Compound traditional Chinese medicine composition for improving immunity of poultry and preparation method of composition

Abstract

The invention relates to the technical field of traditional Chinese medicine composition processing, in particular to a compound traditional Chinese medicine composition for improving immunity of poultry and a preparation method of the composition. Aiming at the physiological characteristics of short digestive tract and short food absorption time of the poultry, a basic formula with qi tonifying as a main part and digestion promoting as an assistant part is developed; a new composition is developed by optimizing a self-created basic prescription through technical means such as flavoring, odor reduction and substitution, so that the immune function is remarkably improved, the double effects of effectively reducing the standard deviation of chicken serum IgG and reducing the dispersion degree of data in a group are achieved, the overall improvement of group immunity is achieved, the immune effect of vaccines can be enhanced, and the application prospect is wide. Meanwhile, the dosage of the improved prescription is reduced by 33.3% compared with that of the basic prescription, the medication cost is reduced, in addition, by optimizing the polysaccharide water extraction process, the polysaccharide alcohol extraction process is effectively replaced, the problems that an alcohol extraction method is large in pollution, expensive in equipment and the like are solved, and the application and popularization capacity of the prescription is effectively improved.

Description

Technical Field

[0001] The present invention relates to the field of processing of traditional Chinese medicine compositions, and in particular to a compound traditional Chinese medicine composition for improving the immunity of poultry and a preparation method of the composition. Background Art

[0002] In Traditional Chinese Medicine (TCM), the core of "supporting the body's vital energy and dispelling pathogenic factors" (or "strengthening the body's vital energy and strengthening its foundation") lies in supporting the body's vital energy (i.e., immunity). Existing immune-boosting Chinese medicines primarily use Qi-tonifying and Yang-raising herbs like Astragalus and Codonopsis. While these herbs effectively replenish Qi, activate blood circulation, and enhance immunity, they often suffer from over-tonification and excessive Yang Qi, requiring frequent adjustments to prescriptions and dosages. Furthermore, individual differences often require customized prescriptions.

[0003] For animals, it is difficult to do the same as humans: observe, smell, ask, and feel, and make fine adjustments to the prescription. Moreover, animal farming is often large-scale farming, and the immunity levels of different animal individuals are greatly affected by genetic factors, breeding conditions, and stress factors. In the application of Chinese medicine compositions in the breeding field, it was found that many prescriptions in the prior art were directly transferred from humans to the field of poultry farming. However, the physiological and anatomical structure of poultry is different from that of humans (poultry has a short intestine and a short food residence time). Therefore, the direct transfer of the prescription will cause the immunity enhancement effect to fail to achieve the expected technical problem due to the excessive use of tonic medicines and the neglect of the actual absorption capacity of poultry.

[0004] To enhance the universality of prescriptions in poultry farming and overcome individual differences in immunity and absorption challenges, existing solutions need to be improved. Based on the anatomical characteristics of the short digestive tract of poultry, by scientifically combining digestive drugs in tonic prescriptions, an immune-enhancing prescription that aligns with the physiological characteristics of poultry can be constructed. This prescription can promote drug absorption, balance individual drug sensitivity, and reduce the discreteness of individual immune responses, thereby addressing the problem of drug efficacy loss and achieving group immunity enhancement, providing a practical technical solution for large-scale poultry farming.

[0005] During the early stage of research on animal prescriptions, the research team found that the existing Chinese medicine prescriptions for poultry immunity mostly use powerful drugs for replenishing qi and strengthening the spleen, but ignore the technical defects that excessive replenishment leads to poor absorption and the immunity enhancement effect does not achieve the expected effect. For example, the Chinese medicine prescription in comparative document 1: CN 106344645 B uses astragalus as the main medicine, and the Chinese medicine prescription in comparative document 2: CN101249157B uses astragalus as the main medicine. Both of them mainly use astragalus to replenish qi and lack digestive medicine; comparative document 3: Although the Chinese medicine prescription in CN 118320021A also uses the same medicinal materials such as astragalus and polygonatum as the improved prescription in this application, the prescription still mainly uses astragalus to replenish qi and polygonatum as an adjuvant, and the prescription does not have digestive Chinese medicine that can relieve excessive replenishment. It can be seen that existing prescriptions for enhancing immunity are mostly based on "tonification" and do not address the "spleen and stomach stagnation problem" caused by excessive tonification. Generally speaking, strengthening the spleen is a common technical approach for compound prescriptions that enhance immunity. However, common digestive ingredients often inhibit the "spleen-tonifying" function. This mutual balance between medicinal properties requires special attention to correct medication and reasonable dosage when formulating prescriptions. Only in this way can the synergistic consideration of spleen-tonifying and digestive functions be achieved, effectively improving immune function, avoiding the imbalance of yin and yang caused by excessive tonification, and preventing spleen and stomach stagnation from affecting drug absorption, thereby making the prescription more comprehensive and adaptable in clinical application.

[0006] In addition, in order to obtain polysaccharide components with medicinal effects, the existing technology often uses an extraction process to enrich traditional Chinese medicine polysaccharides. In the existing technology, the most commonly used method for extracting polysaccharides is the ethanol extraction process. However, the ethanol extraction process has the following characteristics: high raw material and energy consumption costs, complex processes, high raw material cost pressure during large-scale production, and ethanol extraction is prone to environmental pollution. There are also technical problems such as compatibility defects with immunomodulatory components, which will cause the precipitation of water-soluble polysaccharides in traditional Chinese medicines that improve immunity, resulting in a decrease in polysaccharide extraction rate. Therefore, in order to effectively improve the universality of prescriptions in poultry farming and overcome the defects of short poultry digestive tracts, poor absorption, and large differences in individual immunity, it is necessary for us to improve the existing prescriptions related to improving immunity, with the purpose of promoting absorption and reducing individual dispersion, and developing suitable prescriptions with "tonifying + digestion-promoting" as the core compatibility that can be used to improve group immunity. At the same time, in order to strengthen the extraction of traditional Chinese medicine polysaccharide components, it is necessary to improve the extraction process and develop a new path that does not rely on ethanol extraction. Summary of the Invention

[0007] In light of the above, to effectively improve the universality of prescriptions, overcome individual differences in immunity, and effectively overcome the "spleen and stomach stagnation problem" caused by excessive tonics, we need to conduct experimental research on prescriptions, focusing on tonifying Qi and supplemented by promoting digestion. This approach, while strengthening the spleen, can also help eliminate stagnation through digestion, thereby achieving a dynamic balance in the body's Qi and blood circulation. By reducing individual dispersion, we can develop suitable prescriptions that can be applied to improve group immunity rather than individual immunity. At the same time, to enhance the extraction of polysaccharides from traditional Chinese medicines, we need to improve the extraction process and develop new methods for extracting polysaccharides from traditional Chinese medicines that do not rely on ethanol.

[0008] A compound Chinese medicine composition for improving the immunity of poultry consists of 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel, 10-15 parts of hawthorn, 5-8 parts of black plum and 2-5 parts of liquorice.

[0009] Furthermore, the compound Chinese medicine composition consists of 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel, 10 parts of hawthorn, 5 parts of black plum and 2 parts of liquorice.

[0010] Furthermore, the preparation method of the compound Chinese medicine composition is as follows: after crushing astragalus, red dates, poria, white atractylodes, tangerine peel, hawthorn, black plum, and licorice, add 12 times of water and soak for 30 minutes, decoct for 1.5 hours, filter and then add 12 times of water and decoct for 1.5 hours, decoct for a total of 3 times, combine the 3 decoctions, and concentrate by rotary evaporation to prepare an oral solution with a concentration of 0.5 g / mL containing the crude drug.

