Biological additive for promoting feather development of poultry and application of biological additive

Abstract

The invention discloses a biological additive for promoting poultry feather development and application thereof, and belongs to the technical field of animal breeding. The biological additive is prepared from scallop skirt oligopeptide and stiff silkworm polypeptide according to the mass ratio of 1: 1. Animal experiments show that when 60 mg of the biological additive is added into every 1 kg of basic feed to feed broilers, the growth of the broilers can be promoted, and more importantly, the feather development of the broilers can be remarkably and synergistically promoted, and the length of main wing feather is obviously increased. The cytology research further proves that the biological additive can synergistically promote the proliferation of the poultry hair follicle epidermal stem cells, and the effect of the biological additive is superior to that of EGF positive control. Therefore, the biological additive developed by the invention is safe, efficient and green, effectively solves the problems of drug residue and drug resistance possibly caused by the traditional chemical additive, provides a sustainable solution for poultry breeding, and has important economic and application values.

Description

Technical Field

[0001] The invention belongs to the technical field of animal breeding, and in particular relates to a biological additive for promoting poultry feather development and application thereof. Background Art

[0002] In poultry farming, feather growth is not only a key indicator of health but also directly impacts the economic benefits of the operation. For poultry, well-maintained feathers not only help maintain normal body temperature and support flight, but also, to a certain extent, reflect the quality of their appearance, positively impacting breeding selection and the market value of commercial birds.

[0003] Currently, common methods used within the industry to promote feather development in poultry include nutritional regulation, environmental management, and the use of some synthetic chemical additives. However, these traditional methods generally have limitations. For example, while some synthetic chemical additives can improve feather quality in the short term, long-term use can pose a series of problems. Firstly, they pose the risk of drug residues, impacting the food safety of poultry meat and eggs. Secondly, they can lead to an imbalance in the poultry's microbiome and even cause drug resistance, thus impacting the overall sustainable development of the breeding ecosystem.

[0004] Therefore, there is an urgent need to develop a new type of feather development-promoting preparation that is safe, efficient, green and has comprehensive regulatory functions, so as to effectively promote the healthy growth of feathers while taking into account nutritional supply and physiological regulation, and avoid adverse effects on the environment and human health. Summary of the Invention

[0005] The purpose of the present invention is to provide a biological additive for promoting the development of poultry feathers and an application thereof, thereby safely and effectively promoting the growth of poultry feathers and increasing their economic value.

[0006] To achieve the above object, the present invention provides the following technical solutions: In a first aspect, the present invention provides a biological additive for promoting poultry growth and feather development, wherein the biological additive is composed of scallop skirt oligopeptide and Bombyx batryticatus polypeptide, and the mass ratio of the scallop skirt oligopeptide to the Bombyx batryticatus polypeptide is 1:1.

[0007] Preferably, the preparation method of the scallop skirt oligopeptide comprises the following steps: (1) After cleaning the scallop skirt, blanch it in boiling water, cool it in ice water and drain it; (2) freeze-drying and then crushing and sieving to obtain scallop skirt powder; (3) Deionized water was added to the scallop skirt powder at a solid-liquid ratio of 1:10, and the mixture was extracted at 4°C for 12 hours to obtain a scallop skirt aqueous extract; (4) adjusting the pH of the scallop skirt water extract to 7.5 and 55° C., adding a composite protease for enzymatic hydrolysis for 5 hours, wherein the composite protease comprises flavor protease, alkaline protease, and trypsin in a mass ratio of 5:3:2; (5) After inactivation and centrifugation, the enzymatic hydrolysate was filtered using 100 kDa, 10 kDa, and 1 kDa ultrafiltration membranes, and the 1 kDa permeate was collected; (6) The 1 kDa permeate was freeze-dried to obtain scallop skirt oligopeptide.

[0008] Preferably, the method for preparing the Bombyx batryticatus polypeptide comprises the following steps: (a) Bombyx batryticatus was pre-frozen in liquid nitrogen, ground into powder, and defatted with petroleum ether. (b) Deionized water was added at a solid-to-liquid ratio of 1:10 and extracted overnight at 4°C to obtain a water extract; (c) adjusting the aqueous extract to pH 7.5 and 55° C., and adding a composite protease for enzymatic hydrolysis for 5 hours, wherein the composite protease comprises flavor protease, alkaline protease, and trypsin in a mass ratio of 5:3:2; (d) The enzymatic hydrolysate was inactivated and centrifuged, and then purified by Sephadex G-50 gel chromatography column. The fractions corresponding to the absorption peak at 280 nm were collected and lyophilized to obtain Bombyx batryticatus polypeptide.

