An immune enhancer for laying hens and a preparation method thereof

Abstract

The present application belongs to the technical field of medicine or feed, and particularly relates to an immune enhancer for laying hens and a preparation method thereof. The immune enhancer for laying hens comprises: kirengeshomine, wolfberry polysaccharide, guar gum, vitamin E, vitamin B12, L-arginine, methionine and lysine. The immune enhancer for laying hens significantly improves the immune indexes and production performance of laying hens, provides safe and efficient immune protection for the laying hens, simultaneously reduces the breeding cost, is suitable for the intensive production mode, and has significant economic and social benefits.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical or feed technology, specifically relating to an immune enhancer for laying hens and its preparation method. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Egg-laying hen farming is a crucial component of agricultural production, with the health and productivity of hens directly impacting egg quality and economic benefits. Under intensive farming models, laying hens often face multiple pressures, including high-density rearing, environmental stress, and pathogen invasion, leading to weakened immunity and increased susceptibility to infectious diseases such as Newcastle disease and avian influenza. This results in decreased egg production, increased mortality, and even public health problems. Therefore, how to safely and effectively enhance the immunity of laying hens has become a pressing technical challenge for the poultry industry.

[0004] Traditional solutions rely heavily on antibiotics and chemically synthesized drugs, but face three major technological bottlenecks: (1) Drug resistance problem: continuous use increases the rate of drug resistance gene carriers.

[0005] (2) Residue risk: Antibiotics may remain in eggs.

[0006] (3) Limited effect: Although single natural active ingredients, such as polysaccharides, have certain immunomodulatory effects, their effects are limited when used alone.

[0007] Therefore, developing natural, safe, and effective immune enhancers has become an important direction for the current aquaculture industry.

[0008] Chinese patent CN109258980A discloses an organic feed and its formulation method that can improve the immunity of laying hens. The feed includes: corn, soybean meal, cottonseed meal, dicalcium phosphate, fish meal, limestone powder, calcium propionate, potato tubers, sweet potato roots, wheat bran, garlic, houttuynia cordata, selenium, a vitamin composition, a trace element composition, traditional Chinese medicine additives, a sugar composition, and an amino acid composition. This feed can improve the immune function of laying hens and ensure normal egg production.

[0009] Chinese patent CN107242383A discloses a feed for broilers that promotes digestion, appetite, and growth. Its ingredients include: corn flour, wheat bran, rapeseed meal, cockroach powder, krill powder, seaweed powder, xylooligosaccharides, wolfberry polysaccharides, maifanite powder, zeolite powder, acetic acid, fumaric acid, amino acid chelates, taurine, barley leaf powder, compound vitamins, compound protease preparations, vegetable oil, and traditional Chinese medicine additives. This feed can improve the gastrointestinal function of broilers, enhance their immunity, reduce morbidity, strengthen their digestive and absorptive capacity, improve feed conversion rate, and promote their growth and development.

[0010] However, the existing technologies mentioned above have complex production processes and high costs, and the mechanisms of action among the core active ingredients are not clearly defined, resulting in limited effectiveness in improving the immunity of laying hens. Summary of the Invention

[0011] To overcome the shortcomings of existing technologies, this invention provides an immune enhancer for laying hens and its preparation method. Its core components are a synergistic combination of natural active ingredients and nutritional supplements, including: crystalline glycosides, wolfberry polysaccharides, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine. The proportions by weight are: crystalline glycosides 0.1-0.5 parts, wolfberry polysaccharides 3.0-8.0 parts, guar gum 0.3-0.6 parts, vitamin E 0.1-0.4 parts, vitamin B12 0.1-0.5 parts, L-arginine 0.2-0.5 parts, methionine 0.2-0.5 parts, and lysine 0.2-0.5 parts.

[0012] Among them, crystallized blue glycoside (purity ≥98%, purchased from Shanghai Yuanye Biotechnology Co., Ltd.), wolfberry polysaccharide (purchased from Shaanxi Tianjingrui Biotechnology Co., Ltd.), and yucca saponin (purchased from Xi'an Weiao Biotechnology Co., Ltd.) were used.