[0011] Furthermore, when the compound Chinese medicine composition is used, the Chinese medicine oral liquid is diluted with water to a Chinese medicine liquid with a volume percentage of 2.4%.

[0012] Furthermore, hawthorn and black plum are reduced from the compound Chinese medicine composition to obtain compound three, and the prescription composition of compound three is as follows: 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel and 2-5 parts of liquorice.

[0013] Furthermore, hawthorn and black plum were reduced from the compound Chinese medicine composition to obtain compound three, and the prescription composition of compound three was as follows: 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel and 2 parts of liquorice.

[0014] Furthermore, hawthorn and black plum are replaced in the compound Chinese medicine composition to obtain Compound IV, and the prescription composition of Compound IV is: 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel, 10-15 parts of polygonatum, 10-15 parts of malt and 2-5 parts of licorice.

[0015] Furthermore, hawthorn and black plum were replaced in the compound Chinese medicine composition to obtain Compound IV, and the prescription composition of Compound IV was as follows: 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel, 10 parts of polygonatum, 10 parts of malt and 2 parts of licorice.

[0016] Furthermore, two herbs, polygonatum and malt, are added to the compound Chinese medicine composition to obtain Compound Five. The prescription composition of Compound Five is as follows: 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel, 10-15 parts of hawthorn, 5-8 parts of black plum, 2-5 parts of liquorice, 10-15 parts of polygonatum and 10-15 parts of malt.

[0017] Furthermore, two herbs, polygonatum and malt, are added to the compound Chinese medicine composition to obtain Compound Five. The prescription composition of Compound Five is as follows: 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel, 10 parts of hawthorn, 5 parts of black plum, 2 parts of licorice, 10 parts of polygonatum and 10 parts of malt.

[0018] The present invention also includes application of the compound Chinese medicine composition in improving the bursa index of chicken.

[0019] The present invention also includes the use of the compound Chinese medicine composition in improving the spleen organ index, liver organ index and / or serum IgG level of chickens.

[0020] The present invention also includes the application of the compound three in increasing the body weight of chickens.

[0021] The present invention also includes the use of the compound five in the preparation of vaccine immune adjuvants and / or immune enhancers.

[0022] Furthermore, the dosage of the compound five is 1.6%.

[0023] The present invention also includes a preparation method for the compound Chinese medicine composition, which comprises the following steps: weighing each medicinal material according to the number of parts of the compound Chinese medicine composition, crushing the medicinal materials with a grinder, soaking the medicinal materials with 12 times the amount of water for 30 minutes, and then decocting the medicinal materials for 1.5 hours; filtering out the medicinal liquid, adding 12 times the amount of water to the medicinal residues, and decocting the medicinal materials for 1.5 hours; extracting the medicinal materials three times; combining the three extracts, and concentrating the extracts to prepare an oral liquid with a concentration of 0.5 g / mL or a granular preparation.

[0024] The present invention has the following beneficial effects:

[0025] 1. The basic prescription of this application uses Astragalus membranaceus to invigorate Qi as the main medicine, Atractylodes macrocephala to invigorate the spleen and dry dampness, red dates to nourish blood and calm the mind, and Poria cocos to invigorate the spleen and eliminate dampness as the ministers, enhancing the spleen and lung function of Astragalus membranaceus, and supplemented with tangerine peel, hawthorn, black plum and other digestive and intestinal astringent drugs to assist the main and minister drugs in regulating Qi, digesting food and removing stagnation, and playing the role of digestion and eliminating accumulation while invigorating Qi and invigorating the spleen. Animal experiments have shown that the basic prescription has a good effect on promoting the development of Fabricius bursa, and the high-dose group is significantly better than the commonly used drug Astragalus polysaccharide, which is a rare prescription for improving immunity.

[0026] 2. Based on the basic formula, the research team conducted studies on the basic formula by adding, reducing, and replacing ingredients. They found that for the basic formula, if chicken gizzard lining and white lentils, which have the function of strengthening the spleen and digesting food, are added, the liver index can be significantly improved, thereby improving the immune function; or if hawthorn and black plum, which have the function of digesting food, are reduced, the immune function and growth can be significantly improved by significantly improving the spleen index. This shows that in the basic formula, both adding and reducing digestive drugs are likely to improve the immune function. The combination of Chinese medicines is not a simple addition or subtraction, and the synergistic effects of various drugs should be considered.

[0027] 3. In improving the basic formula, we found that the immune-boosting effect was even more pronounced if the dual Qi-tonifying and Yin-nourishing herbs of "Astragalus and Polygonatum" were used instead. In experiments with Compounds 4 and 5, we found that Compounds 4 and 5 significantly increased IgG levels compared to the basic formula. Compound 4 replaced the basic formula's black plum and hawthorn with Polygonatum and malt, while Compound 5 added hawthorn and black plum. In Compound Five, Polygonatum sibiricum strengthens the depth of nourishment from the perspective of replenishing Qi and nourishing Yin. The added malt is good at eliminating various food stagnations such as rice, flour, meat and eggs. After being combined with Hawthorn, it not only enhances the elimination of greasy meat stagnation, but also expands the scope of elimination of rice and flour stagnation, and is supplemented with Tangerine Peel to regulate Qi, so as to promote circulation and nourishment, so that the prescription has the effects of "strengthening the spleen + replenishing Qi and nourishing Yin + digestion". The final test found that the IgG value of Compound Five reached the highest and the immune function was the best. Moreover, compared with other experimental groups, the standard deviation was smaller, indicating that the difference between individual animals was the smallest and it was most capable of achieving the improvement of group immunity. In Compound Five (added recipe), Hawthorn and Black Plum had a significant synergistic effect on the efficacy of other drugs, but in Compound Three (reduced recipe), after the removal of Hawthorn and Black Plum, the IgG value was significantly higher than that of the basic recipe, indicating that Hawthorn and Black Plum had a synergistic effect in the "Astragalus + Polygonatum sibiricum" dual-drug combination. This finding also reflects the importance of the synergy of various drugs in the Chinese medicine composition. The immune enhancement test of chicken bursal vaccine further proved the significant immune enhancement effect of compound five. Drinking water with 1.6% compound five can significantly enhance the immune effect of bursal vaccine, and is significantly better than astragalus polysaccharide in improving antibody levels and sustained effects.

[0028] 4. In addition, the present application has also developed a pharmaceutical process for extracting Chinese medicine polysaccharides using water extraction, which greatly optimizes the extraction process of Chinese medicine polysaccharides, overcomes technical problems such as environmental pollution, low extraction rate, complex equipment, and separation difficulties in ethanol extraction, and improves the application scope of Chinese medicine. The above-mentioned medicines are used in common under the ratio of the present invention, and are made into Chinese medicine granules and oral liquid preparations through optimized technology, which greatly improves the application of the prescription. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The morphological and size diagrams of the bursa of Fabricius of the blank group, astragalus polysaccharide group and basic formula group in the third dosing cycle (21 days of age); from left to right: blank group, astragalus polysaccharide group, basic formula high-dose group, basic formula medium-dose group, and basic formula low-dose group.

[0030] Figure 2 These are the microscopic images of the bursa of Fabricius in the control group, the high-dose basic prescription group, and the astragalus polysaccharide group.

[0031] Figure 3 This is a graph showing the effects of each formula on T-SOD in the serum of chicks.

