Poultry feed additive containing nuciferine and application thereof

Abstract

The invention discloses a poultry feed additive containing nuciferine and application thereof. The invention claims to protect nuciferine or application of the poultry feed additive in preparation of poultry feed. The application comprises any one or more of 1) to 6): 1) improving the growth performance of laying hens; (2) the quality of eggs is improved; (3) the immune performance of laying hens is improved; 4) the oxidation resistance of the laying hens is improved; (5) the lipid metabolism capability of the laying hens is improved; and 6) increasing the abundance of short-chain fatty acids in cecum contents of laying hens. The invention has the following beneficial effects: by supplementing nuciferine with daily ration, the egg quality of the Jiangshan black-bone chicken can be significantly improved, the immune performance and antioxidant ability of the Jiangshan black-bone chicken in the late laying period can be improved, inflammatory response can be reduced, lipid metabolism can be improved, and the abundance of cecum short-chain fatty acid can be improved; nuciferine is used as a potential poultry feed antibiotic substitute, and plays an important role in nutrition regulation and control of Jiangshan black-bone chickens in the later egg laying period.

Description

Technical Field

[0001] The present application belongs to the field of poultry feed, and relates to a poultry feed additive containing nuciferine and application thereof. Background Art

[0002] Chicken is an important poultry, a major source of both meat and eggs, and an important animal model due to its small size and anatomical similarity to most birds. In poultry, nutrient digestion and absorption mainly occur in the crop, gizzard, and small intestine, which plays an important role in the production performance and health of chickens. Jiangshan Black-bone Chicken is a local fine breed for both egg and meat. Its origin and central production areas are Jiangshan Tanshi, Shuangta, Hushan, Qinghu and other townships. It has white feathers, black skin, black crown, and black toes. It is a famous high-quality special black-bone chicken variety in my country.

[0003] The intestine is the main site for digestion and absorption of animal nutrients, and its healthy development is crucial to poultry growth. In the late egg-laying period, laying hens have an imbalance of intestinal flora, disordered metabolic functions, and weakened digestion and absorption of nutrients, which will lead to a significant decline in growth performance, seriously restricting the development of the poultry industry. Therefore, improving the intestinal health of poultry is of great significance to its production. At present, the application scale of plant-based feed alternatives is growing, especially in Europe, where some natural plants such as oregano and thyme have been developed into alternatives for many years due to the early ban on antibiotics in feed. The main functional active ingredients of plants are secondary metabolites synthesized during the natural growth of plants, such as alkaloids, polyphenols, volatile oils, flavonoids, plant polysaccharides, organic acids, saponins, etc. These active ingredients have positive effects in improving animal health, preventing animal diseases, and promoting animal growth, and can be used as substitutes for feed antibiotics in animal production. Currently, most of the plant-based feed additives for laying hens have complex formulas. For example, Chinese Patent 201410670441.2 discloses a plant-based feed additive for laying hens. However, the patent studies the addition of Moringa leaf powder to the feed for feeding chicks, and found that their feed intake, total weight gain, average daily weight gain and feed conversion rate were significantly improved. On the one hand, no research was conducted on laying hens in the late egg-laying period, and there is a lack of corresponding data, so its feasibility remains to be verified. On the other hand, the plant-based feed additive has complex ingredients, is inconvenient to prepare, has a high production cost, and is not easy to industrialize.

