An anti-immune stress composition for chickens and its preparation method and application

Abstract

The present invention discloses an anti-immune stress composition for chickens, a preparation method and an application thereof. The composition is composed of the following components in parts by weight: 15 to 25 parts of lotus leaves, 15 to 25 parts of Platycladus orientalis leaves, 10 to 20 parts of licorice polysaccharide, 0.1 to 0.5 parts of vitamin E, and 0.05 to 0.5 parts of nano zinc oxide or paeonol-capped nano zinc oxide. The anti-immune stress composition for chickens of the present invention, through the addition of traditional Chinese medicines with antioxidant, antibacterial, antiviral and liver protection properties, as well as nano zinc oxide and vitamin E, the components work synergistically, effectively improving the immune stress status of chickens, improving body metabolism, effectively improving the immunity of chickens, increasing the feed intake of chickens under immune stress, stimulating the digestion and absorption function of the gastrointestinal tract of chickens, and improving the ability of chickens to resist immune stress.

Description

Technical Field

[0001] The invention relates to an anti-immune stress composition for chickens and a preparation method and application thereof, belonging to the technical field of biological health care and feed additives. Background Art

[0002] Immunological stress, in a narrow sense, refers to the systemic nonspecific adaptive reaction of the body caused by the immune response of animals after being vaccinated with antigens. In modern intensive chicken production, vaccines play an extremely important role in preventing, controlling and eliminating poultry infectious diseases. On the one hand, vaccination protects the body from pathogens, and on the other hand, it also causes immune stress as a special stressor, which has certain harm to the body's tissue metabolism or immune system, not only affecting the growth and development and production performance of livestock and poultry, but may also cause the death of some animals. Especially in the chick stage, the immune system has not yet fully developed, but frequent immunization is carried out about once a week, which puts the body in a state of immune stress for a long time. Acute stress will enhance the body's immune response, while long-term chronic stress will inhibit the body's immune function, causing the chicken to grow slowly and reduce its disease resistance. Immune stress can cause animals to show lack of energy, lethargy, and decreased appetite during production. Its material metabolism will also change, causing the animals' needs to maintain growth to be converted into needs to maintain immune stress responses, thereby hindering the growth and development of livestock and poultry and reducing production performance, posing great potential harm to the breeding industry.

[0003] In a broad sense, "immune stress" is also called immune stimulation, which refers to the fact that animals are frequently attacked by various pathogenic microorganisms in a poor breeding environment or under sub-health conditions, which causes the immune system to be constantly activated, produces immune responses to resist microbial invasion, and causes animal trauma, infection, or internal tumors and other non-specific adaptive reactions. When viruses, bacteria, and endotoxins invade the body, the immune system is highly activated, promoting immune cells to secrete inflammatory cytokines, including IL-1, IL-2, and TNF-α, and regulating the synthesis of hormones such as adrenal hormones, glucocorticoids, and growth hormones, affecting the body's tissue metabolism and neuroendocrine system, directly or indirectly causing a series of changes in the body's behavior and material metabolism, and inhibiting the growth of livestock and poultry.

[0004] Therefore, various stress factors such as over-immunization, pathogen infection, drug abuse and poor sanitation of the breeding environment can induce immune stress in chickens. Under immune stress, chickens experience increased body temperature, loss of appetite, slow growth, and disordered intestinal microbial flora. Under long-term immune stress, the immune organ index of chickens decreases, immune function is suppressed, the level of cellular inflammatory factors (such as IL-1, IL-6, TNF-α, etc.) increases abnormally, resistance to disease decreases, and the body is in sub-healthy condition. As a result, the growth and development of livestock and poultry are hindered, production performance is reduced, the meat quality of broilers is affected, and great potential harm is caused to the poultry industry.

[0005] The Chinese invention patent application document with application publication number CN 109619286 A discloses "a broiler chicken anti-vaccine immune stress agent and its application". The broiler chicken anti-vaccine immune stress agent contains turmeric, astaxanthin, oligosaccharide, threonine, methionine, composite multidimensional and carrier, but the amount of its components is large and the price is expensive, which is not suitable for large-scale promotion. Therefore, the development of a chicken anti-immune stress product with simple ingredients, low price and good anti-immune stress effect has become an urgent problem to be solved in the modern poultry industry. Summary of the invention

[0006] In order to solve the above problems, the first purpose of the present invention is to provide an anti-immune stress composition for chickens. The composition of the present invention comprehensively prevents and treats the immune stress of chickens by targeting anti-inflammation, anti-oxidation, liver protection, intestinal protection and endocrine regulation, so as to improve the health and production performance of chickens and reduce the mortality rate.

[0007] The second object of the present invention is to provide a method for preparing an anti-immune stress composition for chickens.

[0008] The third object of the present invention is to provide the use of the anti-immune stress composition for chickens in preparing feed for enhancing the anti-immune stress ability of chickens.

[0009] In order to achieve the above object, the technical solution adopted by the present invention is:

[0010] An anti-immune stress composition for chickens is composed of the following components in parts by weight: 15-25 parts of lotus leaves, 15-25 parts of Platycladus orientalis leaves, 10-20 parts of licorice polysaccharide, 0.1-0.5 parts of vitamin E, and 0.05-0.5 parts of nano zinc oxide or paeonol-capped nano zinc oxide.