[0009] Preferably, the amount of the biological additive used for feed addition is 60 mg per 1 kg of basic feed; The biological additive achieves the effect of promoting poultry feather development by promoting the proliferation of poultry hair follicle epidermal stem cells.

[0010] Preferably, the poultry is chicken.

[0011] Preferably, the type of chicken is AA broiler chicken.

[0012] In a second aspect, the present invention provides a use of a biological additive composed of a scallop skirt oligopeptide and a silkworm polypeptide in preparing a feed for synergistically promoting poultry feather development, characterized in that the scallop skirt oligopeptide is prepared by the above-mentioned preparation method of the scallop skirt oligopeptide; The Bombyx batryticatus polypeptide is prepared by the above-mentioned preparation method of Bombyx batryticatus polypeptide; In the biological additive, the mass ratio of the scallop skirt oligopeptide to the silkworm polypeptide is 1:1.

[0013] Preferably, the feed consists of basic feed and biological additives; In the feed, the usage amount of the biological additive corresponding to each 1 kg of the basic feed is 60 mg.

[0014] Preferably, the poultry is chicken.

[0015] Preferably, the type of chicken is AA broiler chicken.

[0016] In a third aspect, the present invention provides a feed for promoting poultry growth and feather development, the feed comprising a basic feed and the biological additive; In the feed, the usage amount of the biological additive corresponding to each 1 kg of the basic feed is 60 mg.

[0017] Preferably, the poultry is chicken.

[0018] Preferably, the type of chicken is AA broiler chicken.

[0019] In a fourth aspect, the present invention provides a use of a biological additive composed of a scallop skirt oligopeptide and a silkworm polypeptide in preparing a cell culture preparation for synergistically promoting the proliferation of poultry hair follicle epidermal stem cells, wherein the scallop skirt oligopeptide is prepared by the above-mentioned preparation method of the scallop skirt oligopeptide; The Bombyx batryticatus polypeptide is prepared by the above-mentioned Bombyx batryticatus polypeptide preparation method; In the biological additive, the mass ratio of the scallop skirt oligopeptide to the silkworm polypeptide is 1:1.

[0020] Preferably, the biological preparation consists of biological additives, DMEM basal medium and 10% fetal bovine serum; In the biological preparation, the added concentration of the biological additive is 20 μg / mL.

[0021] Preferably, the poultry is chicken.

[0022] Preferably, the type of chicken is AA broiler chicken.

[0023] The beneficial effects of the present invention are: The present invention provides a biological additive that synergistically promotes poultry growth and feather development. The additive is composed of scallop skirt oligopeptide and silkworm polypeptide in a 1:1 mass ratio. Compared with traditional chemical additives, the present invention has significant advantages: First, in terms of promoting poultry growth, the average daily weight gain of broilers treated with the combined bioadditive significantly increased by 9.43%, demonstrating a positive growth-promoting effect. More importantly, the invention demonstrated a remarkable synergistic effect on feather development. Experimental results showed that the length of the primary wing feathers of broilers in the combined bioadditive group increased significantly by 24.30%, with a synergistic effect index (q) value as high as 1.63, far exceeding the effects of either single group or high-dose addition alone.

[0024] Secondly, the present invention deeply explored the mechanism of action and found that the biological additive can synergistically promote the proliferation of chicken hair follicle epidermal stem cells, with the proliferation rate increased by 78.46%, which is better than the commonly used cell proliferation factor EGF, thereby accelerating feather growth from the source.