[0004] Lotus (Nelumbo Nucifera Gaertn.) is a plant of the family Nelumbo that is widely cultivated in Asia and Africa. Lotus is a valuable medical resource among the people. Lotus leaves have been considered to have anti-obesity effects since the Ming Dynasty. The earliest medical book "Essentials of Syndrome Differentiation and Treatment" pointed out that lotus leaves can help lose weight. In recent years, lotus leaves have also become popular in China as tea beverages and dietary supplements for weight loss and lipid reduction. Lotus leaves are rich in a variety of biologically active ingredients, such as lotus leaf flavonoids, lotus leaf alkaloids, lotus leaf essential oils and lotus leaf polysaccharides, among which lotus leaf flavonoids and nuciferine are the main active substances. Nuciferine (NF) is a major aporphine alkaloid component in lotus leaves, which has been shown to have anti-obesity and antioxidant properties. Another report pointed out that nuciferine inhibits the expression of p-AKT, PI3K and IL-1β in a dose-dependent manner, thereby showing anti-tumor activity and reducing the risk of colorectal cancer. Lotus leaf flavonoids have the effects of lowering blood lipids, treating and preventing allergies, cancer, cardiovascular diseases, as well as anti-aging, antibacterial and antioxidant effects. Although some people have developed a feed additive suitable for pigs and sows, as proposed in Patent 201510493152.4, which contains lotus leaf alkaloids, the application is aimed at pigs and sows, and lotus leaf alkaloids are added to the feed additive in the form of a composition, with the purpose of increasing the content of nutrients such as crude protein in pork, increasing the edible value of pork, increasing the litter birth rate of sows, and improving the production performance of sows, and these performance improvements do not show a direct relationship with the above-mentioned performance improvements.

[0005] In summary, the current research on nuciferine is mainly focused on its medicinal value and its application as a drug in the treatment of certain diseases. There is no report on the effect of adding nuciferine to the diet of laying hens, especially laying hens in the late laying period. Summary of the invention

[0006] In view of the shortcomings of the prior art mentioned above, the purpose of the present application is to provide a poultry feed additive containing nuciferine and its application, so as to enhance immune performance and antioxidant capacity, reduce inflammatory response, improve lipid metabolism, reshape the abundance of short-chain fatty acids in the cecum, and optimize the flora structure.

[0007] To solve the above problems, the technical solution adopted in this application is:

[0008] The invention provides a poultry feed additive with nuciferine as an effective ingredient.

[0009] The invention also provides application of nuciferine or the poultry feed additive in preparing poultry feed.

[0010] As a preferred embodiment of the present invention, the poultry feed is chicken feed for feeding laying hens.

[0011] As a preferred embodiment of the present invention, the application includes any one or more of 1) to 6):

[0012] 1) Improve the production performance of laying hens;

[0013] 2) Improve egg quality in eggs;

[0014] 3) Improve the immune performance of laying hens;

[0015] 4) Improve the antioxidant properties of laying hens;

[0016] 5) Improve the lipid metabolism capacity of laying hens;

[0017] 6) Increase the abundance of short-chain fatty acids in the cecal contents of laying hens.

[0018] As a preferred embodiment of the present invention, the application 1) of improving the production performance of laying hens includes:

[0019] Increase feed intake of laying hens;

[0020] Increase egg weight;

[0021] As a preferred embodiment of the present invention, the method for improving the egg quality of laying hens in application 2) comprises:

[0022] Increase the albumen height of eggs; and / or

[0023] Increase the Haugh unit of the egg.

[0024] As a preferred embodiment of the present invention, the method of improving the immune performance and antioxidant performance of laying hens in application 3) includes:

[0025] Increase the content of anti-inflammatory factors in the serum of laying hens;

[0026] Reducing the levels of pro-inflammatory factors in the serum of laying hens; and / or

[0027] Increase the levels of immunoglobulins IgA, IgG, and IgM in serum.

[0028] As a preferred embodiment of the present invention, the method for improving the antioxidant performance of laying hens in application 4) comprises:

[0029] Improving the total antioxidant capacity, catalase and glutathione peroxidase content in the liver and / or serum of laying hens; and / or

[0030] Reduction of malondialdehyde content in the liver and / or serum of laying hens.

[0031] As a preferred embodiment of the present invention, the method of improving the lipid metabolism capacity of laying hens in application 5) comprises:

[0032] Increase the levels of T-CHO and AST in serum.

[0033] As a preferred embodiment of the present invention, the method of increasing the abundance of short-chain fatty acids in the cecal contents of laying hens in application 6) comprises:

[0034] Increase the content of propionic acid, butyric acid and valeric acid in the cecal contents.

[0035] As a preferred embodiment of the present invention, the application comprises the following steps:

[0036] Nuciferine or the poultry feed additive is added into the basic diet of laying hens.

[0037] As a preferred embodiment of the present invention, the amount of nuciferine added to the basal diet is 200-600 mg / kg.