[0011] The anti-immune stress composition for chickens of the present invention can effectively alleviate diarrhea, constipation, decreased production performance, decreased quality of livestock products, etc. caused by intestinal flora imbalance, increased oxygen free radicals, liver and gallbladder damage, etc. caused by immune stress. The raw materials used are green and efficient, will not cause any side effects to animals, and will not affect environmental safety, has obvious economic and ecological benefits, and is easy to promote on a large scale in practical applications.

[0012] In the composition, the functions of each component are as follows:

[0013] Lotus leaf: It is the leaf of the lotus plant of the Nymphaeaceae family. Its chemical components mainly include nuciferine, citric acid, malic acid, gluconic acid, oxalic acid, succinic acid and other alkaline components with anti-mitotic effects. It enters the heart, liver, spleen, gallbladder and lung meridians. It can clear away heat and dampness, promote the flow of yang, stop bleeding, and has the effects of reducing weight and lowering blood lipids, anti-oxidation, anti-convulsion, protecting the liver, and resisting liver fibrosis.

[0014] Platycladus orientalis leaves: the dried branches and leaves of Platycladus orientalis, a plant of the Cupressaceae family. They are mostly harvested in summer and autumn and dried in the shade. They contain volatile oils such as anethole, camphor, bornyl acetate, terpene alcohols, and flavonoid components such as quercetin, ycigenin, kaempferol, and cypress biflavonoids. The leaves also contain trace elements such as potassium, sodium, nitrogen, phosphorus, calcium, magnesium, manganese, and zinc. Platycladus orientalis leaves enter the heart, liver, and large intestine meridians, and have the effects of stopping bleeding and cooling blood, relieving cough, removing phlegm, relieving asthma, and calming the nerves. They have inhibitory effects on Staphylococcus aureus, Catarrhalis, Shigella dysenteriae, Salmonella typhi, and Diphtheria.

[0015] Licorice polysaccharide: a type of α-D-pyranose extracted from licorice. Licorice, also known as beautiful grass, honey grass, Guolao, sweet grass, powder grass, stick grass, and Lingtong, is the root and rhizome of the leguminous plant licorice. Licorice has the effects of tonifying the spleen and replenishing qi, clearing away heat and detoxifying, relieving cough and reducing phlegm, harmonizing various medicines, and relieving pain. Licorice polysaccharide is one of the main components of the leguminous plant licorice. Studies have shown that licorice polysaccharide has a wide range of pharmacological effects and plays an important role in immune regulation, anti-oxidation, anti-tumor, antibacterial, anti-inflammatory and regulating intestinal flora.

[0016] Vitamin E: Also known as vitamin E, also known as tocopherol or tocopherol, it is one of the most important antioxidants. It has anti-aging, immunity enhancement and anti-tumor functions. Vitamin E has a wide range of effects in preventing and treating cardiovascular and cerebrovascular diseases, tumors, diabetes and other complications, central nervous system diseases, motor system diseases, skin diseases, etc.

[0017] Nano zinc oxide: It is a multifunctional new inorganic material with a particle size of 1 to 100 nanometers. Nano zinc oxide (nano-ZnO) has optical, electrical, magnetic, thermal, and sensitive properties that conventional zinc oxide does not have. The product is highly active and has anti-infrared, anti-ultraviolet and bactericidal functions. It has been widely used in sunscreen cosmetics, new functional fibers with antibacterial, deodorizing and anti-ultraviolet properties, self-cleaning antibacterial glass, ceramics, anti-infrared and anti-ultraviolet shielding materials, sanitary ware, sewage treatment and other products.

[0018] Preferably, it is composed of the following components in parts by weight: 15-20 parts of lotus leaves, 15-20 parts of Platycladus orientalis leaves, 10-15 parts of licorice polysaccharides, 0.3-0.5 parts of vitamin E, and 0.1-0.5 parts of nano zinc oxide or nano zinc oxide capped with paeonol.

[0019] The nano zinc oxide capped with the moutan bark extract (paeonol) means that in the process of forming zinc oxide nanoparticles, the active ingredients in the moutan bark extract, such as paeonol, capture zinc oxide molecules, and after the zinc oxide molecules form particles, they are capped on the particle surface to prevent the large-scale aggregation of nano zinc oxide particles. The present invention proves through experiments that the nano zinc oxide synthesized with the moutan bark extract as a template has better antibacterial function than the commercially available nano zinc oxide, and is green, pollution-free and environmentally friendly.

[0020] Further preferably, the synthesis process of the paeonol-capped nano zinc oxide is: paeonol and zinc acetate react in water under alkaline conditions to obtain paeonol-capped zinc oxide.

[0021] More preferably, the amount of zinc acetate used per 1 g of paeonol is 2 to 4 g.

[0022] More preferably, the average particle size of the nano zinc oxide capped with paeonol is 20 to 90 nm.