[0032] Figure 4 The graph shows the effects of each formula on serum MDA in chicks.

[0033] Figure 5 The results of the effects of each formula on the serum IgG of chicks.

[0034] Figure 6 This is the standard curve of glucose. DETAILED DESCRIPTION

[0035] The technical solution of the present invention is further described below with reference to specific embodiments, but should not be construed as limiting the present invention:

[0036] Example 1

[0037] This embodiment is based on the research of the party, and the details are as follows:

[0038] 1. The basic formula of this application is the "Qizhaoshuling Decoction" developed by the research team: 15 parts of Astragalus, 10 parts of Atractylodes, 20 parts of red dates, 10 parts of Poria, 10 parts of Tangerine Peel, 10 parts of Hawthorn, 5 parts of Black Plum and 2 parts of Licorice.

[0039] In this formula, the monarch herb is Astragalus membranaceus, which strengthens Qi and strengthens the exterior, elevates Yang and lifts the sunken, and enhances the body's immunity against external pathogens. The assistant herb Atractylodes macrocephala strengthens the spleen and eliminates dampness, helping Astragalus membranaceus replenish Qi. Red dates replenish the middle Qi, nourish the blood, and calm the mind. Poria cocos strengthens the spleen and eliminates dampness, helping Atractylodes membranaceus strengthen the spleen. The adjuvant herbs are Tangerine peel, Hawthorn fruit, and Black Plum. Tangerine peel regulates Qi and strengthens the spleen, Hawthorn fruit helps digestion and resolves stagnation, and Black Plum fruit astringes the lungs and astringes the intestines, helping the monarch and assistant herbs regulate Qi, digestion, and relieve stagnation, while also preventing the herbs from being too warm and dry. The assistant herb is Licorice root, which harmonizes the herbs and enhances their synergy.

[0040] The aforementioned herbs, when combined in the present invention's proportions, are decocted and their active ingredients extracted to create a traditional Chinese medicine that significantly improves the body's disease resistance and promotes the development of immune organs (such as the bursa of Fabricius). This formula not only leverages the immune-boosting effects of Qi-tonifying herbs, but also utilizes the auxiliary and adjuvant herbs to dissipate dampness and aid digestion, forming a synergistic mechanism of "tonifying and strengthening the body, while promoting digestion and promoting digestion," ultimately boosting immunity.

[0041] To verify the feasibility of this prescription, the research team used their own "Qizhaoshuling Decoction" (hereinafter referred to as the basic prescription) to conduct experiments on the development of the bursa of Fabricius in chicks. The details are as follows:

[0042] 1. Experimental Animals: 150 one-day-old Hy-Line Grey laying hens were purchased from Xingnong Poultry Farm, Jinxian, Jiangxi Province.

[0043] 2. Extraction process of the basic formula: Weigh astragalus, red dates, poria, atractylodes, tangerine peel, hawthorn, black plum, and liquorice by weight, grind them, add 12 times the amount of water, soak for 30 minutes, decoct for 1.5 hours, filter, add 12 times the amount of water and decoct for 1.5 hours, decoct for a total of 3 times, combine the 3 decoctions, and concentrate them by rotary evaporation to prepare an oral solution with a concentration of 0.5 g / mL containing the crude drug. Before use, dilute the oral solution according to the volume percentage of the dosage treatment in Table 1.

[0044] 3. Feeding method: One-day-old Hailan white egg chicks were raised and managed under the same conditions according to conventional methods. After 3 days of adaptive feeding, they were randomly divided into 5 groups according to body weight, with 3 replicates in each group and 10 chickens in each replicate. Group 1 was the blank control group, Group 2 was the positive control drug Astragalus polysaccharide group, and Groups 3 to 4 were high-, medium-, and low-dose groups of compound Chinese medicine. Each group was given the drug according to Table 1 for 8-9 hours during the daytime during the chickens' activity period, and the drug was replaced with clean water for free drinking after the end of the drug administration. The test cycle was set as follows: 7 days as a dosing cycle, no drug was given for the first 3 days of each cycle, and the drug was given for 4 consecutive days starting from the 4th day; 4 complete dosing cycles were carried out in sequence, and the cumulative test time was 28 days.

[0045] Table 1 Experimental animal groups and treatments

[0046]

[0047] 4. Test indicators and testing methods:

[0048] 1. Body Weight, Feed Intake, and Feed-to-Growth Ratio: Weigh any remaining feed at 9:00 AM each day and add a certain amount of feed to calculate daily feed intake. Weigh the chicks on an empty stomach at 2:00 PM on Day 28. Calculate the average daily gain (ADG), average daily feed intake (ADFI), and feed-to-growth ratio for each group. Record the chicks' initial and final weights (g). Calculate the total feed intake and feed-to-growth ratio during the trial.

[0049] 2. Immune organ index: At 21 and 28 days of age, 2 chickens were taken from each replicate group, for a total of 6 chickens in each group. Blood was collected from the heart, and the chickens were sacrificed by exsanguination. The bursa of Fabricius and spleen were weighed. The index was calculated using the following formula:

[0050] Immune organ index (mg / g) = immune organ weight (mg) / body weight (g).

[0051] 3. IgG antibody level: At 21 and 28 days of age, two chickens were randomly selected from each replicate group, for a total of 6 chickens in each group. They were weighed and numbered, and blood was collected from the heart. After the serum was naturally precipitated, it was centrifuged and dispensed into EP tubes. The serum was stored in a -80℃ refrigerator until further use for the detection of IgG antibody levels.

[0052] 4. Microscopic observation of bursal tissue.

[0053] 5. Data Processing: SPSS 26.0 software was used for data processing and statistical analysis. Means and standard deviations for each group were calculated using the one-way ANOVA procedure. Statistical analysis of the experimental data was performed using the least significant difference (LSD) method and Duncan's multiple comparisons. All data are presented as mean ± standard deviation (mean ± SD).

[0054] 5. Results and Analysis

[0055] 1. The effects of each formula on chicken growth performance and immune organ index are shown in Table 2, Table 3 and Figure 1-Figure 2 As shown:

[0056] Table 2 Average body weight and average daily weight gain of chickens in each group (g)

[0057]

[0058]

[0059] As shown in Table 2, although there was no significant difference in the weight of the chickens in the high-dose basic formula group at 28 days of age compared with the blank group, the average daily weight gain of each chicken in the basic formula group was 3.55% higher than that in the blank group. This indicates that the high-dose basic formula has a certain growth-promoting effect while improving immunity.

[0060] Table 3 Comparison of immune organ indexes in each group (mg / g)

[0061]

[0062] Note: Data in the table with a ** indicates an extremely significant difference compared to the blank control group (p < 0.01), and * indicates a significant difference (p < 0.05). ## indicates an extremely significant difference compared to the Astragalus polysaccharide group (p < 0.01), and # indicates a significant difference (p < 0.05). No mark indicates a non-significant difference (p > 0.05). Same as the following table.

[0063] The bursa of Fabricius is a central immune organ and a key site for cell growth and differentiation in the avian antibody production system. It produces large numbers of plasma cells and various specific B cells (bursa-dependent lymphocytes). Table 3 shows that at 21 days of age, the astragalus polysaccharide group and the high-dose basic formula group were significantly higher than the blank group (p < 0.05). This indicates that high-dose "Qizao Shuling Decoction" and astragalus polysaccharide can both increase the bursal index. At 28 days of age, the basic bursal index in the high-dose group was significantly higher than that in the blank control group and the other drug-treated groups (p < 0.01). There was no significant difference in bursal index between the blank group and the other drug-treated groups (p > 0.05). The decrease in bursal index in the blank group and the other drug-treated groups is due to the rapid weight gain of the birds during this period, while the bursal index did not significantly increase. In contrast, the bursal index in the high-dose group continued to increase with weight gain. The spleen is a peripheral immune organ. There was no significant difference in the spleen index of each group at 21 days of age. The spleen index of the high-dose basic formula group at 28 days of age was significantly higher than that of the blank control group. Although the astragalus polysaccharide was higher than that of the blank control group, there was no significant difference.