[0038] As a preferred embodiment of the present invention, the amount of nuciferine added to the basal diet is 200 mg / kg, 400 mg / kg or 600 mg / kg.

[0039] As a preferred embodiment of the present invention, the laying hens are laying hens that are 400 days old and above.

[0040] As a preferred embodiment of the present invention, the breed of laying hen is Jiangshan black-bone chicken.

[0041] The present invention also provides a feeding method of poultry feed, characterized in that: the poultry feed containing nuciferine or the poultry feed additive is used as the daily diet of laying hens, and the laying hens are fed twice a day, with the time interval between the two feedings being not less than 6 hours, and the chickens are ensured to have free drinking water and 16 hours of light per day during the feeding period, and the laying hens are raised on the ground with a stocking density of less than 4 chickens / m2, and are provided with a suitable exercise area.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] (1) Dietary supplementation of nelumboide. When the amount of nelumboide added to the basic diet is 200-600 mg / kg, it can significantly improve the growth performance of laying hens, especially in the late egg-laying period, and the egg quality. When 400 mg / kg of nelumboide is added to the diet, the average egg weight can be significantly increased. When 600 mg / kg of nelumboide is added, the feed intake, egg-laying feed-egg ratio, and egg albumin height and Haugh unit of laying hens can be significantly increased, and it will not affect the eggshell strength, egg shape index, and yolk color.

[0044] (2) Improve the immune function of laying hens, reduce inflammatory response, and improve lipid metabolism. When lotus leaf alkaloids are added to the diet, the pro-inflammatory factors in the serum are significantly reduced, the anti-inflammatory factors are significantly increased, and the immunoglobulins IgA, IgG, and IgM in the serum are significantly increased;

[0045] (3) Improving the antioxidant performance of laying hens. Adding lotus leaf alkaloids to the diet can significantly increase the content of T-AOC in the liver and serum, and significantly reduce the content of MDA in the liver; it can also significantly increase the content of T-AOC, CAT and GSH-Px in the serum. Since T-AOC, MDA, GSH-Px and CAT in serum are related to antioxidant activity, increasing the positively correlated T-AOC, GSH-Px and CAT in serum and reducing the negatively correlated MDA content in serum can significantly improve the antioxidant performance of laying hens.

[0046] (4) Improve the lipid metabolism performance of laying hens. Adding nuciferine to the diet can significantly increase the content of T-CHO and AST in serum. T-CHO, TG, HDL-C, and LDL-C in serum are related to lipid metabolism. Therefore, by increasing the content of T-CHO and AST in serum, the content of T-CHO and AST in serum can be increased.

[0047] (5) Increase the abundance of short-chain fatty acids in the cecum. In particular, adding nuciferine to the diet can significantly increase the content of propionic acid, butyric acid, and valeric acid in the cecum of laying hens. Then, by scientifically controlling the abundance of short-chain fatty acids, the intestinal health of laying hens can be improved.

[0048] (6) As a potential substitute for antibiotics in poultry feed, nuciferine plays an important role in the nutritional regulation of laying hens, especially Jiangshan black-bone chickens. DETAILED DESCRIPTION

[0049] The following describes the implementation of the present application through specific examples, and those skilled in the art can easily understand other advantages and effects of the present application from the contents disclosed in this specification. The present application can also be implemented or applied through other different specific implementations, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present application.

[0050] It should be noted that the process equipment or devices not specifically specified in the following embodiments are all conventional equipment or devices in the art.

[0051] In addition, it should be understood that one or more method steps mentioned in this application do not exclude the existence of other method steps before or after the combination step or the insertion of other method steps between these explicitly mentioned steps, unless otherwise specified; it should also be understood that the combination connection relationship between one or more devices / apparatuses mentioned in this application does not exclude the existence of other devices / apparatuses before or after the combination device / apparatus or the insertion of other devices / apparatuses between these two explicitly mentioned devices / apparatuses, unless otherwise specified. Moreover, unless otherwise specified, the numbering of each method step is only a convenient tool for identifying each method step, and is not intended to limit the order of arrangement of each method step or the scope of implementation of this application. Changes or adjustments in their relative relationships shall also be regarded as the scope of implementation of this application without substantial changes in the technical content.