[0013] This invention allows the above-mentioned active ingredients to be mixed and added to feed, which can improve the immunity of laying hens and increase the feed conversion ratio. Furthermore, the effectiveness of this laying hen immune enhancer can be further improved through a process using cyclodextrin encapsulation technology. This process involves encapsulating the active ingredients with hydroxypropyl-β-cyclodextrin to enhance the immune-enhancing effect in laying hens.

[0014] Specifically, the preparation method is as follows: (1) Dissolve cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in ethanol to obtain vitamin E ethanol solution; dissolve guar gum in water and adjust the pH to 5.0~6.0 to obtain guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly and sonicate. (4) After sonication, let the solution stand for 1-3 hours or refrigerate overnight, centrifuge, wash with cold water, and dry to obtain the final product.

[0015] Furthermore, the method of using the egg-laying hen immune enhancer is to add it to the feed.

[0016] In addition, the present invention also provides the application of the above-mentioned immune enhancer for laying hens in improving the quality of egg production.

[0017] Compared with the prior art, the technical advantages of the present invention are as follows: (1) In this invention, crystallized glycoside is applied to the field of immune enhancement in laying hens. Experiments have shown that it can specifically enhance mucosal immunity (IgA) and early immune response (IgM), complementing wolfberry polysaccharide and enhancing humoral immunity (IgG).

[0018] (2) Synergistic effect of guar gum and cyclodextrin: Guar gum optimizes the encapsulation efficiency of cyclodextrin by adjusting the pH of the solution, which significantly improves the utilization of active ingredients and enhances efficacy.

[0019] (3) This invention significantly improves the immune indicators and production performance of laying hens, provides safe and efficient immune protection for laying hens, and reduces breeding costs. It is suitable for intensive production mode and has significant economic and social benefits. Attached Figure Description

[0020] Figure 1 : The content of immunoglobulin IgG in the serum of laying hens.

[0021] Figure 2 : IgA content in the serum of laying hens.

[0022] Figure 3 : IgM immunoglobulin content in laying hen serum. Detailed Implementation

[0023] To make the objectives and technical solutions of this invention clearer, the following embodiments are provided for further explanation. However, the scope of protection of this invention is not limited to these embodiments; the embodiments are merely for illustrative purposes. Those skilled in the art should understand that any changes or equivalent substitutions that do not depart from the concept of this invention are included within the scope of protection of this invention.

[0024] Example 1: Immune enhancer for laying hens formula: Preparation process: The crystal blue glycoside, wolfberry polysaccharide, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0025] Example 2: Immune enhancer for laying hens formula: Preparation process: The crystal blue glycoside, wolfberry polysaccharide, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0026] Example 3: Immune Enhancer for Laying Hens formula: Preparation process: The crystal blue glycoside, wolfberry polysaccharide, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0027] Example 4: Immune Enhancer for Laying Hens formula: Preparation process: (1) Dissolve hydroxypropyl-β-cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in 10 ml of ethanol to obtain vitamin E ethanol solution; dissolve guar gum in 10 ml of water and adjust the pH to 5.5 with citric acid to obtain guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly, and sonicate (time is 25 min, temperature is 10℃). (4) After sonication, the solution is allowed to stand for 2 hours, centrifuged, washed with cold water, and vacuum dried at 50°C until the moisture content is <1%.

[0028] Example 5: Immune Enhancer for Laying Hens formula: Preparation process: (1) Dissolve hydroxypropyl-β-cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in 10 ml of ethanol to obtain vitamin E ethanol solution; dissolve guar gum in 10 ml of water and adjust the pH to 5.0 with sodium citrate to obtain guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly, and sonicate (time is 20 min, temperature is 5℃). (4) After sonication, the solution is allowed to stand for 3 hours, centrifuged, washed with cold water, and vacuum dried at 40°C until the moisture content is <1%.

[0029] Example 6: Immune Enhancer for Laying Hens formula: Preparation process: (1) Dissolve hydroxypropyl-β-cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in 10 ml of ethanol to obtain vitamin E ethanol solution; dissolve guar gum in 10 ml of water and adjust the pH to 6.0 with citric acid to obtain guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly, and sonicate (time is 30 min, temperature is 15℃). (4) After sonication, the solution is refrigerated at 2°C overnight, centrifuged, washed with cold water, and vacuum dried at 60°C until the moisture content is <1%.