[0064] 2. Effects of each formula on chicken bursa tissue Figure 1 and Figure 2 shown.

[0065] in, Figure 1 The comparison photos of the bursa sizes of each group on the 28th day are shown. Figure 2 HE staining results, from Figure 1 It can be seen that the bursa of Fabricius in the high-dose basic prescription group was significantly larger than that in the other groups.

[0066] Figure 2 Paraffin sections of the bursa of Fabricius from the blank group, the APS group, and the high-dose basic formula group were prepared and stained with HE. As shown in the figure, the demarcation between the cortex and medulla in the bursal nodules in the high-dose and APS groups was more distinct than in the blank group, indicating that the drug promoted the orderly differentiation of lymphoid tissue within the bursa and enhanced its development. Cellular analysis revealed a higher density of lymphocytes in the cortex of the high-dose and APS groups than in the blank group, suggesting that the drug can promote lymphocyte proliferation. The high-dose group stained darker in both the cortex and medulla than in the APS and blank groups.

[0067] 3. The effects of each formula on chicken immunoglobulin G (IgG) levels are shown in Table 4:

[0068] Table 4 Effects of each group on chicken serum IgG (ug / mL)

[0069]

[0070] As shown in Table 4, the immunoglobulin G (IgG) levels in the serum of chickens treated with the APS and high-dose groups were significantly higher than those in the blank control group at 21 days of age (p < 0.05), with no significant difference between the two groups. This indicates that both groups can effectively activate humoral immunity at this stage. While the IgG levels in the medium-dose and low-dose groups of the basic formula increased numerically, they did not reach statistical significance, indicating a lack of significant immune-enhancing effects. At 28 days of age, the IgG level in the APS group decreased compared to that in the 21-day group, while the IgG levels in the high-, medium-, and low-dose groups of the basic formula continued to increase. The high-dose group was significantly higher than the blank control group (p < 0.01), increasing by 120.8% compared to the blank group, significantly higher than the APS group (p < 0.05), and 65.67% higher than the APS group. The immune-enhancing effects of the medium- and low-dose groups did not reach significance (the low-dose group had large standard deviations and heterogeneous variances, resulting in no significant differences), highlighting the long-term immune enhancement advantage of the high-dose group.

[0071] In this embodiment, the basic prescription consists of 15-20 parts of Astragalus, 10-15 parts of Atractylodes, 15-20 parts of red dates, 10-15 parts of Poria, 10-15 parts of dried tangerine peel, 10-15 parts of hawthorn, 5-8 parts of black plum and 2-5 parts of licorice, which can achieve the effect. Within this dosage range: the bursal index of the chicken on the 21st day reached 4.25-6.39 mg / g, which was significantly higher than that of the blank group, and the bursal index of the chicken on the 28th day reached 4.04-6.30 mg / g, which was significantly higher than that of the blank group; the serum IgG of the chickens in the high-dose group of the basic prescription on the 21st day reached 124.04-207.80 ug / mL, which was significantly higher than that of the blank group, and the serum IgG of the chickens on the 28th day reached 155.56-212.02 ug / mL, which was significantly higher than that of the blank group.

[0072] Example 2:

[0073] This example is an improvement to Example 1, "Qizao Shuling Decoction" (hereinafter referred to as the basic formula). Our previous animal experiments found that the basic formula has shown significant effects in promoting the development of poultry immune organs and increasing IgG compared to astragalus polysaccharides. However, in order to further optimize its clinical application value and reduce the dosage and cost of medication, the research team conducted the following treatments on the prescription: adding flavors, reducing flavors, and replacing flavors. The following four prescriptions were selected from a large number of prescriptions and relevant animal experiments were conducted, as follows:

[0074] ① Compound Recipe 2 (Added Recipe 1): Add chicken's gizzard lining and white lentils to the basic recipe, so that the final recipe is: 15 parts of Astragalus, 10 parts of Atractylodes, 20 parts of red dates, 10 parts of Poria, 10 parts of dried tangerine peel, 10 parts of hawthorn, 5 parts of black plum, 2 parts of licorice, 10 parts of chicken's gizzard lining and 10 parts of white lentils.

[0075] Two new herbs have been added to this prescription: chicken's gizzard lining and white lentil. Chicken's gizzard lining is sweet and neutral in nature, entering the spleen, stomach, small intestine, and bladder meridians. It has stomach-strengthening and digestive properties, containing gastrin and digestive enzymes (effective for relieving food stagnation such as rice, flour, meat, and eggs). Combined with hawthorn, it not only enhances digestion of greasy food but also broadens the scope of digestion for rice and flour-based foods. White lentil, on the other hand, strengthens the spleen and eliminates dampness without damaging the stomach, relieves summer heat and detoxifies, and also boosts immunity. It enters the spleen and stomach meridians, is sweet and slightly warm in nature. Both herbs primarily strengthen the spleen and digest food, playing a role in the prescription's digestive function.

[0076] ② Compound Recipe 3 (Reduced Flavor Recipe): Subtract hawthorn and black plum from the basic recipe, so the final recipe is: 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel and 2 parts of liquorice.

[0077] Two herbs omitted from this prescription: hawthorn and black plum. Hawthorn is sour and sweet, slightly warm, and enters the spleen, stomach, and liver meridians, promoting digestion and strengthening the stomach (it's good at dissipating greasy food and meat stagnation). Black plum is sour and astringent, neutral in nature, and enters the liver, spleen, lung, and large intestine meridians. Its primary functions are to astringe yin and promote fluid production, as well as to astringe the intestines. In this prescription, hawthorn primarily serves as a digestive aid, while black plum serves as an astringent. To verify the core "digestive and yin-aggregating" functions of hawthorn and black plum in the prescription and their necessity for compatibility with other herbs, a design of flavor reduction or substitution was used to accurately determine whether these two herbs are essential to the prescription, providing a basis for determining the optimal formula.

[0078] ③ Compound Prescription 4 (replacement prescription): Replace the hawthorn and black plum in the basic prescription with polygonatum and malt respectively, that is, the final prescription is: 15 parts of astragalus, 10 parts of white atractylodes, 20 parts of red dates, 10 parts of poria, 10 parts of tangerine peel, 10 parts of polygonatum, 10 parts of malt and 2 parts of liquorice.

[0079] The two herbs replaced in this prescription are: Polygonatum odoratum is sweet and neutral in nature. It enters the spleen, lung, and kidney meridians, and has the effects of nourishing yin and moistening the lungs, tonifying the spleen and replenishing qi, and nourishing the kidneys and replenishing essence. It has the characteristics of "tonifying both qi and yin, and harmonizing the lungs, spleen, and kidneys." Malt is sweet and neutral in nature, and enters the spleen and stomach meridians. It has the effects of "sweet and neutral digestion, soothing the liver and regulating qi." Malt has a strong and comprehensive digestive effect (it is good at digesting various food stagnations such as rice, flour, meat, and eggs). This prescription focuses on tonifying qi, nourishing yin, and promoting digestion rather than astringency.