[0023] Specifically, the synthesis process is: 10 mL of 30% peony root bark extract (paeonol) solution is slowly added to 50 mL of 1 mol / L zinc acetate (ZnC4H6O4) solution, and reacted at pH = 12 to obtain paeonol-capped nano zinc oxide.

[0024] A method for preparing an anti-immune stress composition for chickens comprises the following steps: crushing lotus leaves and arborvitae leaves, and then mixing the crushed leaves with liquorice polysaccharide, vitamin E and nano zinc oxide or nano zinc oxide capped with paeonol.

[0025] The preparation method of the anti-immune stress composition for chickens of the present invention does not require the use of complex instruments and equipment, has simple steps, strong operability, and is easy for industrial large-scale production.

[0026] Application of an anti-immune stress composition for chickens in preparing feed for enhancing the anti-immune stress ability of chickens.

[0027] It has been verified that the anti-immune stress composition for chickens of the present invention can significantly improve the immune stress condition of chickens, improve body metabolism, effectively enhance the immunity of chickens, increase the feed intake of chickens under immune stress, stimulate the digestion and absorption function of the gastrointestinal tract of chickens, and enhance the ability of chickens to resist immune stress.

[0028] Preferably, the anti-immune stress composition for chickens is added to the feed of the complete feed formula at a mass percentage of 1-5%.

[0029] Beneficial effects of the present invention:

[0030] The invention adds nano zinc oxide or nano zinc oxide capped with paeonol to the anti-immune stress composition for chickens, which has a small particle size and high bioavailability. Compared with ordinary zinc oxide, the dosage is significantly reduced, which reduces the production cost. The reduction in dosage also reduces the zinc residue in the animal body and reduces the risk of metal poisoning. The addition amount is reduced and the absorption rate is high, which greatly reduces the discharge of zinc and reduces the pollution of heavy metals to the environment.

[0031] In the anti-immune stress composition for chickens of the present invention, the synergistic effect of nano zinc oxide or nano zinc oxide capped with paeonol and vitamin E promotes the metabolism of glucose, protein and lipid substances. At the same time, since nano zinc oxide or nano zinc oxide capped with paeonol has a small particle size and active properties, it is easy to generate free radicals, and vitamin E has the function of scavenging free radicals. The combination of vitamin E and nano zinc oxide achieves a balance between the generation and scavenging of free radicals, maintains the homeostasis of free radicals in the chicken body, reduces the risk of oxidative damage caused by the use of nano zinc oxide, and further improves the safety of the application of nano zinc oxide.

[0032] The anti-immune stress composition for chickens of the present invention adopts lotus leaf, arborvitae leaf, glycyrrhiza polysaccharide, vitamin E and nano zinc oxide or paeonol capped nano zinc oxide to play a synergistic effect. By targeting cyclooxygenase-2 (COX-2), COX-2 activity is inhibited, the generation of prostaglandin E2 is reduced, and the serum corticosterone level is reduced. By targeting nuclear factor E2-related factor 2 (Nrf2), the gene expression of its downstream factors reduced nicotinamide adenine dinucleotide phosphate: quinone oxidoreductase 1 (NQO-1), heme oxygenase-1 (HO-1), and SOD1 is increased, the activity of SOD and GSH-Px in chicken plasma, liver and small intestine can be increased, and the content of MDA in plasma, liver and small intestine can be reduced, thereby enhancing the antioxidant capacity of chickens. By reducing the levels of proinflammatory factors TNF-α, IL-1β and IL-6 in the plasma, liver and small intestine of immune stressed chickens, and increasing the levels of anti-inflammatory factors IL-2, IL-4, γ-IFN and immunoglobulins IgG, IgA and IgM, immune regulation of immune stressed chickens was performed. By increasing the height of villi and the villus-crypt ratio of the jejunum of immune stressed chickens, the depth of crypts and the thickness of the intestinal wall were reduced. The gene and protein expressions of intestinal barrier-related factors Claudin-1, Occludin and ZO-1 in the small intestine of chickens were increased, the intestinal barrier function was enhanced, the intestinal flora was regulated, the abundance of beneficial intestinal bacteria was increased, and the abundance of harmful intestinal bacteria was reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The green synthesis principle and process of nano zinc oxide capped with paeonol of the present invention;

[0034] Figure 2 This is a schematic diagram of nano zinc oxide capping the active ingredients such as paeonol of the present invention;

[0035] Figure 3 The figure is the SEM spectrum of the nano zinc oxide capped with paeonol of the present invention. DETAILED DESCRIPTION

[0036] The present invention is further described in detail below in conjunction with specific examples. Unless otherwise specified, the equipment and reagents used in each embodiment, experimental example and comparative example can be obtained from commercial sources.

[0037] The anti-immune stress composition for chickens of the present invention is prepared according to the following weight portions: 15-20 parts of lotus leaves, 15-20 parts of Platycladus orientalis leaves, 10-15 parts of licorice polysaccharide, 0.1-0.5 parts of vitamin E, and 0.05-0.5 parts of nano zinc oxide or nano zinc oxide capped with paeonol.