[0052] In the following examples of the present application, the nuciferine used is a plant extract nuciferine purchased from Shaanxi Feimi Biotechnology Co., Ltd.

[0053] The present application is further described below in conjunction with specific embodiments, but the protection scope of the present application is not limited thereto.

[0054] Example 1

[0055] 1. Animal testing and diet

[0056] The animal experiment was carried out in Jiangshan Lanfeng Poultry Breeding Co., Ltd. 319 healthy Jiangshan black-bone chickens aged 400 days were selected. They were randomly divided into 5 groups, each with 6 replicates (including 3 replicates in the antibiotic group), and 70 chickens in each group (including 74 in the negative group). The grouping and the amount of lotus leaf alkaloids added in each group are shown in Table 1. The basic diet was prepared with reference to the recommended values ​​of the nutrient requirements of laying hens in NRC1994 "Nutritional Requirements of Poultry". The feeding cycle was 7 weeks, with one week of pre-test and 6 weeks of formal experiment. Feed twice a day (6:30, 15:30) to ensure free drinking water for the flock. During the experiment, the light was 16 hours a day, and the chickens were raised on the ground. The stocking density was controlled at a low level (<4 / m2), and a suitable exercise area was provided. The feed intake, egg production, and egg weight were recorded every day.

[0057] Table 1 Experimental groups

[0058]

[0059]

[0060] 2. Sample collection and index testing

[0061] 2.1 Sample collection

[0062] One day before the end of the experiment, the feed was stopped 8 hours in advance. Two experimental chickens with a body weight close to the average body weight were randomly selected from each group and each replicate for wing vein blood collection. After standing for 30 minutes, the serum was collected by centrifugation at 4°C and 3000r / min and frozen at -20°C for later use. After blood collection, the experimental chickens were slaughtered, and their cecal contents were collected in cryopreservation tubes and stored in a -80°C refrigerator.

[0063] 2.2 Index determination

[0064] 2.2.1 Production performance measurement

[0065] The number of eggs laid, broken eggs, egg weight and dead chickens were recorded daily, per group and per replicate, and the weekly feed consumption was calculated. The measured data were used to calculate daily feed intake, average egg weight, egg production rate and feed-egg ratio.

[0066] 2.2.2 Egg quality determination

[0067] 24 eggs were selected for egg quality determination in each replicate 3-5 days before sampling. Egg weight and yolk weight were weighed using an electronic balance (1 / 10,000, YH-M10001). Egg shape index was determined using a digital egg tester (DET-6000, NABEL, Japan), eggshell strength was measured using an eggshell strength tester, and albumen height, Haugh units (HU), and yolk color were measured using an egg quality tester. Eggshell thickness was measured using an eggshell thickness meter (Robotmation Co., Ltd., Japan), and the values ​​were averaged at three different locations on the top, middle, and bottom of the egg.

[0068] 2.2.3 Immunity performance test

[0069] The levels of pro-inflammatory factors in serum: tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), interleukin 6 (IL-6), and anti-inflammatory factors: interleukin 10 (IL-10) and transforming growth factor β (TGF-β), and immunoglobulins: IgA, IgG, and IgM were detected.

[0070] The specific test method is: use enzyme-linked immunosorbent assay kit for determination. The test kits were purchased from Wuhan Huamei Biological Co., Ltd., and the reagent preparation and operation steps were carried out according to the instructions.

[0071] 2.2.4 Antioxidant capacity test

[0072] Serum antioxidant indicators: total antioxidant capacity (T-AOC), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and catalase (CAT) activity; liver antioxidant: total antioxidant capacity (T-AOC), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and catalase (CAT) activity.

[0073] The specific test method is: use an ELISA instrument for determination, the test kits are purchased from Nanjing Jiancheng Bioengineering Institute, and the reagent preparation and operation steps are carried out according to the instructions.

[0074] 2.2.5 Serum biochemical indexes

[0075] The levels of total protein (TP), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bile acid (TBA), total cholesterol (T-CHO), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in serum were tested.

[0076] The specific test method is: use Roche automatic biochemical analyzer for determination, the test kits are purchased from Shanghai Roche Company, and the reagent preparation and operation steps are carried out according to the instructions.