[0080] ④ Compound Five (Added Formula 2): Add Polygonatum sibiricum and malt to the basic formula, that is, the final formula is: 15 parts of Astragalus membranaceus, 10 parts of Polygonatum sibiricum, 10 parts of Atractylodes macrocephala, 20 parts of red dates, 10 parts of Poria cocos, 10 parts of dried tangerine peel, 10 parts of hawthorn, 10 parts of malt, 5 parts of black plum and 2 parts of liquorice.

[0081] The most significant difference between this modified formula and Compound 2 is that it utilizes a dual-principle formula, with Polygonatum sibiricum and Astragalus membranaceus serving as the principal ingredients. Astragalus membranaceus tonifies Qi, while Polygonatum sibiricum nourishes Yin, creating a dual-tonification mechanism for Qi and Yin. Astragalus membranaceus significantly tonifies the spleen, elevating Yang and strengthening the exterior, while Polygonatum sibiricum nourishes Yin and moistens the lungs, tonifying the spleen and benefiting the kidneys, addressing the dual needs of spleen development (Qi) and humoral immunity (Yin). Another significant difference is that Compound 2 incorporates a digestive agent, consisting of hawthorn and chicken's gizzard lining, while Compound 5 incorporates hawthorn and malt. Both chicken's gizzard lining and malt contain proteases and amylases, and are effective in digesting food stagnation, including rice, flour, meat, and eggs. Comparing the results of Compound 2 and Compound 5 further clarified the necessity of including Polygonatum sibiricum and malt. The combined effects of Compound 5 not only strengthen the spleen and eliminate dampness, regulating Qi, but also, through their digestive properties, prevent stagnation caused by tonics, achieving a balanced effect of "tonifying while promoting circulation, tonifying without stagnation." Overall, Compound Five constructs an efficacy system of "strengthening the body's health, strengthening the foundation, and promoting circulation and replenishing the body" through the combined combination of "strengthening the spleen + replenishing qi and nourishing yin + digesting food and relieving stagnation".

[0082] To verify the immune function of the above prescription, the applicant conducted animal experiments, as follows:

[0083] 1. Preparation of prescription: According to the above Chinese medicine formula (basic prescription, compound prescription 2, compound prescription 3, compound prescription 4 and compound prescription 5), grind various medicinal materials, add 12 times water to soak for 30 minutes, decoct for 1.5 hours, decoct for a total of 3 times, combine the 3 decoctions, and concentrate by rotary evaporation to contain 0.5g / mL of crude drug for use.

[0084] 2. Experimental animals, grouping and treatment: After 3 days of adaptive feeding, one-day-old Hailan gray laying chicks were randomly divided into 6 groups according to body weight, with 15 chickens in each group. Group 1 was the blank control group, and groups 2-6 were respectively the basic formula group, compound formula group 2, compound formula group 3, compound formula group 4, and compound formula group 5. The drug was administered for 8-9 hours during the daytime during the chicken's activity period, and the water was replaced after the administration. Each drug-dosing group was administered with drinking water containing 1.6% crude drug (the dosage in the basic formula of Example 1). The experimental cycle design was the same as that of Example 1. During the experiment, the room maintained appropriate temperature, humidity and ventilation, and the sanitation of the chicken house was maintained. All chickens were raised and managed under the same conditions according to conventional methods.

[0085] The reason for selecting the medium dose of the basic formula in Example 1 in this example is that the high dose (2.4%) of the basic formula can significantly increase the IgG level. If the high dose of the basic formula is used, the improved formula may mask the advantages of the improved formula due to effect saturation or data fluctuations. The medium dose (1.6%) can not only verify the optimization effect of the improved formula in increasing IgG levels and reducing standard deviations, but also reduce medication costs to meet actual application needs.

[0086] III. Sample Collection and Processing: At 28 days of age, 10 chickens were randomly selected from each group. After fasting for 12 hours, blood was collected from the heart. Serum was allowed to precipitate naturally and then centrifuged. The serum was aliquoted into EP tubes and stored in a -80°C refrigerator until needed. The chicks were exsanguinated, and the spleen, bursa of Fabricius, and liver were removed. They were rinsed with physiological saline, dried with filter paper, and weighed. Organ indices were calculated, and a portion of the spleen was fixed in 4% paraformaldehyde for preparation of spleen tissue sections.

[0087] 4. Testing indicators and testing methods:

[0088] 1. Effects of Each Formula on Chicken Growth: All chickens were weighed on an empty stomach at the beginning and end of the experiment. Feed intake was recorded daily during the experiment. After the experiment, the average weight, average daily gain, and feed-to-weight ratio of each group were calculated.

[0089] 2. Determination of organ index: When the chicks are 28 days old, 10 chicks are randomly selected from each group, blood is collected from the heart, and the chicks are sacrificed by exsanguination. The spleen, bursa of Fabricius, and liver are weighed. The organ index is calculated according to the following formula:

[0090] Organ index (mg / g) = organ weight (mg) / body weight (g).

[0091] 3. Serum antioxidant index determination: superoxide dismutase (T-SOD) activity, malondialdehyde (MDA) index, strictly follow the kit instructions (Nanjing Jiancheng Bioengineering Institute).

[0092] 4. Immune indicators: Serum immunoglobulin G (IgG) antibody levels were measured using an ELISA kit (Shanghai ELISA Biotechnology Co., Ltd.).

[0093] 5. Data processing: Same as Example 1.

[0094] 6. Results and Analysis

[0095] 1. The effects of each formula on chicken growth performance are shown in Table 5:

[0096] Table 5 Effects of each formula on chicken growth performance

[0097]

[0098] Note: Data in the table with ** indicates extremely significant difference compared with the blank control group (p < 0.01), * indicates significant difference (p < 0.05). ## indicates extremely significant difference compared with the baseline (p < 0.01), # indicates significant difference (p < 0.05), and no mark indicates no significant difference (p > 0.05). Same below.

[0099] As shown in Table 5, in terms of body weight at the end of the experiment, Compound 2 and Compound 3 were significantly higher than the blank group; in terms of average daily weight gain per chicken, the average daily weight gain of the basic formula, Compound 2, Compound 4 and Compound 5 were slightly higher than the blank group, but there was no significant difference in the average daily weight gain of these groups (p>0.05), while the daily weight gain of Compound 3 was significantly higher than that of the blank group and the basic formula ((p<0.01)) and the feed-to-weight ratio was the lowest, indicating that Compound 3 can significantly promote chicken growth and improve feed conversion rate.

[0100] 2. The effects of each formula on chicken organ indexes are shown in Table 6:

[0101] Table 6 Effects of each formula on chicken immune organ index (mg / g)

[0102]

[0103] As shown in Table 6, regarding the basic formula, the basic formula in Example 2, administered at the medium dose (1.6%) of the basic formula in Example 1, showed no significant differences in any organ index compared to the blank control group, consistent with the results of Example 1. However, a detailed analysis of the bursal index of the basic formula revealed that 50% (5 / 10) of the 10 samples had an index exceeding 7.8, with 4 samples exceeding 8.0. In contrast, the bursal index of all 10 samples in the blank control group was below 7.8. This result suggests that the basic formula has a specific effect on promoting bursal development, and while the medium dose (1.6%) can be significantly effective in some sensitive individuals, it cannot be applied to all individuals. Therefore, the recommended clinical dosage for the basic formula is the high dose (2.4%) of Example 1.