[0038] The preparation method of the anti-immune stress composition for chickens is as follows: after crushing lotus leaves and Platycladus orientalis leaves, the powder is mixed with licorice polysaccharide, vitamin E and nano zinc oxide or nano zinc oxide capped with paeonol.

[0039] The nano zinc oxide capped with paeonol in the present invention is synthesized with paeonol extract as a template. The synthesis principle is as follows: the active ingredient paeonol is present in the peony root bark extract. Paeonol contains phenolic hydroxyl groups and has reducing properties. It can be used as a bioreducing agent and capping agent in the biosynthesis of nanoparticles. 2+ Under the action of paeonol, it is reduced to zero-valent Zn, which combines with dissolved oxygen in water to form ZnO nanoparticles. Figure 1 During the formation of zinc oxide nanoparticles, the active ingredients in the moutan bark extract, such as paeonol, capture zinc oxide molecules and cap the surface of the particles after the zinc oxide molecules form particles, preventing the aggregation of nano zinc oxide particles. Figure 2 .

[0040] The synthesis process is as follows: 10 mL of 30% peony root bark extract (paeonol) solution is slowly dripped into 50 mL of 1 mol / L zinc acetate (ZnC4H6O4) solution, and NaOH is added to maintain the pH value at 12. The gel-like white precipitate is continuously stirred at room temperature for 2 hours. The white precipitate is washed with distilled water several times to remove impurities. The washed precipitate is dried in a microwave oven for 20 minutes, and paeonol-capped nano-zinc oxide is obtained after grinding. The average particle size of the paeonol-capped nano-zinc oxide synthesized greenly using paeonol as a template is 20 to 90 nm, and the SEM spectrum is shown in FIG. Figure 3 .

[0041] The method for using the chicken immune stress resistance composition of the present invention is to add the composition to the chicken feed of the complete feed formula in an amount of 1-5% by weight. The nano zinc oxide capped with paeonol in the chicken immune stress resistance composition is synthesized by the above-mentioned synthesis method.

[0042] 1. Specific examples of an anti-immune stress composition for chickens and a preparation method thereof

[0043] Example 1

[0044] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 15 parts of lotus leaves, 15 parts of Platycladus orientalis leaves, 15 parts of licorice polysaccharides, 0.5 parts of vitamin E, and 0.5 parts of nano zinc oxide capped with paeonol.

[0045] The preparation method of the anti-immune stress composition for chickens in this embodiment is as follows: weigh the components according to the above weight portions, grind the lotus leaves and Platycladus orientalis leaves by an ultrafine grinder, pass through a 200-mesh sieve, add the powders of glycyrrhizic polysaccharide, vitamin E, and paeonol-capped nano zinc oxide to the sieved powder, and mix well.

[0046] Example 2

[0047] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 15 parts of lotus leaves, 20 parts of Platycladus orientalis leaves, 10 parts of licorice polysaccharides, 0.5 parts of vitamin E, and 0.3 parts of nano zinc oxide capped with paeonol.

[0048] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0049] Example 3

[0050] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 20 parts of lotus leaves, 15 parts of Platycladus orientalis leaves, 15 parts of licorice polysaccharides, 0.3 parts of vitamin E, and 0.2 parts of nano zinc oxide capped with paeonol.

[0051] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0052] Example 4

[0053] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 15 parts of lotus leaves, 20 parts of Platycladus orientalis leaves, 15 parts of licorice polysaccharides, 0.3 parts of vitamin E, and 0.2 parts of nano zinc oxide capped with paeonol.

[0054] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0055] Example 5

[0056] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 20 parts of lotus leaves, 20 parts of Platycladus orientalis leaves, 15 parts of licorice polysaccharides, 0.3 parts of vitamin E, and 0.1 parts of nano zinc oxide capped with paeonol.

[0057] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0058] Example 6

[0059] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 15 parts of lotus leaves, 15 parts of Platycladus orientalis leaves, 10 parts of licorice polysaccharides, 0.1 parts of vitamin E, and 0.05 parts of nano zinc oxide capped with paeonol.

[0060] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0061] Example 7

[0062] The anti-immune stress composition for chickens of this embodiment is composed of the following components in parts by weight: 25 parts of lotus leaves, 25 parts of Platycladus orientalis leaves, 20 parts of licorice polysaccharides, 0.5 parts of vitamin E, and 0.5 parts of nano zinc oxide capped with paeonol.

[0063] The preparation method of the anti-immune stress composition for chickens in this example is the same as that described in Example 1.

[0064] 2. Related Experimental Examples Application of the Anti-Immune Stress Composition for Chickens in the Preparation of Feed for Enhancing the Anti-Immune Stress Ability of Chickens

[0065] Experimental Example 1: Verification of the antibacterial effect of nano zinc oxide capped with paeonol

[0066] In this experimental example, paeonol-capped nano zinc oxide was synthesized using moutan bark extract as a template, and its antibacterial effect compared with commercially available nano zinc oxide is shown in Table 1. As shown in Table 1, paeonol-capped nano zinc oxide is significantly better than commercially available nano zinc oxide in terms of antibacterial effect.

