperature of the eggs to balance with the ambient temperature, and avoid excessive temperature difference in the early stage of incubation affecting embryo development; after pretreatment, wipe the surface of the eggs with 0.1% potassium permanganate solution for disinfection, and then let them dry for later use. ③ Incubation and candling: Place the pretreated eggs from each group into the same incubator and incubate them uniformly according to the standard incubation conditions for Hy-Line Brown eggs (temperature 37.8℃, relative humidity 55-60% for the first 7 days of incubation; temperature 37.5℃, relative humidity 50-55% for days 8-18 of incubation; temperature 37.2℃, relative humidity 65-70% for days 19-21 of incubation). Turn the eggs 6 times a day (once every 4 hours) to ensure consistent incubation conditions. ④ Fertilization determination and counting: On the 7th day of incubation, the eggs are candled for the first time using an egg candling device to observe the embryonic development inside the eggs and determine whether they are fertilized or unfertilized. Criteria for determining fertilized eggs: During candling, a clear embryonic shadow, a clear vascular network, and a heartbeat can be seen inside the egg; Criteria for identifying unfertilized eggs: During candling, the egg is transparent with no embryonic shadow, only the yolk is visible floating, and there is no blood vessel development; ⑤ Calculation method: Count the number of fertilized eggs and the number of hatching eggs in each group and each replicate, and calculate the fertilization rate of the hatching eggs: Fertilization rate of hatching eggs (%) = (Total number of fertilized eggs per group / Total number of hatching eggs incubated per group) × 100% 2) Hatching rate of fertilized eggs: ①Continued incubation: After candling, remove unfertilized eggs and dead embryos, and continue incubating the remaining fertilized eggs under the above-mentioned standard incubation conditions until hatching on the 21st day of incubation; ② Hatching count: During the hatching period (days 19-21 of incubation), observe the hatching situation daily and promptly remove healthy chicks, weak chicks (listless, underweight, or with limb deformities) and dead embryos (embryos that did not hatch or died after hatching). Count the number of healthy chicks, weak chicks, and dead embryos for each group and each replicate. ③ Hatching rate determination: The hatching rate of fertilized eggs is based on the number of healthy chicks, and weak chicks and dead embryos are excluded to ensure that the data reflects the true hatching effect; Criteria for judging healthy chicks: lively and energetic, with clean and glossy feathers, well-healed navel without bleeding, intact limbs, and able to stand and eat normally; weak chicks and dead embryos are not counted as healthy chicks; ④ Calculation method: Count the number of healthy chicks and the total number of fertilized eggs in each group, and calculate the hatching rate of the fertilized eggs: Hatching rate of fertilized eggs (%) = (Total number of healthy chicks in each group / Total number of fertilized eggs in each group) × 100% Table 2

[0033] As shown in Table 2, the compound feed additive prepared in the embodiments of the present invention improved the fertilization rate and hatchability of fertilized eggs after feeding breeding roosters. However, Comparative Example 1 lacked a hyaluronic acid-polydopamine coating, and the functional components could not achieve precise release and targeted enrichment of the reproductive system under stress triggering, resulting in low utilization efficiency. Comparative Example 2 lacked antioxidant components, and under high-altitude and strong ultraviolet environment, sperm cell membranes and DNA were more easily damaged, resulting in a higher sperm abnormality rate, which limited the improvement of fertilization rate and hatchability. Comparative Example 3 lacked hypoxia-resistant components, and the improvement of energy metabolism and oxygen supply in testicular parenchymal cells was limited, hindering spermatogenesis efficiency. Therefore, the improvement of sperm density and motility was insufficient, affecting fertilization potential. Comparative Example 4 lacked spermatogenesis nutrients. Although it had a certain stress protection effect, the raw materials for the spermatogenesis process were insufficient, resulting in insufficient improvement of sperm motility and abnormality rate, and relatively low fertilization rate and hatchability.