[0104] From the perspective of compound 2, the spleen index of compound 2 was significantly higher than that of the basic formula (p < 0.05), the bursa index was slightly higher than that of the blank group and slightly lower than that of the basic formula, but it did not reach a significant level (p > 0.05), and the liver index was extremely significantly higher than that of the blank group (p < 0.01) and significantly higher than that of the basic formula (p < 0.05); this shows that after adding chicken gizzard lining and white lentil, two spleen-strengthening and digestion-promoting medicines, the spleen and stomach function of the chickens was further improved, and the liver-protecting and spleen-strengthening effects of the medicine were more obvious. White lentil strengthens the spleen to help the qi flow and transform, and eliminates dampness to eliminate qi stagnation; chicken gizzard eliminates accumulation and stagnation to promote qi flow, which also indirectly indicates that the chickens have a short digestive tract and a short time to digest and absorb nutrients, and adding digestive medicine can promote absorption.

[0105] From the perspective of compound three, the spleen index of compound three was significantly higher than that of the basic formula group (p < 0.05), the bursa index was higher than that of the blank group and the basic formula, but did not reach a significant level (p > 0.05), and the liver index was significantly higher than that of the blank group (p < 0.05). Hawthorn (digestive) and black plum (astringent) were subtracted from this prescription, and these two medicines mainly play the role of digestion and astringency, which is opposite to the design idea of ​​compound two with added flavors. However, the effect of improving the spleen index and liver index is better than that of the basic formula. This shows that when the basic formula of this application is improved, focusing on digestion (increasing digestive drugs) or reducing digestion (reducing digestive and astringent drugs) can play the role of strengthening the spleen and protecting the liver. This also reflects the synergistic effect of the drugs in the prescription and the unexpected technical effect.

[0106] From the perspective of Compound 4, the spleen index of Compound 4 was significantly higher than that of the basic formula group (p < 0.05), the bursa index was slightly higher than the blank group and lower than the basic formula, but it did not reach a significant level (p > 0.05), and the liver index was higher than the blank group and the basic formula, but there was no significant difference (p > 0.05). Compound 4 uses Polygonatum (Yin nourishing) and Malt (Digestive) to replace Crataegus pinnatifida (Digestive) and Plum (Astringent) in the basic formula. Polygonatum is introduced to enhance the effect of replenishing Qi and nourishing Yin, and Malt is used to soothe the Liver and regulate Qi, forming a synergistic effect of digestion and regulating Qi with Tangerine Peel, promoting the circulation of Qi and blood, and improving the spleen-strengthening effect, but there is no obvious effect on the development of the bursa.

[0107] From the perspective of Compound Five, the spleen organ index of Compound Five was significantly higher than that of the blank group (p < 0.05, an increase of 16.2%), the bursa of Fabricius organ index was slightly higher than that of the blank group and lower than that of the basic formula, but it did not reach a significant level (p > 0.05), and the liver organ index was slightly higher than that of the blank group and the basic formula, but there was no significant difference (p > 0.05). Compound Five added Polygonatum and malt, and the prescription still contained Hawthorn and Black Plum. Polygonatum strengthens the depth of nourishment from the perspective of replenishing Qi and nourishing Yin, and the added malt is good at eliminating various food stagnation such as rice, flour, meat, and eggs. After being combined with Hawthorn, it not only enhances the elimination of greasy meat stagnation, but also expands the scope of elimination of rice and flour stagnation, and is supplemented with Tangerine Peel to regulate Qi and promote tonification. The whole prescription is designed to form a composite effect of "strengthening the spleen + replenishing Qi and nourishing Yin + digestion"; from the experimental results, the spleen-strengthening effect of this prescription has been more significantly improved.

[0108] 3. Effect of drugs on chicken serum IgG level: In humoral immunity, B lymphocytes differentiate into plasma cells under the action of antigens to produce specific antibodies. The level of antibodies directly reflects the resistance of livestock and poultry. The serum IgG antibody level is a key indicator for measuring humoral immune function. The effects of drugs on chicken serum IgG level are shown in Table 7 and Figure 5 As shown:

[0109] Table 7 Effects of each prescription on chicken serum IgG

[0110]

[0111] As shown in Table 7, there was no significant difference in the chicken serum IgG of the basic formula in this example, but there was a significant difference in Example 1, mainly because the high dose (2.4%) was selected in Example 1, while the medium dose group (1.6%) was selected in this example. The other dosing groups also chose the medium dose (1.6%) consistent with the basic formula. The IgG levels of Compound IV and Compound V were extremely significantly higher than those of the blank control group and the basic formula group (p < 0.01); the IgG level of the Compound III group was significantly higher than that of the blank control group and the basic formula group (p < 0.05); the IgG level of the Compound II group was significantly higher than that of the blank control group (p < 0.05); the IgG level of the Compound V was increased by 194.74% compared with the blank group, 131.58% compared with the basic formula, 78.06% compared with Compound II, 45.65% compared with Compound III, and 37.42% compared with Compound IV; this shows that the three improved formulas of the present application, Compound III, Compound IV and Compound V, all significantly or extremely significantly improved the immune function of animals, and are all good formulas for enhancing the immune function of animals.

[0112] From the perspective of the standard deviation within the group: the standard deviation within the blank group, the basic formula, and the compound formula 2 were large, and the data dispersion was high, indicating that the individual differences within the group were prominent, resulting in uneven immunity performance within the group, with large differences; however, the relative standard deviation within the group of compound formula 3, compound formula 4, and compound formula 5 was small, and the data dispersion was low, indicating that the individual differences within the group were not obvious, and the group immunity status was more stable and uniform, indicating that the drug can improve the overall immunity level of the chicken flock.

[0113] Although the IgG value of Compound 2 was significantly better than that of the blank group (p < 0.05), the standard deviation was 32.94, which was at a higher level than that of other experimental groups, indicating that the dispersion of immune responses among groups was high and the group immunity status was not stable enough. Therefore, we did not consider Compound 2 as an effective improvement formula.

[0114] In summary, in addition to achieving the highest immunity, the compound five in this application also has a standard deviation of less than 20 and a coefficient of variation of 12.75%, which effectively solves the technical defect of "uneven immunity" in traditional breeding. Therefore, we prefer compound five as the best improved formula.

[0115] The analysis and comparison of the formula composition, IgG levels, and spleen index of the six groups fully demonstrated the important role of adding Polygonatum sibiricum and Malt to the basic formula, as well as the important synergistic effect of Hawthorn and Black Plum. Polygonatum sibiricum and Malt are both present in Compound Four and Compound Five, and the IgG level of Compound Four is second only to Compound Five. The difference between Compound Four and Compound Five is that Compound Four does not contain Hawthorn and Black Plum, reflecting the important role of Hawthorn and Black Plum in synergizing with other drugs in Compound Five to enhance immunity. Compound Five combines Qi-tonifying (Astragalus membranaceus, Atractylodes macrocephala) with Yin-nourishing (Poria sibiricum, Black Plum) to avoid "warm and dry damage to Yin" or "nourishing and greasy obstruction of the spleen"; it strengthens the spleen and removes dampness (Poria cocos), regulates Qi (tangerine peel), and helps digestion (Hawthorn and Malt) in combination with tonic drugs to achieve "tonifying while promoting circulation, tonifying without stagnation", ultimately significantly improving the body's immune level.