[0067] Table 1 Antibacterial effect of paeonol-capped nano zinc oxide and commercially available nano zinc oxide

[0068]

[0069]

[0070] Experimental Example 2: Verification of the synergistic effect of nano zinc oxide capped with vitamin E and paeonol

[0071] This experimental example uses AA broiler chickens, a total of 180 chickens, and 6 treatment groups are set, each group has 3 replicates, each replicate has 10 chickens, group I is the control group, group II is the nano zinc oxide group capped with paeonol (40 mg / kg), group III is the low vitamin E group (25 mg / kg), group IV is the high vitamin E group (50 mg / kg), group V is the nano zinc oxide capped with paeonol + low vitamin E group (40 mg / kg paeonol capped nano zinc oxide + 25 mg / kg vitamin E), group VI is the nano zinc oxide group capped with paeonol + high vitamin E group (40 mg / kg paeonol capped nano zinc oxide + 50 mg / kg vitamin E). Blood was collected from the wing vein on the 21st day, 3 ml / each, and the chicken serum antioxidant index was tested. The test results are shown in Table 2.

[0072] As shown in Table 2, the addition of paeonol-capped nano zinc oxide, vitamin E, and the mixed addition of paeonol-capped nano zinc oxide and vitamin E to the diet can significantly increase the activity of chicken CAT (p<0.01) and SOD (p<0.01), and improve the chicken's ability to inhibit hydroxyl free radicals (p<0.01) and resist superoxide anions (p<0.01), and the effect of adding paeonol-capped nano zinc oxide to the diet is better than that of adding vitamin E to the diet. The mixed addition of paeonol-capped nano zinc oxide and vitamin E can reduce the levels of H2O2 (p<0.01) and MDA (p<0.01), and improve the activity of CAT (p<0.01) and SOD (p<0.01), and the ability to inhibit hydroxyl free radicals (p<0.01) and resist superoxide anions (p<0.01) is better than the addition of nano zinc oxide and vitamin E alone.

[0073] Table 2 Free radical scavenging and antioxidant effects of nano zinc oxide capped with vitamin E and paeonol

[0074]

[0075] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), different uppercase letters indicate extremely significant differences (P<0.01), and the same letters indicate insignificant differences (P>0.05).

[0076] To further confirm that vitamin E and nano zinc oxide have a synergistic effect, 60 AA broilers were used, and 4 treatment groups were set up, with 15 in each group. Group I was the control group, Group II was the nano zinc oxide group capped with paeonol (40 mg / kg), Group III was the vitamin E group (40 mg / kg), and Group IV was the nano zinc oxide + vitamin E group capped with paeonol (30 mg / kg paeonol-capped nano zinc oxide + 10 mg / kg vitamin E). On the 14th day, 2 ml of blood was collected from the wing vein of each chicken to test the antioxidant index of chicken serum. The test results are shown in Table 3.

[0077] As shown in Table 3, the addition of paeonol-capped nano zinc oxide, vitamin E, and the mixed addition of paeonol-capped nano zinc oxide and vitamin E to the diet can significantly increase the activity of chicken plasma CAT and SOD, improve the chicken's ability to inhibit hydroxyl free radicals and resist superoxide anions, and reduce the levels of H2O2 and MDA. The addition of 30 mg / kg nano zinc oxide + 10 mg / kg vitamin E to the diet has a similar antioxidant effect as the addition of 40 mg / kg paeonol-capped nano zinc oxide to the diet. This shows that vitamin E and paeonol-capped nano zinc oxide have a synergistic effect of scavenging free radicals and anti-oxidation.

[0078] Table 3 Synergistic free radical scavenging and antioxidant effects of nano zinc oxide capped with vitamin E and paeonol

[0079] Group I Group II Group III Group IV CAT (U / mL) <![CDATA[5.23±0.91 bB ]]> <![CDATA[12.83±2.17 A ]]> <![CDATA[7.61±1.36 aB ]]> <![CDATA[15.05±3.82 A ]]> SOD(U / mL) <![CDATA[9.29±1.18 bB ]]> <![CDATA[18.63±3.27 A ]]> <![CDATA[11.62±2.07 aB ]]> <![CDATA[20.15±4.38 A ]]> <![CDATA[H2O2(mmol / L)]]> <![CDATA[63.28±10.16 a ]]> <![CDATA[50.27±13.14 b ]]> <![CDATA[60.55±19.32 a ]]> <![CDATA[47.39±9.25 b ]]> Hydroxyl free radical inhibition ability (U / mL) <![CDATA[30.67±4.28 b ]]> <![CDATA[43.64±5.29 a ]]> <![CDATA[33.82±5.07 b ]]> <![CDATA[45.26±7.03 a <!-- 6 -->]]> Anti-superoxide anion ability (U / L) <![CDATA[102.36±18.15 b ]]> <![CDATA[119.37±13.86 a ]]> <![CDATA[107.49±16.97 ab ]]> <![CDATA[116.79±13.42 a ]]> MDA (nmol / mL) <![CDATA[6.28±0.37 a ]]> <![CDATA[4.89±0.57 b ]]> <![CDATA[5.82±1.07 ab ]]> <![CDATA[4.91±1.20 b ]]>

[0080] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), different uppercase letters indicate extremely significant differences (P<0.01), and the same letters indicate insignificant differences (P>0.05).