[0030] Comparative Example 1: Immune Enhancer for Laying Hens formula: Preparation process: The ingredients of wolfberry polysaccharide, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0031] Comparative Example 2: Immune Enhancer for Laying Hens formula: Preparation process: The yucca saponins, wolfberry polysaccharides, guar gum, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0032] Comparative Example 3: Immune Enhancer for Laying Hens formula: Preparation process: The crystal blue glycoside, wolfberry polysaccharide, vitamin E, vitamin B12, L-arginine, methionine, and lysine are passed through an 80-mesh sieve and mixed evenly to obtain the final product.

[0033] Comparative Example 4: Immune Enhancer for Laying Hens formula: Preparation process: (1) Dissolve hydroxypropyl-β-cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in 10 ml of ethanol to obtain a vitamin E ethanol solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution, stir evenly, and sonicate (time is 25 min, temperature is 10℃). (4) After sonication, the solution is allowed to stand for 2 hours, centrifuged, washed with cold water, and vacuum dried at 50°C until the moisture content is <1%.

[0034] Comparative Example 5: Immune Enhancer for Laying Hens formula: Preparation process: (1) Dissolve hydroxypropyl-β-cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in 10 ml of ethanol to obtain a vitamin E ethanol solution; dissolve guar gum in 10 ml of water (pH about 7.0) to obtain a guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly, and sonicate (time is 25 min, temperature is 10℃). (4) After sonication, the solution is allowed to stand for 2 hours, centrifuged, washed with cold water, and vacuum dried at 50°C until the moisture content is <1%.

[0035] Pharmacodynamic experiments of the immune enhancer for laying hens of this invention laboratory animals Breed: Brown-shelled Hy-Line laying hens (peak egg production, weight 1.8-2.0 kg, age 180±5 days).

[0036] Number: 30 animals per group, 13 groups in total (namely blank control group, Example 1 group, Example 2 group, Example 3 group, Example 4 group, Example 5 group, Example 6 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group, Comparative Example 5 group, and EggVelodeon group), totaling 390 animals.

[0037] Feeding conditions: Standardized chicken house (temperature 22±2℃, humidity 50%~60%, light 16h / day), free access to feed and water, and standardized basic diet formula.

[0038] Dosing regimen Dosage: For Example 1 to Example 6 groups and Comparative Examples 1 to Comparative Examples 5 groups, 0.1% immune enhancer was added to the feed. For the EggVeile group, the dosage was added according to the instructions.

[0039] Method: Mix evenly into the daily diet and feed continuously for 6 weeks.

[0040] detection indicators Daily record indicators: daily feed intake, fecal condition.

[0041] Serum immunoglobulins: IgG, IgA, and IgM levels were detected by ELISA.

[0042] Production performance indicator: feed conversion ratio.

[0043] Pharmacodynamic experimental results Daily feed intake of laying hens Table 1 Daily feed intake and fecal condition of laying hens Table 1 shows the average daily feed intake and fecal condition of laying hens in Examples 1-6, Comparative Examples 1-5, and the EggVitamin group. Compared with the blank control group, the feed intake of laying hens in each example group and comparative example group was within the normal range, while the feed intake of laying hens in the EggVitamin group was slightly higher. The fecal condition of laying hens in all groups was normal, and the intestinal microbiota of laying hens was stable. This indicates that the immune enhancer for laying hens of the present invention has no negative impact on the intestinal health of laying hens, providing a safety basis for its clinical application.

[0044] Serum immunoglobulin IgG, IgA, and IgM levels Table 2 Serum Immunoglobulin IgG, IgA, and IgM levels Table 1 and Figure 1 , Figure 2 , Figure 3The levels of immunoglobulins IgG, IgA, and IgM in the serum of laying hens were measured. IgG, the most abundant immunoglobulin in serum (approximately 75-80%), is the mainstay of humoral immunity. It neutralizes pathogen toxins, activates the complement system, and clears pathogens through antibody-dependent cell-mediated cytotoxicity (ADCC). Compared to the blank model group, the IgG levels in laying hens in Examples 1-6 were significantly increased (P < 0.01). IgA is mainly found on mucosal surfaces (such as the respiratory and digestive tracts) and is the first line of defense in mucosal immunity, acting by preventing pathogen adhesion and neutralizing viruses. The IgA levels in laying hens in Examples 1-6 were significantly different from those in the blank model group (P < 0.01). IgM is the earliest antibody produced in the primary immune response. It has a large molecular weight, is mainly distributed in the blood, and efficiently activates the complement system. Compared to the blank model group, the IgM levels in the serum of laying hens in Examples 1-6 were significantly different (P < 0.01). Analysis showed that hydroxypropyl-β-cyclodextrin encapsulation, in synergistic effect with guar gum, significantly increased immunoglobulin levels and enhanced humoral immune responses. The IgA / IgM levels in Comparative Examples 1 and 2, compared to Example 1, were decreased, indicating that crystallizing glycosides have a specific regulatory effect on mucosal immunity and early immune responses.