[0116] 4. Effects of each formula on chicken serum MDA and T-SOD Figure 3 and Figure 4 As shown: Among them, Figure 3 The effects of different drug combinations on T-SOD, Figure 4 The effects of different drugs on MDA are shown in Table 1. Figure 3 and Figure 4 As can be seen, serum SOD and MDA levels in the Compound Five, other treatment groups, and the blank group did not reach statistical significance (p>0.05). Although T-SOD in the Compound Five showed an upward trend and MDA showed corresponding changes, neither reached a statistically significant level. This suggests that the primary mechanism of Compound Five's immune-enhancing effects may not rely on antioxidant pathways.

[0117] In this embodiment, the compound three is composed of 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel and 2-5 parts of liquorice, all of which can achieve the desired effect. Within this dosage range: on the 28th day, the chicken serum IgG reached 68.91-127.55ug / mL, which was significantly higher than that of the blank group.

[0118] The compound prescription consisting of 15-20 parts of Astragalus, 10-15 parts of Polygonatum, 10-15 parts of Atractylodes, 15-20 parts of Red Dates, 10-15 parts of Poria, 10-15 parts of Tangerine Peel, 10-15 parts of Malt and 2-5 parts of Licorice can achieve the desired effect. Within this dosage range: on the 28th day, the chicken serum IgG reached 68.91-127.55ug / mL, which was significantly higher than that of the blank group.

[0119] Compound five, which consists of 15-20 parts of Astragalus, 10-15 parts of Polygonatum, 10-15 parts of Atractylodes, 15-20 parts of Red Dates, 10-15 parts of Poria, 10-15 parts of Tangerine Peel, 10-15 parts of Hawthorn, 10-15 parts of Malt, 5-8 parts of Black Plum and 2-5 parts of Licorice, can achieve the desired effect. Within this dosage range: on the 28th day, the chicken serum IgG reached 116.63-153.35ug / mL, which was significantly higher than that of the blank group.

[0120] Example 3:

[0121] To verify the immune-enhancing function of Compound V, the applicant conducted an immune-enhancing experiment on chicken bursal vaccine, as follows:

[0122] 1. Preparation of prescription: Compound Five (modified prescription): 15 parts of Astragalus, 10 parts of Polygonatum, 10 parts of Atractylodes macrocephala, 20 parts of red dates, 10 parts of Poria, 10 parts of Tangerine peel, 10 parts of Hawthorn, 10 parts of Malt, 5 parts of Black Plum and 2 parts of Licorice. Preparation method is the same as Example 2.

[0123] 2. Experimental animals, grouping and treatment: 150 one-day-old Hailan gray egg chicks were randomly divided into 5 groups, with 3 replicates in each group and 10 chickens in each replicate. Group 1 was the vaccine control group (no administration of the simple bursal vaccine), Group 2 was the astragalus polysaccharide positive drug group (astragalus polysaccharide + bursal live vaccine), and Groups 3-5 were the high, medium and low dose groups of Compound Five (Compound Five + bursal live vaccine) in sequence. All chicks were given a bursal live vaccine (Harbin Pharmaceutical Vaccine, production batch number 20240904) once at 14 days of age. The vaccine control group was not administered, and the administration cycle design of the other groups was the same as in Example 2; among them, the crude drug amount of the high-dose group of Compound Five was 1.6% (the same dose as in Example 2), the crude drug amount of the medium-dose group of Compound Five was 0.8%, and the crude drug amount of the low-dose group of Compound Five was 0.4%.

[0124] 3. Sample collection and testing: At 21 and 28 days of age, two chickens were randomly selected from each replicate group, with a total of six chickens in each group. Blood was collected from the chickens' hearts and allowed to stand at room temperature. After the blood was completely coagulated, it was centrifuged at 3500 rpm for 15 minutes to separate the serum. The serum was tested for antibodies using the infectious bursal disease (IBD) antibody detection kit (Shanghai ELISA Biotechnology Co., Ltd.), and the results were judged according to the instructions.

[0125] 4. Data processing: Same as Example 1.

[0126] 5. Results and Analysis

[0127] The effects of each formula on the serum IBD antibody levels of chicks are shown in Table 8:

[0128] Table 8 Effect of Compound Five on IBD Antibody Levels in Chicken Serum (ng / mL)

[0129]

[0130] Note: Data with * indicates significant difference compared with the vaccine control group (p < 0.05), ** indicates extremely significant difference (p < 0.01), # indicates significant difference compared with the astragalus polysaccharide group (p < 0.05), ## indicates extremely significant difference (p < 0.01), and no mark indicates no significant difference (p > 0.05).

[0131] As shown in Table 8, the IBD antibody level in the vaccine control group (basic immunization) was 11.20±1.09 on day 21, but decreased to 8.66±3.44 on day 28, indicating that antibodies naturally decline over time after simple vaccination. Antibody levels in the APS group (positive control, a traditional immune enhancer) were significantly higher on day 21 than in the vaccine control group (p<0.05), demonstrating that APS can enhance the immune response to vaccination. However, while the antibody level at day 28 was slightly higher than in the vaccine control group, the difference was not significant, indicating that APS's "antibody maintenance effect" was limited, with protection declining after 28 days. The antibody level of the high-dose compound five group (1.6%) at 21 days was significantly higher than that of the vaccine control group (p < 0.05), and the antibody level at 28 days was extremely significantly higher than that of the vaccine control group (p < 0.01), and extremely significantly higher than that of the astragalus polysaccharide group (p < 0.01). The results showed that adding 1.6% of compound five to the drinking water of chickens can significantly enhance the body's immune response efficiency to bursal disease vaccine, improve the level of antibody production, play a positive role in the early induction and sustained protection of bursal disease antibodies, and can be used as a vaccine adjuvant or immune enhancer.

[0132] The antibody level in the medium-dose Compound V group at 21 days was significantly higher than that in the vaccine control group, but due to large standard deviations (data dispersion), it did not reach statistical significance. The antibody levels in the low-dose Compound V group at 21 days and 28 days were not significantly different from those in the vaccine group. Therefore, if chickens require enhanced vaccine protection in aquaculture, adding a 1.6% dose of Compound V to drinking water before and after vaccination can not only rapidly increase antibody levels, but also prolong protection and reduce the risk of infection with infectious diseases. This is worthy of trial and promotion in aquaculture.

[0133] Example 4:

[0134] This example is an optimization of the extraction process of a Chinese herbal compound, specifically as follows:

[0135] Polysaccharides from traditional Chinese medicines are the primary active ingredients in traditional Chinese medicines. They possess immunomodulatory properties. Numerous studies have demonstrated their significant impact on improving immune organ indices, stimulating T cell proliferation and activity, regulating cytokine production, and modulating antibody synthesis. Compound Five, composed of ten herbs (astragalus, white atractylodes, red dates, poria, tangerine peel, hawthorn, black plum, liquorice, polygonatum, and malt), is believed to contain polysaccharides that are believed to be effective in enhancing immunity. Due to the numerous ingredients and high dosages required for traditional Chinese medicine formulations, optimizing extraction processes with the goal of achieving high efficiency, reducing dosage, and prioritizing the extraction of active ingredients is a top priority.

[0136] The specific extraction optimization method is as follows:

[0137] 1. Instruments: UV-visible spectrophotometer (Shimadzu Enterprise Management Co., Ltd.), grinder, pipette.

[0138] 2. Materials: Chinese herbal medicine slices including astragalus root, polygonatum, red dates, atractylodes macrocephala, poria, tangerine peel, hawthorn, malt, black plum and liquorice were purchased from Dashenglin Pharmacy.