[0077] 2.2.6 Detection of short-chain fatty acids in cecal contents

[0078] The content of acetic acid, propionic acid, butyric acid, valeric acid, isobutyric acid and isovaleric acid in the cecal contents of laying hens was determined by gas chromatography-mass spectrometry (GC-MS). The specific detection method is: take 0.1g of cecal contents, add 500μL ultrapure water to homogenize, and centrifuge at 4℃ and 12000r / min for 5min. Then take 200μL of supernatant, add 0.1gNaCl, 20μL 50% concentrated sulfuric acid, 200μL anhydrous ether and 8μL internal standard to fully homogenize, centrifuge again at 4℃ and 12000r / min for 5min, take 100μL supernatant, and put it on GC-MS for testing.

[0079] 3. Data Analysis

[0080] The data are presented in the form of "mean ± standard error" for each group of samples. All data were analyzed using SPSS software, and the P value was used to determine whether the results were significant. Statistically, a P value less than 0.05 is considered a significant difference, a P value between 0.05-0.1 indicates a statistical trend, and a P value greater than 0.1 indicates no significant difference.

[0081] 4. Results

[0082] 4.1 Production performance

[0083] As shown in Table 2, adding nelumboline to the diet during the 1st to 3rd week of the experiment can significantly increase the average feed intake (P < 0.001). Adding 400 mg / kg nelumboline to the diet can significantly increase the average egg weight (P < 0.05). Compared with the negative control group, adding 600 mg / kg nelumboline can significantly increase the feed-egg ratio (P < 0.05). Adding nelumboline to the diet during the 4th to 6th week of the experiment can significantly increase the average feed intake (P < 0.001). Compared with the negative control group, adding 600 mg / kg nelumboline can significantly increase the feed-egg ratio (P < 0.05).

[0084] Table 2 Effect of nuciferine on growth performance of Jiangshan black-bone chicken

[0085]

[0086] 4.2 Egg quality

[0087] As shown in Table 3, adding nuciferine to the diet had no significant effect on the eggshell strength, egg shape index and yolk color of Jiangshan black-bone chickens (P>0.05). Compared with the negative control group, adding nuciferine to the diet can significantly increase the protein height and Haugh unit of Jiangshan black-bone chickens (P<0.05), among which the effect of adding 600 mg / kg nuciferine was the best.

[0088] Table 3 Effect of nuciferine on the quality of Jiangshan black-bone eggs

[0089]

[0090] 4.3 Cytokines

[0091] As shown in Table 4, compared with the antibiotic group, the addition of nuciferine to the diet significantly reduced the pro-inflammatory factors in the serum, and significantly increased the anti-inflammatory factors (P < 0.001). Among them, the addition of 600 mg / kg nuciferine had the best effect.

[0092] Table 4 Effects of nuciferine on pro-inflammatory and anti-inflammatory factors in Jiangshan black-bone chicken

[0093]

[0094] 4.4 Immune performance

[0095] As shown in Table 5, compared with the negative control group, the serum immunoglobulins IgA, IgG and IgM increased significantly when nuciferine was added to the diet (P < 0.001). The best effect was achieved when 200 mg / kg nuciferine was added.

[0096] Table 5 Effect of nuciferine on immunoglobulin in serum of Jiangshan black-bone chicken

[0097]

[0098] 4.5 Antioxidant properties

[0099] As shown in Table 6, compared with the antibiotic group, adding nuciferine to the diet can significantly increase the content of T-AOC in the liver (P < 0.001). Compared with the negative control group, adding nuciferine to the diet can significantly reduce the content of MDA in the liver (P < 0.001).