[0025] Finally, this invention, based on natural bioactive substances, avoids chemical residues and drug resistance, aligning with the philosophy of green, safe, and sustainable farming. Therefore, this invention not only significantly improves the economic value and overall health of poultry, but also provides a new, efficient, and environmentally friendly solution for modern animal husbandry. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a comparison of the average length of primary wing feathers in different treatment groups; Figure 2 This is a comparison of the proliferation rates of hair follicle epidermal stem cells in different treatment groups; exist Figure 1 and Figure 2 In the table, * indicates P ≤ 0.05, ** indicates P ≤ 0.01, and *** indicates P ≤ 0.001. DETAILED DESCRIPTION

[0027] Scallop skirts, a byproduct of aquatic product processing, are rich in protein, amino acids, and various trace elements. In recent years, they have been widely used in animal feed to enhance their nutritional value and promote the growth and development of livestock and poultry. Current research focuses on the positive effects of scallop skirts on growth performance, immune function, and intestinal health in aquatic and livestock species. However, research on their potential in poultry farming, particularly in improving feather growth, remains relatively scarce, with no systematic literature reports available.

[0028] Currently, research on Bombyx batryticatus primarily focuses on its active ingredients and their applications in medicine, healthcare, and certain livestock and poultry breeding fields. Rich in protein, amino acids, and various bioactive substances, Bombyx batryticatus is widely used in traditional Chinese medicine for its sedative, anticonvulsant, antibacterial, and immune-boosting properties. In recent years, studies have also attempted to apply it to livestock production, such as improving animal immunity and promoting growth performance. However, despite Bombyx batryticatus's excellent nutritional profile and bioactivity, research on its effects on poultry feather growth remains relatively scarce, with no systematic literature examining its effects on feather development.

[0029] Therefore, the present invention attempts to prepare scallop skirt and silkworm into oligopeptides and polypeptides respectively, and apply them to the poultry breeding process in order to promote the development of poultry feathers.

[0030] Example 1 (1) After thoroughly cleaning the scallop skirt, place it in boiling water and blanch for 2 minutes. After removing it, immediately place it in ice water to quickly cool it down and soak it for 30 minutes to prevent residual heat from adversely affecting the quality of the raw materials; (2) After draining, freeze-dry at -40°C, and then grind the dried scallop skirt into powder using a grinder, and pass through a 100-mesh sieve to obtain scallop skirt powder; (3) Deionized water was added to the scallop skirt powder at a solid-liquid ratio of 1:10, and the mixture was stirred and extracted at 4°C for 12 hours to obtain a scallop skirt aqueous extract; (4) The scallop skirt water extract was transferred to a constant temperature water bath, the temperature was adjusted to 55°C, the pH was 7.5, and a composite protease in an amount of 0.03 times the mass of the scallop skirt powder was added for enzymatic hydrolysis for 5 hours to obtain a scallop skirt enzymatic solution, wherein the composite protease consisted of flavor protease, alkaline protease and trypsin in a mass ratio of 5:3:2; (5) The enzymatic hydrolysate was heated to 95°C and maintained for 15 minutes to completely inactivate the enzyme. After cooling to room temperature, the solution was centrifuged at 12,000 rpm for 30 minutes at 4°C. The supernatant was collected to obtain the scallop skirt peptide solution. (6) After removing large particles of impurities from the scallop skirt peptide solution using a 0.45 μm filter membrane, the solution was filtered using 100 kDa, 10 kDa, and 1 kDa ultrafiltration membranes in sequence, retaining the permeate from each level and finally collecting the 1 kDa permeate; (7) The 1 kDa permeate was placed in a vacuum freeze dryer and freeze-dried to obtain scallop skirt oligopeptide.

[0031] Example 2 (1) After the dried silkworm pupae are cleaned with deionized water, the silkworm pupae are placed in liquid nitrogen for pre-freezing and hardening treatment to obtain pre-frozen silkworm pupae; (2) Add the pre-frozen silkworms into a mortar pre-cooled with liquid nitrogen, and grind them while continuously adding liquid nitrogen to maintain a low temperature environment until the silkworms are ground into powder; (3) Add petroleum ether in a powder:petroleum ether mass volume ratio of 1:3, stir at room temperature for 2 hours, centrifuge at 5000 rpm for 20 minutes, and collect the precipitate; (4) Deionized water was added at a solid-liquid ratio of 1:10 and stirred at 4°C overnight to obtain the silkworm extract; (5) The Bombyx batryticatus aqueous extract was transferred to a constant temperature water bath, the temperature was adjusted to 55°C, the pH was 7.5, and a composite protease in an amount of 0.03 times the mass of the Bombyx batryticatus powder was added for enzymatic hydrolysis for 5 hours to obtain a Bombyx batryticatus enzymatic hydrolyzate. The composite protease consisted of flavor protease, alkaline protease, and trypsin in a mass ratio of 5:3:2; (6) The enzymatic hydrolysate was heated to 95°C and maintained for 15 minutes to completely inactivate the enzyme. After cooling to room temperature, the solution was centrifuged at 12,000 rpm for 30 minutes at 4°C. The supernatant was collected to obtain the crude Bombyx batryticatus polypeptide solution.