[0081] Experimental Example 3

[0082] In this experimental example, AA broiler chickens were used as research subjects, lipopolysaccharide (LPS) was injected to establish an immune stress model, and the application effect of the anti-immune stress composition for chickens in Example 3 was verified. The specific operation was as follows:

[0083] A total of 240 AA broilers were randomly divided into 4 treatments, with 6 replicates in each treatment and 10 chickens in each replicate. Treatment I was a saline group (fed with a normal diet), treatment II was an LPS group (fed with a normal diet), treatment III was an LPS+anti-immune stress composition group (fed with a diet supplemented with 1% anti-immune stress composition), and treatment IV was an LPS+paeonol-capped nano-zinc oxide group (fed with a diet supplemented with 40 mg / kg paeonol-capped nano-zinc oxide, which was consistent with the amount of paeonol-capped nano-zinc oxide in group III). Starting from 14 days of age, the LPS group, LPS+anti-immune stress composition group, and LPS+paeonol-capped nano-zinc oxide group were intraperitoneally injected with a dose of 0.5 mg / kg of LPS, and the saline group was injected with an equal amount of saline. Weigh and record the food intake before injection and on the 1st, 3rd, 5th, 7th and 9th day after injection. Select one from each replicate, collect 3ml of blood from the wing vein, and collect tissues and organs such as the hypothalamus and liver. The test indicators are as follows:

[0084] Serum immunoglobulin IgA, IgM, IgG levels, cytokine γ-IFN, IL-2, IL-4, IL-1, IL-6, TNF-α levels, serum COX-2, prostaglandin E2, corticosterone levels. SOD, GSH-Px activities, MDA content in serum, liver and small intestine. Gene expression of Nrf2 and its downstream factors NQO-1, HO-1, SOD1, and gene expression of small intestinal barrier-related factors Claudin-1, Occludin and ZO-1. Metagenomics detection of intestinal microbial diversity and abundance.

[0085] The test results are as follows:

[0086] As shown in Table 4, immune stress significantly reduces the average daily weight gain and average daily feed intake of broilers, and significantly improves the feed-to-weight ratio. Adding the anti-immune stress composition or paeonol-capped nano zinc oxide to the diet significantly improves the average daily weight gain of broilers with immune stress and reduces the feed-to-weight ratio. In addition, the application effect of the anti-immune stress composition of the present invention is better than that of nano zinc oxide capped with paeonol alone.

[0087] Table 4 Average daily weight gain, average daily feed intake, and feed-to-weight ratio of broilers in each treatment group

[0088] Average daily weight gain (g / d) Average daily feed intake (g / d) Material weight ratio Treatment I <![CDATA[68.38±1.26 a ]]> <![CDATA[119.72±2.85 a ]]> 1.75 Process II <![CDATA[48.53±2.35 c ]]> <![CDATA[106.25±5.93 b ]]> 2.18 Process III <![CDATA[60.29±3.14 b ]]> <![CDATA[115.51±4.83 a ]]> 1.91 Treatment IV <![CDATA[58.38±1.26 b ]]> <![CDATA[110.33±3.95 a ]]> 1.89

[0089] Note: Different lowercase letters in the upper right corner of the same column indicate significant differences (P<0.05), while the same letters indicate insignificant differences (P>0.05).

[0090] As shown in Table 5, adding the anti-immune stress composition or paeonol-capped nano zinc oxide to the diet significantly improved the IgA, IgM, and IgG levels of broilers with immune stress, among which the application effect of the anti-immune stress composition of the present invention was better than that of paeonol-capped nano zinc oxide alone.

[0091] Table 5 Serum immunoglobulin levels of 42-day-old chickens in each treatment group (mg / ml)

[0092] IgA IgM IgG Treatment I <![CDATA[4.74±0.81 b ]]> <![CDATA[24.94±2.32 b ]]> <![CDATA[26.35±2.02 b ]]> Process II <![CDATA[4.19±1.00 b ]]> <![CDATA[27.84±3.00 b ]]> <![CDATA[17.77±5.02 c <!-- 7 -->]]> Process III <![CDATA[6.39±1.16 a ]]> <![CDATA[32.95±2.48 a ]]> <![CDATA[58.78±5.14 a ]]> Treatment IV <![CDATA[5.28±1.09 a ]]> <![CDATA[28.95±3.65 b ]]> <![CDATA[49.61±5.23 a ]]>

[0093] Note: Different lowercase letters in the upper right corner of the same column indicate significant differences (P<0.05), while the same letters indicate insignificant differences (P>0.05).

[0094] As shown in Table 6, adding the anti-immune stress composition or the nano zinc oxide capped with paeonol to the diet significantly increased the levels of anti-inflammatory factors γ-IFN, IL-2, and IL-4 in broilers with immune stress, and reduced the levels of pro-inflammatory factors IL-1β, IL-6, and TNF-α. Among them, the application effect of the anti-immune stress composition of the present invention is better than that of the nano zinc oxide capped with paeonol alone.