[0100] Table 6 Effect of nuciferine on antioxidant activity in Jiangshan black-bone chicken liver

[0101]

[0102] 4.6 Serum biochemical indexes

[0103] T-AOC, MDA, GSH-Px, and CAT in serum are related to antioxidant activity. As shown in Table 7, the addition of nelumboline to the diet can significantly reduce the content of MDA in serum, and the reduction effect of adding 200 mg / kg nelumboline is the best (P < 0.001). It can also significantly increase the content of T-AOC and CAT in serum (P < 0.001). Compared with the antibiotic group, nelumboline can significantly increase the content of GSH-Px in serum (P < 0.001). T-CHO, TG, HDL-C, and LDL-C in serum are related to lipid metabolism. The addition of nelumboline to the diet had no significant effect on the content of TG, HDL-C, and LDL-C in serum (P > 0.05). Compared with the negative control group, as the content of added nelumboline increased, the content of T-CHO in serum also gradually increased, and the addition of 600 mg / kg nelumboline significantly increased T-CHO (P < 0.05). ALT, AST, ALP, TBA, ALB, and TP in serum are related to liver function. Adding nuciferine to the diet had no significant effect on the levels of ALT, ALP, TBA, and TP in serum (P>0.05). Compared with the negative control group, adding nuciferine could significantly increase the content of AST in serum (P<0.05).

[0104] Table 7 Serum biochemical indexes of Jiangshan black-bone chicken

[0105]

[0106] 4.7 Short-chain fatty acids

[0107] As shown in Table 8, the addition of nuciferine to the diet had no significant effect on the content of isobutyric acid and isovaleric acid in the cecal contents (P>0.05). Compared with the antibiotic group, the addition of 200 mg / kg nuciferine significantly increased the content of propionic acid, butyric acid, and valeric acid in the cecal contents (P<0.05).

[0108] Table 8 Effect of nuciferine on short-chain fatty acids in cecal contents of Jiangshan black-bone chicken

[0109]

[0110] In summary, dietary supplementation of nuciferine can significantly improve the egg quality of Jiangshan black-bone chickens, enhance the immune performance and antioxidant capacity of Jiangshan black-bone chickens in the late egg-laying period, reduce inflammatory response, improve lipid metabolism, and reshape the abundance of cecal short-chain fatty acids. As a potential substitute for poultry feed antibiotics, nuciferine plays an important role in the nutritional regulation of Jiangshan black-bone chickens.

[0111] The above examples are for the purpose of illustrating the embodiments disclosed in the present application and should not be construed as limiting the present application. In addition, the various modifications listed herein and the variations of methods and compositions in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present application. Although the present application has been specifically described in conjunction with various specific preferred embodiments of the present application, it should be understood that the present application should not be limited to these specific embodiments. In fact, various modifications obvious to those skilled in the art as described above to obtain inventions should be included within the scope of the present application.

Claims

1. A poultry feed additive with nuciferine as an effective ingredient.

2. Use of nuciferine or the poultry feed additive according to claim 1 in preparing poultry feed.

3. The use according to claim 2, characterized in that: The poultry feed is chicken feed for feeding laying hens.

4. The use according to claim 3, characterized in that: The application includes any one or more of 1) to 6): 1) Improve the production performance of laying hens; 2) Improve egg quality in eggs; 3) Improve the immune performance of laying hens; 4) Improve the antioxidant properties of laying hens; 5) Improve the lipid metabolism capacity of laying hens; 6) Increase the abundance of short-chain fatty acids in the cecal contents of laying hens.

5. The use according to claim 4, characterized in that: The application 3) includes: Increase the content of anti-inflammatory factors in the serum of laying hens; Reducing the levels of pro-inflammatory factors in the serum of laying hens; and / or Increase the levels of immunoglobulins IgA, IgG, and IgM in serum.

6. The use according to claim 4, characterized in that: The applications all include the following steps: Nuciferine or the poultry feed additive is added into the basic diet of laying hens.

7. The use according to any one of claims 4 to 6, characterized in that: The amount of nuciferine added to the basic diet is 200-600 mg / kg.

8. The use according to claim 7, characterized in that: The amount of nuciferine added to the basic diet is 200 mg / kg, 400 mg / kg or 600 mg / kg.

9. The use according to claim 3, characterized in that: The laying hens are laying hens that are 400 days old or older.

10. A feeding method for poultry feed, characterized in that: The poultry feed containing nuciferine or the poultry feed additive according to claim 1 is used as the daily diet of laying hens, and the feeding is carried out twice a day, with an interval of not less than 6 hours between the two feedings. During the feeding period, the chickens are ensured to have free drinking water and 16 hours of light per day. The laying hens are raised on the ground with a stocking density of <4 / m 2 , and provide suitable sports venues.

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