[0034] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A method for preparing a compound feed additive to improve the quality of rooster semen in high-altitude areas, characterized in that, Includes the following steps: S1. Dissolve polyvinyl alcohol in deionized water and stir at 80-90℃ to obtain a polyvinyl alcohol solution with a concentration of 8-12%. After cooling to 60-70℃, add trehalose and stir to dissolve. Then add succinic anhydride, adjust the pH to 7.5, react for 2-4 hours, electrospin, vacuum dry, pulverize and sieve to obtain a nanofiber skeleton. S2. Dissolve hyaluronic acid in pH 8.5 Tris-HCl buffer solution and stir to obtain a hyaluronic acid solution with a concentration of 0.3-0.5%. Then add dopamine hydrochloride and stir at room temperature in the dark for 20-24 hours to obtain a composite solution. Immerse the nanofiber skeleton in the composite solution and shake at 30-35℃ for 10-12 hours. Remove, wash, and vacuum dry to obtain the modified nanofiber skeleton. S3. Chlorogenic acid and vitamin E were dissolved in anhydrous ethanol, and the modified nanofiber skeleton was added. The mixture was shaken at 30-35°C in the dark for 4-6 hours. The mixture was then filtered, the precipitate was collected, and the mixture was vacuum dried to obtain the modified nanofiber skeleton loaded with antioxidant components. S4. Dissolve cordycepin and rhodioloside in deionized water, mix with the modified nanofiber framework loaded with antioxidant components, shake for 8-10 hours at pH 6.5 and 40-50℃, filter, collect the precipitate, freeze dry to obtain the modified nanofiber framework loaded with antioxidant components and hypoxia-resistant components. S5. L-arginine was dissolved in deionized water and mixed with the modified nanofiber framework loaded with antioxidant and hypoxia-resistant components. After reacting at room temperature for 2-3 hours, the mixture was filtered, the precipitate was collected, and vacuum dried to obtain the functionalized nanofiber complex. S6. Mix black soldier fly insect ultrafine powder with zeolite powder and defatted rice bran, add to functionalized nanofiber complex, stir and mix for 20-30 minutes, and sieve to obtain compound feed additive.

2. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S1, the mass ratio of trehalose, succinic anhydride, and polyvinyl alcohol is (0.2-0.6):(0.2-0.5):1; the electrospinning parameters are: voltage 15-18kV, receiving distance 10-20cm, flow rate 0.5-1.5mL / h, temperature 20-30℃, and humidity 30-40%.

3. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S2, the mass ratio of dopamine hydrochloride to hyaluronic acid is (0.3-0.8):1; the mass-volume ratio of the nanofiber skeleton to the composite solution is 1g:(15-30)mL.

4. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S3, the mass-to-volume ratio of chlorogenic acid, vitamin E, modified nanofiber skeleton and anhydrous ethanol is (0.03-0.08) g: (0.02-0.05) g: 1 g: (4-6) mL.

5. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S4, the mass-to-volume ratio of cordycepin, rhodioloside, modified nanofiber skeleton loaded with antioxidant components, and deionized water is (0.02-0.06) g: (0.04-0.1) g: 1 g: (2.5-4) mL.

6. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S5, the mass-to-volume ratio of L-arginine, modified nanofiber skeleton loaded with antioxidant-oxygen-resistant components, and deionized water is (0.08-0.15) g:1 g:(2-2.5) mL.

7. The method for preparing a compound feed additive for improving the quality of rooster semen in high-altitude areas according to claim 1, characterized in that, In step S6, the mass ratio of black soldier fly insect ultrafine powder, zeolite powder, defatted rice bran, and functionalized nanofiber composite is (0.15-0.3):(0.4-0.6):(0.2-0.4):

1.

8. The compound feed additive prepared by the preparation method according to any one of claims 1-7.

9. The application of the compound feed additive prepared according to claim 8 in the preparation of complete feed to improve the quality of rooster semen and hatchability in high-altitude areas.

10. The application according to claim 9, characterized in that: The compound feed additive is added at a rate of 0.5-3% in the complete feed.

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