[0045] Egg ratio The feed conversion ratio (FCR) of laying hens is an important indicator for measuring the relationship between feed consumption and egg production per unit of time, reflecting the amount of feed consumed by a hen to produce one egg.

[0046] Total feed consumption (kg) refers to the total amount of feed consumed by a flock of laying hens within a certain period of time. Total egg production (kg) refers to the total weight of eggs produced by a flock of laying hens within the same period of time. Feed conversion ratio = Total feed consumption / Total egg weight.

[0047] Table 3 Feed conversion ratio for laying hens Table 3 shows that the feed conversion ratio of the laying hens in Examples 1 to 6 of this invention is significantly lower than that of the blank control group, the commercially available EggVeile group, and each comparative group. Taking Example 6 as an example, the feed conversion ratio of 1.85 has reached the industry-leading level and is suitable for large-scale farms to reduce overall costs. For example, based on 10,000 chickens, each chicken saves about 4.8g of feed per day, and saves about 23,000 yuan in costs over the whole cycle.

[0048] All data were analyzed using SPSS 22.0 software. Quantitative data were expressed as follows: The differences among multiple groups were analyzed using one-way ANOVA, and the comparisons between two groups were performed using independent samples t-tests. The significance level was set at P < 0.05.

Claims

1. An immune enhancer for laying hens, characterized in that, The egg-laying hen immune enhancer, calculated by weight, comprises: 0.1-0.5 parts of crystallizing glycoside, 3.0-8.0 parts of wolfberry polysaccharide, 0.3-0.6 parts of guar gum, 0.1-0.4 parts of vitamin E, 0.1-0.5 parts of vitamin B12, 0.2-0.5 parts of L-arginine, 0.2-0.5 parts of methionine, 0.2-0.5 parts of lysine, and 1-2 parts by weight of cyclodextrin of the above ingredients; The preparation method of the egg-laying hen immune enhancer is as follows: (1) Dissolve cyclodextrin in water to prepare a saturated solution; (2) Dissolve vitamin E in ethanol to obtain vitamin E ethanol solution; dissolve guar gum in water and adjust the pH to 5.0~6.0 with citric acid or citrate to obtain guar gum solution; (3) Add crystallizing glycoside, wolfberry polysaccharide, vitamin B12, L-arginine, methionine and lysine to the saturated solution in step (1), then add vitamin E ethanol solution and guar gum solution, stir evenly and sonicate. (4) After sonication, let the solution stand for 1-3 hours or refrigerate overnight, centrifuge, wash with cold water, and dry to obtain the final product.

2. The egg-laying hen immune enhancer according to claim 1, characterized in that, The immune enhancer for laying hens, by weight, consists of: 0.3 parts crystallizing glycoside, 5.0 parts wolfberry polysaccharide, 0.4 parts guar gum, 0.3 parts vitamin E, 0.3 parts vitamin B12, 0.3 parts L-arginine, 0.3 parts methionine, and 0.3 parts lysine.

3. The egg-laying hen immune enhancer according to claim 1, characterized in that, The cyclodextrin is hydroxypropyl-β-cyclodextrin.

4. The egg-laying hen immune enhancer according to claim 1, characterized in that, The ultrasound duration is 20-30 minutes, and the temperature is 5℃-15℃.

5. The egg-laying hen immune enhancer according to claim 1, characterized in that, The drying process is vacuum drying at 40℃~60℃.

6. The egg-laying hen immune enhancer according to claim 1, characterized in that, The method of using the egg-laying hen immune enhancer is to add it to the feed.

7. The application of the egg-laying hen immune enhancer of claim 1 in improving the feed conversion ratio of laying hens.

Images