[0095] Table 6 Serum cytokine content of 42-day-old chickens (pg / ml)

[0096] γ-IFN IL-2 IL-4 IL-1β IL-6 TNF-α Treatment I <![CDATA[298.67±13.51 b ]]> <![CDATA[24.19±5.96 a ]]> <![CDATA[102.81±10.59 c ]]> <![CDATA[322±10.11 b ]]> <![CDATA[96.62±4.56 b ]]> <![CDATA[411±13.25 b ]]> Process II <![CDATA[294.79±9.58 b ]]> <![CDATA[20.20±4.22 a ]]> <![CDATA[97.16±9.69 c ]]> <![CDATA[401±8.57 a ]]> <![CDATA[128.23±8.05 a ]]> <![CDATA[532±20.67 a ]]> Process III <![CDATA[353.77±15.31 a ]]> <![CDATA[25.72±3.61 a ]]> <![CDATA[127.89±7.92 a ]]> <![CDATA[305±9.09 c ]]> <![CDATA[95.12±3.94 b ]]> <![CDATA[409±15.85 b ]]> Treatment IV <![CDATA[331.58±10.03 a ]]> <![CDATA[23.22±4.01 a ]]> <![CDATA[117.09±7.02 b ]]> <![CDATA[336±7.62 b ]]> <![CDATA[100.12±7.33 b ]]> <![CDATA[421±10.51 b ]]>

[0097] Note: Different lowercase letters in the upper right corner of the same column indicate significant differences (P<0.05), while the same letters indicate insignificant differences (P>0.05).

[0098] As shown in Table 7, adding the anti-immune stress composition or paeonol-capped nano zinc oxide to the diet significantly increased the SOD activity and reduced the MDA content of broilers with immune stress. The application effect of the anti-immune stress composition of the present invention is better than that of paeonol-capped nano zinc oxide alone.

[0099] Table 7 Serum biochemical parameters of 42-day-old chickens in each treatment group

[0100]

[0101] Note: Different lowercase letters in the upper right corner of the same column indicate significant differences (P<0.05), while the same letters indicate insignificant differences (P>0.05).

[0102] As shown in Table 8, adding the anti-immune stress composition or the nano zinc oxide capped with paeonol to the diet significantly reduced the COX2, prostaglandin E2, and corticosterone content of broilers with immune stress, and relieved immune stress. Among them, the application effect of the anti-immune stress composition of the present invention is better than that of the nano zinc oxide capped with paeonol alone.

[0103] Table 8 Serum COX2, prostaglandin E2, and corticosterone levels (ng / mL) of 42-day-old chickens in each treatment group

[0104] Treatment I Process II Process III Treatment IV COX-2 <![CDATA[12.82±0.92 c ]]> <![CDATA[35.17±3.67 a ]]> <![CDATA[13.93±1.95 c ]]> <![CDATA[20.00±3.11 b ]]> Prostaglandin E2 <![CDATA[35.76±5.32 b ]]> <![CDATA[55.72±8.61 a ]]> <![CDATA[37.89±7.92 b ]]> <![CDATA[40.00±3.09 b ]]> Corticosterone <![CDATA[1.44±0.21 b ]]> <![CDATA[2.19±0.09 a ]]> <![CDATA[1.39±0.16 b ]]> <![CDATA[1.58±0.09 b ]]>

[0105] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), while the same letters indicate no significant differences (P>0.05).

[0106] As shown in Table 9, adding the anti-immune stress composition or paeonol-capped nano zinc oxide to the diet significantly increased the expression levels of Nrf2 and its downstream factors HO-1 and SOD1 mRNA in immune-stressed broilers, and improved the antioxidant capacity. Among them, the application effect of the anti-immune stress composition of the present invention is better than that of paeonol-capped nano zinc oxide alone.

[0107] Table 9 Relative expression of Nrf2, HO-1 and SOD1 genes in the liver of 42-day-old chickens in each treatment group

[0108]

[0109]

[0110] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), while the same letters indicate no significant differences (P>0.05).

[0111] As shown in Table 10, adding the anti-immune stress composition or the nano zinc oxide capped with paeonol to the diet significantly increased the gene expression of the intestinal barrier factors Occludin, Claudin-1, and ZO-1 in chickens with immune stress, and maintained the intestinal barrier function. Among them, the application effect of the anti-immune stress composition of the present invention is better than that of the nano zinc oxide capped with paeonol alone.

[0112] Table 10 Gene expression of small intestinal barrier factors Occludin, Claudin-1 and ZO-1 in 42-day-old chickens in each treatment group

[0113] Treatment I Process II Process III Treatment IV Occludin <![CDATA[1.08±0.03 b ]]> <![CDATA[1.07±0.06 b ]]> <![CDATA[1.85±0.62 a ]]> <![CDATA[1.46±0.18 a ]]> Claudin-1 <![CDATA[1.00±0.02 ab ]]> <![CDATA[1.02±0.21 ab ]]> <![CDATA[1.50±0.32 a ]]> <![CDATA[1.36±0.31 a ]]> ZO-1 <![CDATA[1.02±0.05 b ]]> <![CDATA[1.09±0.19 b ]]> <![CDATA[1.89±0.26 a ]]> <![CDATA[1.47±0.22 ab ]]>

[0114] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), while the same letters indicate no significant differences (P>0.05).