[0032] (7) After filtering with a 0.45 μm microporous filter membrane, a Sephadex G-50 gel chromatography column with a column height of 40 cm and a diameter of 2 cm was used. Deionized water was used as the mobile phase and eluted at a flow rate of 0.5 mL / min. The eluate was collected in sections of 2 mL per tube. After all the elution was completed, all the eluate was collected and the eluate corresponding to the 280 nm elution peak was selected and combined to obtain the Bombyx batryticatus polypeptide solution; (8) The Bombyx batryticatus polypeptide solution is placed in a vacuum freeze dryer and freeze-dried to obtain the Bombyx batryticatus polypeptide.

[0033] Example 3 Animal experiments were conducted to test the effects of the scallop skirt oligopeptide and silkworm polypeptide on broiler growth and feather development (1) 450 AA broiler chickens (roosters) with an initial body weight of approximately 45 g were randomly divided into 5 groups, with 6 replicates in each group and 15 chickens in each replicate; The specific groupings are as follows: Control group: fed with basic feed, the basic feed selected was Zhengda broiler chicken compound feed 510, which provided a standard control for the experiment; Low-dose scallop skirt group: fed with basal feed + scallop skirt oligopeptide (30 mg of scallop skirt oligopeptide per 1 kg of basal feed), used to study the effects of low-dose scallop skirt oligopeptide alone on broiler growth and feather development; Low-dose Bombyx batryticatus polypeptide group: fed with basal feed + Bombyx batryticatus polypeptide (30 mg Bombyx batryticatus polypeptide was added to every 1 kg of basal feed), used to study the effects of low-dose Bombyx batryticatus polypeptide alone on broiler growth and feather development; Compound supplementation group: fed with basic feed + scallop skirt oligopeptide + silkworm polypeptide (30 mg scallop skirt oligopeptide and 30 mg silkworm polypeptide were added to each 1 kg of basic feed), to study whether the two polypeptides have a synergistic promoting effect when used in combination; High-dose scallop skirt group: fed with basal feed + scallop skirt oligopeptide (60 mg scallop skirt oligopeptide per 1 kg basal feed), used to study the effect of high-dose scallop skirt oligopeptide on the growth performance of broiler chickens; High-dose Bombyx batryticatus polypeptide group: fed with basal feed + Bombyx batryticatus polypeptide (60 mg Bombyx batryticatus polypeptide was added to every 1 kg of basal feed), used to study the effect of high-dose Bombyx batryticatus polypeptide on the growth performance of broiler chickens; Feeding method: Adopt free-feeding mode to ensure that broilers have access to sufficient feed at all times to meet their growth needs. Daily trough cleaning operation is carried out to promptly remove excess feed to prevent mold and contamination, and to ensure the freshness and hygiene of feed.

[0034] Drinking Water Management: Broilers were supplied with drinking water using nipple drinkers, with fresh water replaced daily to ensure clean and sanitary water sources, reduce the risk of disease associated with drinking water problems, and provide excellent drinking conditions for the healthy growth of broilers. The entire feeding and management process was strictly adhered to the AA Broiler Management Manual. From controlling environmental conditions such as temperature, humidity, and ventilation to daily hygiene and epidemic prevention, all procedures were followed according to standard procedures to minimize any interference with experimental results due to differences in feeding and management.

[0035] Routine vaccination treatment: On the first day, Marek's disease vaccine is administered subcutaneously; on the 7th day, Newcastle disease + infectious bronchitis combined live vaccine is administered through nasal / eye drops; on the 14th day, Fabricius bursal vaccine is administered through drinking water; on the 21st day, Newcastle disease (La Sota strain) vaccine is administered through drinking water; on the 28th day, avian influenza (H5+H7) vaccine is injected subcutaneously in the neck.