[0139] 3. Design and validation of extraction methods:

[0140] 1. Drawing of glucose standard curve: Prepare 0.1mg / mL glucose standard solution. Pipette 0.4mL, 0.5mL, 0.6mL, 0.7mL, 0.8mL, 0.9mL, and 1.0mL into test tubes respectively, add double distilled water to make up to 1mL, and then add 4mL of newly prepared anthrone-concentrated sulfuric acid solution (weigh 0.1g anthrone, dissolve in 100mL 80% concentrated sulfuric acid, and mix evenly). Heat the prepared solution in boiling water for 10min, and cool to room temperature. Take the absorbance at 625nm in the ultraviolet spectrophotometer. Draw the standard curve with the glucose content as the horizontal axis and the absorbance as the vertical axis ( Figure 6 ).

[0141] 2. Orthogonal experimental design of extraction process: Weigh the components of the five traditional Chinese medicines in the compound according to the proportion, crush them in a grinder, soak them for 30 minutes, and optimize the extraction process with the polysaccharide content as the index. Based on the single factor experiment, the factors affecting the polysaccharide extraction, such as the number of extractions, extraction time, and water addition times, were analyzed by L9(3 3 )Orthogonal experiment, the orthogonal experiment factor level table is shown in Table 9, and the orthogonal table is shown in Table 10.

[0142] Table 9 Orthogonal test factor level table

[0143]

[0144] 3. Extraction process validation: Take three samples of the prescribed amount of medicinal material and extract them according to the optimal process conditions. Determine the content of crude polysaccharides in each batch of extract.

[0145] 4. Results and Analysis

[0146] (1) Drawing of glucose standard curve, as shown in the following example: Figure 6 As shown, Figure 6 In the graph, the x-axis represents the concentration of the glucose standard solution, and the y-axis represents the absorbance corresponding to the concentration. The equation of the glucose standard concentration-absorbance standard curve is: Y = 6.0214x + 0.0101, r 2 =0.9931. It has a good linear relationship in the range of 0.04-0.1 mg / mL.

[0147] (2) Orthogonal test results: The number of extractions, extraction time, and water addition times were L9(3 3 ) The orthogonal experimental design and results are shown in Table 10.

[0148] Table 10 Orthogonal experimental design table and result analysis

[0149]

[0150]

[0151] In Table 10, the optimal level was selected based on the highest average polysaccharide value k for each factor. The order of priority was determined based on the range of the average values, R. Therefore, the order of priority for the factors influencing polysaccharide extraction was: number of extractions > extraction time > water addition ratio. The optimal combination for polysaccharide extraction was A3 B2 C3: 3 extractions, 1.5 hours, and a water addition ratio of 1:12.

[0152] (3) Results of verification of the repeatability of the extraction process: Based on the optimal extraction process conditions obtained from the orthogonal experiment in Table 10, three sets of parallel experiments were performed to measure the polysaccharide content. The results are shown in Table 11.

[0153] As shown in Table 11, the average polysaccharide content of Compound 5 extracted using the optimal extraction process conditions obtained in Table 10 was 54.64%. The results show that the optimal extraction process produced the highest polysaccharide content. The RSD of polysaccharide content across the three batches was 4.26%, demonstrating good reproducibility (see Table 11), demonstrating the high feasibility and reproducibility of this process.

[0154] Table 11 Repeatability verification results

[0155]

[0156] Therefore, the results of optimizing the preparation process in combination with Table 10 and Table 11 showed that the optimal process for Chinese medicine preparation was as follows: weigh each medicinal material according to Chinese medicine compound five, crush them in a grinder, soak them in 12 times water for 30 minutes, and then boil them for 1.5 hours. After filtering out the medicinal liquid, add 12 times water to the medicinal residue and boil them for 1.5 hours. Extract three times, combine the three extracts, and concentrate them by rotary evaporation to prepare an oral solution with a concentration of 0.5 g / mL containing the crude drug or prepare it into a granular preparation.

[0157] In summary, the basic prescription of this application is based on the idea of ​​tonifying qi and promoting digestion, which can significantly promote the development of the bursa of Fabricius and thus improve the immunity of poultry. In addition, the basic prescription is improved by research on adding flavors, reducing flavors, and replacing prescriptions. The original prescription is optimized to achieve a very significant improvement in immune function. At the same time, it also effectively reduces the standard deviation, narrows the degree of discreteness of data within the group, reduces individual immune differences, and effectively achieves the technical advantage of enhancing group immunity. Adding a 1.6% dose of compound five drinks before and after vaccine immunization can not only quickly increase the level of specific antibodies, but also prolong the vaccine protection time and reduce the risk of disease infection. In addition, this application also optimizes the process and uses water extraction instead of alcohol extraction to make oral liquid or Chinese medicine granule products, which can greatly exert its immune enhancement effect and the effects of strengthening the spleen and nourishing yin, removing dampness and digestion.

[0158] The above description is a detailed description of the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the patent application of the present invention. Any equivalent changes or modifications completed under the technical spirit suggested by the present invention should fall within the patent scope covered by the present invention.

Claims

1. A compound Chinese medicine composition for enhancing the immunity of poultry, characterized in that: The compound Chinese medicine composition consists of 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of dried tangerine peel, 10-15 parts of hawthorn, 5-8 parts of black plum and 2-5 parts of liquorice.

2. The compound Chinese medicine composition according to claim 1, characterized in that The compound Chinese medicine composition is reduced by reducing hawthorn and black plum to obtain compound three, which comprises 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of dried tangerine peel and 2-5 parts of liquorice.

3. The compound Chinese medicine composition according to claim 1, characterized in that The compound Chinese medicine composition is replaced with hawthorn and black plum to obtain Compound IV, which comprises 15-20 parts of astragalus, 10-15 parts of white atractylodes, 15-20 parts of red dates, 10-15 parts of poria, 10-15 parts of tangerine peel, 10-15 parts of polygonatum, 10-15 parts of malt and 2-5 parts of liquorice.

4. The compound Chinese medicine composition according to claim 1, characterized in that Polygonatum sibiricum and malt are further added to the compound Chinese medicine composition to obtain Compound Five, which comprises 15-20 parts of Astragalus membranaceus, 10-15 parts of Atractylodes macrocephala, 15-20 parts of red dates, 10-15 parts of Poria cocos, 10-15 parts of dried tangerine peel, 10-15 parts of hawthorn, 5-8 parts of black plum, 2-5 parts of liquorice, 10-15 parts of Polygonatum sibiricum and 10-15 parts of malt.

5. Use of the compound Chinese medicine composition according to claim 1 in promoting bursal development and improving bursal index.

6. Use of the compound Chinese medicine composition according to any one of claims 1 to 4 in improving spleen organ index, liver organ index and / or serum IgG level.

7. Use of the compound three according to claim 2 in increasing the body weight of chickens.

8. Use of the compound fifth according to claim 4 in the preparation of vaccine adjuvants and / or immunopotentiators.

9. The use according to claim 8, characterized in that The dosage of the compound five is drinking water containing 1.6% of the crude drug.

10. A method for preparing the compound Chinese medicine composition according to any one of claims 1 to 4, characterized in that: The preparation method comprises the following steps: weighing each medicinal material according to the number of portions of the compound Chinese medicine composition, crushing the medicinal material with a grinder, adding 12 times the amount of water to soak for 30 minutes, and then decocting for 1.5 hours; filtering out the medicinal liquid, adding 12 times the amount of water to the medicinal residue and decocting for 1.5 hours; extracting the medicinal material three times; combining the three extracts, and concentrating the extracts to prepare an oral liquid or a granular preparation.

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