[0115] As shown in Table 11, adding the anti-immune stress composition or paeonol-capped nano zinc oxide to the diet significantly increased the α diversity of the intestinal flora of immune-stressed broilers. The application effect of the anti-immune stress composition of the present invention is better than that of paeonol-capped nano zinc oxide alone.

[0116] Table 11 Diversity of intestinal bacteria

[0117] Shannon index ACE index Chao1 index Treatment I <![CDATA[6.41±0.63 a ]]> <![CDATA[478.48±26.25 ab ]]> <![CDATA[479.80±33.27 ab ]]> Process II <![CDATA[5.98±0.59 a ]]> <![CDATA[399.39±22.83 b ]]> <![CDATA[336.56±25.62 b ]]> Process III <![CDATA[6.90±0.49 a ]]> <![CDATA[569.29±17.39 a ]]> <![CDATA[557.25±26.19 a ]]> Treatment IV <![CDATA[6.86±0.65 a ]]> <![CDATA[524.01±38.51 ab ]]> <![CDATA[509.82±35.45 ab ]]>

[0118] Note: Different lowercase letters in the upper right corner of the same column indicate significant differences (P<0.05), while the same letters indicate insignificant differences (P>0.05).

[0119] The dominant flora in the intestine of broilers mainly include Firmicutes and Bacteroidetes. As shown in Table 12, adding the anti-immune stress composition or nano zinc oxide capped with paeonol to the diet significantly increases the levels of Firmicutes and Bacteroidetes in the intestine of broilers with immune stress, and reduces the levels of Proteobacteria and Actinobacteria. Among them, the application effect of the anti-immune stress composition of the present invention is better than that of nano zinc oxide capped with paeonol alone.

[0120] Table 12 Relative abundance of intestinal bacteria at the phylum level in broiler chickens in each treatment group

[0121]

[0122]

[0123] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), while the same letters indicate no significant differences (P>0.05).

[0124] As shown in Table 13, adding the anti-immune stress composition or the nano zinc oxide capped with paeonol to the diet significantly increased the level of beneficial bacteria Lactobacillus in the intestine of broiler chickens and reduced the level of harmful bacteria Escherichia. The application effect of the anti-immune stress composition of the present invention is better than that of the nano zinc oxide capped with paeonol alone.

[0125] Table 13 Relative abundance of intestinal bacteria at the genus level

[0126] Treatment I Process II Process III Treatment IV Lactobacillus <![CDATA[12.50±4.62 b ]]> <![CDATA[5.45±1.83 c ]]> <![CDATA[16.18±4.59 a ]]> <![CDATA[15.50±3.83 a ]]> Unclassified Lachnospiraceae related genera <![CDATA[6.23±1.63 ab ]]> <![CDATA[5.21±1.38 b ]]> <![CDATA[8.14±1.72 a ]]> <![CDATA[8.07±1.36 a ]]> Escherichia <![CDATA[0.76±0.08 b ]]> <![CDATA[2.63±1.03 a ]]> <![CDATA[0.53±0.12 b ]]> <![CDATA[0.62±0.29 b ]]>

[0127] Note: Different lowercase letters in the upper right corner of the same line indicate significant differences (P<0.05), while the same letters indicate no significant differences (P>0.05).

[0128] Finally, it is noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An anti-immune stress composition for chickens, characterized in that: The invention comprises the following components in parts by weight: 15-25 parts of lotus leaves, 15-25 parts of Platycladus orientalis leaves, 10-20 parts of liquorice polysaccharide, 0.1-0.5 parts of vitamin E and 0.05-0.5 parts of paeonol-capped nano zinc oxide; the synthesis process of the paeonol-capped nano zinc oxide is as follows: paeonol and zinc acetate react in water under alkaline conditions to obtain the paeonol-capped nano zinc oxide; the dosage of zinc acetate corresponding to 1 g of paeonol is 2-4 g; and the average particle size of the paeonol-capped nano zinc oxide is 20-90 nm.

2. The anti-immune stress composition for chicken according to claim 1, characterized in that: The invention is composed of the following components in parts by weight: 15-20 parts of lotus leaves, 15-20 parts of Platycladus orientalis leaves, 10-15 parts of licorice polysaccharide, 0.3-0.5 parts of vitamin E and 0.1-0.5 parts of nano zinc oxide capped with paeonol.

3. A method for preparing the anti-immune stress composition for chickens as claimed in claim 1 or 2, characterized in that: Lotus leaves and Platycladus orientalis leaves are crushed and mixed with licorice polysaccharide, vitamin E and nano zinc oxide capped with paeonol.

4. Use of the anti-immune stress composition for chickens as claimed in claim 1 or 2 in preparing feed for enhancing the anti-immune stress ability of chickens.

5. Use of the anti-immune stress composition for chickens according to claim 4 in preparing feed for enhancing the anti-immune stress ability of chickens, characterized in that: The anti-immune stress composition for chickens is added into the feed of the complete feed formula at a mass percentage of 1-5%.

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