[0036] The experimental period was set to 42 days. At the beginning of the experiment (broiler chickens, 1 day old), the weight of each group of chickens was accurately recorded as initial data. At the end of the experiment (42 days old), the weight of each group of chickens was again recorded. By comparing the weight data at 1 and 42 days of age, growth performance indicators such as average daily weight gain were calculated to test the effects of the scallop skirt oligopeptides and silkworm polypeptides of the present invention on broiler growth.

[0037] The results are shown in Table 1: Table 1 Effects of the scallop skirt oligopeptide and silkworm polypeptide of the present invention on broiler growth

[0038] From the experimental results in Table 1, it can be seen that compared with the broiler control group fed only with the basic feed, the addition of scallop skirt oligopeptide, silkworm polypeptide, and the combined addition of the two to the feed promoted the growth rate of broilers to varying degrees, indicating that the scallop skirt oligopeptide and silkworm polypeptide prepared by the present invention have the potential to promote the growth performance of broilers to a certain extent.

[0039] Further comparison of the effects of each experimental group revealed that the group that added a combination of scallop skirt oligopeptides and silkworm polypeptides increased its growth rate by 9.43% compared to the control group. However, this increase was only slightly higher than the sum of the average daily weight gain of the low-dose scallop skirt oligopeptides group and the low-dose silkworm polypeptide group, and did not show a significant synergistic enhancement effect. In addition, the growth-promoting effect of the combined addition group was not statistically significant compared to the experimental group that used high-dose scallop skirt oligopeptides or high-dose silkworm polypeptides alone. Based on the above analysis, it can be inferred that when scallop skirt oligopeptides and silkworm polypeptides are used as feed additives in broiler farming, the two mainly exhibit an additive effect in terms of promotion.

[0040] Example 4 (1) After weighing at 42 days of age, 5 broilers were randomly selected from each replicate and sampled after slaughter; (2) Select the right wing of each chicken, locate the five longest and most fully developed primary wing feathers on the right wing, clamp them 1 cm above the feather root with hemostatic forceps, pull them out vertically along the direction of feather growth, and place them in a numbered sample bag; (3) After cleaning the blood stains at the base with saline, spread the feather flat on filter paper and place it in a constant humidity chamber (25°C, RH 50%) for 24 hours; (4) Use a vernier caliper to measure the length of the primary wing feathers and calculate the average primary wing feather length of each treatment group.

[0041] The results are shown in Table 2 and Figure 1 shown.

[0042] Table 2 Average length of primary wing feathers

[0043] As can be seen from the data in Table 2, compared with the control group, the average length of the primary wing feathers of broiler chickens in the low-dose scallop skirt oligopeptide group and the low-dose silkworm polypeptide group increased, with the increase in the low-dose silkworm polypeptide group being more significant. This result indicates that the scallop skirt oligopeptide and silkworm polypeptide prepared by the present invention both have the effect of promoting broiler feather growth to a certain extent. However, from the overall effect, the promotion effect of the two polypeptides on feather growth when added alone is still relatively limited.

[0044] Further comparison revealed that low-dose co-administration of scallop skirt oligopeptides and Bombyx batryticatus polypeptides increased primary wing feather length by 24.30%, significantly exceeding the effect of either single-dose combination. To verify the existence of a synergistic effect between the two, the synergistic effect calculation formula (q = E(AB) / (E(A) + E(B) - E(A) × E(B)) was used, yielding a q value of approximately 1.63. Since q≈1.63>1.15, this indicates that scallop skirt oligopeptides and Bombyx batryticatus polypeptides exhibit significant synergistic effects in promoting feather growth (primary wing feather growth).

[0045] Furthermore, at the same dosage, the combined supplementation group achieved a feather growth-promoting effect 2.08 times greater than that of the high-dose scallop skirt oligopeptide group and 1.67 times greater than that of the high-dose silkworm polypeptide group. This result further confirms the nonlinear enhancement effect of the combined use of the two, which is not only superior to the effect of each alone, but also superior to the effect of high-dose single addition.

[0046] Example 5 Obtaining AA broiler chicken hair follicle epidermal stem cells (1) Collect newly grown feathers (wing area) after plucking and quickly disinfect the feather base and surrounding skin with 75% alcohol; (2) Gently remove the feather with the hair follicle using tweezers, and rinse the hair follicle tissue three times in PBS containing 1% double antibody to remove floating debris and contamination; (3) After preliminary cleaning, the hair follicle tissue was transferred to the clean bench in the cell room and rinsed again with PBS containing 2% double antibody; (4) Remove the medulla and sheath of the hair follicle, cut the remaining part into pieces (less than 1 mm³) using sterile scissors, and transfer it to the digestion bottle; (5) Add 0.2% collagenase I solution and place in a 37°C water bath for digestion for 2 hours. After digestion is complete, add an equal volume of culture medium containing 10% FBS to terminate digestion; (6) Pass the digestion solution through a 40 μm cell sieve to remove large tissue pieces, and collect the cells by centrifugation at 1300 rpm for 5 minutes; (7) Wash once with PBS and resuspend the cells in DMEM complete medium. Inoculate the cells into a culture dish and place it in a 37°C, 5% CO2, saturated humidity incubator. (8) After 20 minutes, the culture medium was aspirated and inoculated into the culture dish of cells to remove non-stem cells such as fibroblasts with strong adherence; (9) After the cells grown to 90% confluence were digested, a portion of the cells were inoculated into a 96-well plate and fluorescently stained for CD29 (β1-integrin) and CD49f (α6-integrin) to identify the cells; (10) The remaining cells were subcultured. When CD29 and CD49f were positively expressed and located on the cell membrane, it indicated that the cells were hair follicle epidermal stem cells and were then used for subsequent experiments.

[0047] Example 6 From the perspective of cellular mechanism, it is detected whether the scallop skirt oligopeptide and silkworm polypeptide of the present invention can synergistically promote the proliferation of hair follicle epidermal stem cells Experimental preparation: Scallop skirt oligopeptide and silkworm polypeptide were prepared into 1 mg / ml stock solution using serum-free DMEM / F12 medium; Then, the scallop skirt oligopeptide mother solution and the silkworm polypeptide are prepared into a scallop skirt oligopeptide solution with a working concentration of 10 μg / mL, a 10 μg / mL silkworm polypeptide solution, and a composite solution containing a scallop skirt oligopeptide and a silkworm polypeptide with a working concentration of 10 μg / mL; At the same time, 10 ng / mL EGF solution was prepared as a positive control; Add 10% FBS before use.

[0048] Experimental steps: (1) The hair follicle epidermal stem cells obtained in Example 5 were seeded into 96-well collagen-coated plates, with 100 μL per well and a cell volume of 3×10 3 indivual; (2) After culturing the 96-well plate in a cell culture incubator for 24 hours, the cells were grouped as follows:

[0049] (3) After adding culture medium according to grouping, place in cell culture incubator and continue to culture for 48 hours; (4) At 0h and 48h, 10μL CCK-8 solution was added to each well, incubated at 37℃ in the dark for 2h, and the absorbance at 450nm was measured by microplate reader to calculate the proliferation rate: proliferation rate (%) = OD value of experimental group at 48h 450 − 0hOD 450 ×100% / control group 48hOD 450 −0h OD 450 The results are shown in Table 3 and Figure 2 shown.

[0050] Table 3 Differences in absorbance and proliferation rate among different groups

[0051] From the experimental results of Table 3 and Figure 2 It can be seen that, compared with the control group, the addition of scallop skirt oligopeptide, polypeptide of jieshuan, the combination of the two and the EGF positive control can significantly promote the proliferation ability of chicken hair follicle epidermal stem cells, indicating that these treatment factors can promote the growth of chicken hair follicle epidermal stem cells to a certain extent.

[0052] Further comparison of the effects of each treatment group found that the combined addition group (i.e. scallop skirt oligopeptide and jieshuan polypeptide were used together) had the most significant promoting effect on the proliferation of chicken hair follicle epidermal stem cells. Compared with the control group, its proliferation rate increased by 78.46%, which was not only significantly higher than that of the single addition group, but also exceeded the effect of the commonly used cell proliferation factor EGF positive control group, showing the superiority of the compound formula in the functional regulation of chicken hair follicle epidermal stem cells.

[0053] Further calculation of q value can see that q = E(AB) / (E(A) + E(B) - E(A)×E(B))≈1.54, which shows that the scallop skirt oligopeptide and jieshuan polypeptide of the present application can synergistically promote the proliferation of chicken hair follicle epidermal stem cells when used together, thereby helping to achieve the effect of promoting chicken feather growth.

[0054] Example 7 A cell culture preparation for promoting the proliferation of poultry hair follicle epidermal stem cells The biological preparation is composed of DMEM / F12 culture medium, 10% fetal bovine serum, 10 μg / mL scallop skirt oligopeptide, 10 μg / mL jieshuan polypeptide; The cell culture preparation is used for chicken hair follicle epidermal stem cell proliferation promoting culture.

Claims

1. A biological additive for promoting poultry growth and feather development, characterized in that: The biological additive consists of scallop skirt oligopeptide and silkworm polypeptide, and the mass ratio of the scallop skirt oligopeptide to the silkworm polypeptide is 1:

1.

2. The biological additive according to claim 1, characterized in that The preparation method of the scallop skirt oligopeptide comprises the following steps: (1) After cleaning the scallop skirt, blanch it in boiling water, cool it in ice water and drain it; (2) freeze-drying and then crushing and sieving to obtain scallop skirt powder; (3) Deionized water was added to the scallop skirt powder at a solid-liquid ratio of 1:10, and the mixture was extracted at 4°C for 12 hours to obtain a scallop skirt aqueous extract; (4) adjusting the pH of the scallop skirt water extract to 7.5 and 55° C., adding a composite protease for enzymatic hydrolysis for 5 hours, wherein the composite protease comprises flavor protease, alkaline protease, and trypsin in a mass ratio of 5:3:2; (5) After inactivation and centrifugation, the enzymatic hydrolysate was filtered using 100 kDa, 10 kDa, and 1 kDa ultrafiltration membranes, and the 1 kDa permeate was collected; (6) The 1 kDa permeate was freeze-dried to obtain scallop skirt oligopeptide.

3. The biological additive according to claim 2, wherein the preparation method of the silkworm polypeptide comprises the following steps: (a) Bombyx batryticatus was pre-frozen in liquid nitrogen, ground into powder, and defatted with petroleum ether. (b) Deionized water was added at a solid-to-liquid ratio of 1:10 and extracted overnight at 4°C to obtain a water extract; (c) adjusting the aqueous extract to pH 7.5 and 55° C., and adding a composite protease for enzymatic hydrolysis for 5 hours, wherein the composite protease comprises flavor protease, alkaline protease, and trypsin in a mass ratio of 5:3:2; (d) The enzymatic hydrolysate was inactivated and centrifuged, and then purified by Sephadex G-50 gel chromatography column. The fractions corresponding to the absorption peak at 280 nm were collected and lyophilized to obtain Bombyx batryticatus polypeptide.

4. The biological additive according to claim 1, characterized in that The amount of the biological additive used when added to feed is 60 mg per 1 kg of basic feed; The biological additive achieves the effect of promoting poultry feather development by promoting the proliferation of poultry hair follicle epidermal stem cells.

5. Use of a biological additive composed of scallop skirt oligopeptide and silkworm polypeptide in preparing feed for synergistically promoting poultry feather development, characterized in that: The scallop skirt oligopeptide is prepared by the preparation method described in claim 2; The Bombyx batryticatus polypeptide is prepared by the preparation method described in claim 3; In the biological additive, the mass ratio of the scallop skirt oligopeptide to the silkworm polypeptide is 1:

1.

6. The use according to claim 5, characterized in that The feed consists of basic feed and biological additives; In the feed, the usage amount of the biological additive corresponding to each 1 kg of the basic feed is 60 mg.

7. A feed for promoting poultry growth and feather development, characterized in that: The feed is composed of a basic feed and the biological additive according to any one of claims 1 to 4; In the feed, the usage amount of the biological additive corresponding to each 1 kg of the basic feed is 60 mg.

8. Use of a biological additive composed of scallop skirt oligopeptide and silkworm polypeptide in the preparation of a cell culture preparation for synergistically promoting the proliferation of poultry hair follicle epidermal stem cells, characterized in that: The scallop skirt oligopeptide is prepared by the preparation method described in claim 2; The Bombyx batryticatus polypeptide is prepared by the preparation method described in claim 3; In the biological additive, the mass ratio of the scallop skirt oligopeptide to the silkworm polypeptide is 1:

1.

9. The use according to claim 8, characterized in that The biological preparation consists of biological additives, DMEM basal culture medium and 10% fetal bovine serum; In the biological preparation, the added concentration of the biological additive is 20 μg / mL.

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