Traditional Chinese medicine fermented paste capable of improving production performance and immune performance of laying hens and preparation method of traditional Chinese medicine fermented paste

Abstract

The invention discloses a traditional Chinese medicine fermented paste for improving the production performance and immune performance of laying hens and a preparation method thereof. The traditional Chinese medicine fermented paste is prepared from the following components in parts by weight: 5-30 parts of astragalus membranaceus, 5-30 parts of scutellaria baicalensis, 5-20 parts of radix angelicae, 5-20 parts of bighead atractylodes rhizome, 5-20 parts of honeysuckle and 5-10 parts of herba epimedii. Mixing the traditional Chinese medicine dry powder, adding a fermentation strain, brown sugar and water, fermenting, drying, adding honey and glycerol, and uniformly stirring to obtain the paste. When in use, the traditional Chinese medicine fermented paste is dissolved in drinking water according to the concentration of 1-4g / L. The traditional Chinese medicine fermented paste can effectively improve the production performance and immune performance of laying hens, improve the health condition and enhance the disease resistance, is convenient to use, good in safety and suitable for long-term use, and has high economic value.

Description

Technical Field

[0001] This invention belongs to the technical field of traditional Chinese veterinary medicine preparations and animal feed additives, specifically relating to a fermented traditional Chinese medicine paste for improving the production performance and immune performance of laying hens and its preparation method. Background Technology

[0002] In modern egg-laying hen farming, improving the production performance and immunity of hens is crucial to ensuring profitability and egg quality. While chemically synthesized drugs can enhance immune function in the short term, long-term use may lead to problems such as drug residues, drug resistance, and environmental pollution, and poses potential risks to animal health. In contrast, natural plant extracts offer a relatively safer alternative, possessing various biological activities such as immunomodulation and antioxidant effects. However, natural plant extracts have certain limitations in clinical applications, such as poor palatability and short duration of action, which reduce their practical effectiveness in animal husbandry.

[0003] Probiotics, as a natural microbial additive, have been widely used in animal husbandry in recent years. Probiotic fermentation technology, by introducing specific beneficial microorganisms, can decompose and transform the components of traditional Chinese medicine during fermentation, while simultaneously improving flavor, promoting digestion, and regulating intestinal flora balance. However, existing probiotic fermentation processes have not yet fully utilized the natural active ingredients of traditional Chinese medicinal plants, and their effectiveness in feed still needs further improvement.

[0004] Studies have shown that flavonoids are a class of naturally occurring active ingredients widely found in plants, possessing significant antioxidant, anti-inflammatory, and immunomodulatory functions. Research indicates that flavonoids can effectively scavenge free radicals in the body, reducing the damage caused by oxidative stress; simultaneously, they can regulate immune cell activity, enhance specific and non-specific immune responses, and improve animals' resistance to pathogens. Therefore, increasing the flavonoid content in feed is of significant value in improving the health and immune function of laying hens. Based on this, optimizing the extraction and transformation of traditional Chinese medicine components through innovative probiotic fermentation processes, especially specifically increasing the content of flavonoid active ingredients, while effectively improving the production performance and immunity of laying hens, has become an important technical challenge in modern animal husbandry.

[0005] This invention provides a traditional Chinese medicine fermented paste prepared through an optimized probiotic fermentation process. Using specific probiotic strains and fermentation conditions, the content of flavonoids in the traditional Chinese medicine can be significantly increased, while simultaneously improving the flavor. By optimizing the fermentation process, this fermented paste not only enhances the production performance of laying hens but also effectively strengthens their immunity, achieving an "antibiotic alternative" effect, thereby replacing traditional antibiotics and reducing drug resistance and residue problems. This fermented paste has no side effects, is suitable for long-term use, meets the needs of modern green farming, and has high application value and market prospects. Summary of the Invention

[0006] This invention provides an innovative solution to the problems of drug residues and drug resistance risks associated with traditional chemically synthesized additives in the prior art, as well as the poor palatability and inconvenience of direct application of natural plant extracts.

[0007] Specifically, this invention discloses a traditional Chinese medicine fermented paste for improving the production and immune performance of laying hens and its preparation method. This fermented paste is prepared by optimizing the fermentation of a selected traditional Chinese medicine compound using a specific combination of probiotics. Its core advantage lies in significantly increasing the content of active ingredients in the traditional Chinese medicine (such as flavonoids). The finished product is a paste, easily dissolved in drinking water, making it extremely convenient to use. After feeding laying hens, it can effectively and safely improve their production performance and immune function, exhibiting high efficiency, safety, and ease of application.

[0008] To achieve the above objectives, the present invention adopts the following technical solution: A fermented herbal paste for improving the production and immune performance of laying hens is made from the following components in parts by weight: Astragalus membranaceus 5-30 parts, Scutellaria baicalensis 5-30 parts, Angelica dahurica 5-20 parts, Atractylodes macrocephala 5-20 parts, Lonicera japonica 5-20 parts, and Epimedium brevicornu 5-10 parts.

[0009] Preferably, the fermented herbal paste is made from the following components in parts by weight: 20-25 parts Astragalus membranaceus, 20-30 parts Scutellaria baicalensis, 15-20 parts Angelica dahurica, 10-20 parts Atractylodes macrocephala, 10-15 parts Lonicera japonica, and 5-10 parts Epimedium.

[0010] More preferably, the fermented herbal paste is made from the following components in parts by weight: 20 parts Astragalus membranaceus, 30 parts Scutellaria baicalensis, 15 parts Angelica dahurica, 10 parts Atractylodes macrocephala, 15 parts Lonicera japonica, and 10 parts Epimedium brevicornu.

[0011] The preparation method of the above-mentioned fermented Chinese herbal medicine paste includes the following steps: (1) Mix the above-mentioned dried Chinese medicine powders in a certain proportion to obtain a mixed Chinese medicine powder; (2) Add a compound fermentation agent activation solution composed of compound fermentation agent, brown sugar and water to the mixed powder of Chinese medicine; the mass ratio of the compound fermentation agent, brown sugar and water is 1:1:18. (3) Add water to the mixture obtained in step (2) and ferment it. The fermentation temperature is 27~42℃ and the fermentation time is 1~4 days. Dry it into a paste.

[0012] Preferably, the ratio of the amount of water added to the total mass of the mixed Chinese medicine powder is 1:1.

[0013] Preferably, the amount of the compound fermentation agent activation liquid added is 2-16% of the total mass of the mixed Chinese medicine powder, and the amount of the compound fermentation agent is 1-8‰ of the total mass of the mixed Chinese medicine powder. More preferably, the amount of the activation liquid added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of the compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0014] Preferably, the compound fermentation agent is a combination of Saccharomyces cerevisiae, Lactobacillus plantarum and Bacillus subtilis, and the ratio of the fermentation strains is 1:1:1, 1:2:1, 1:1:2 or 2:1:1, more preferably 1:1:2.

[0015] Preferably, the drying in step (3) involves drying the fermented material at 48°C for 24 hours.

[0016] Preferably, the paste obtained in step (3) above is mixed with 1-5 parts honey and 1-5 parts glycerin by weight, and stirred thoroughly. More preferably, it is 3 parts honey and 3 parts glycerin.

[0017] The above-mentioned fermented Chinese medicine paste can be dissolved in drinking water and fed to laying hens at a concentration of 1-4 g / L. When using it, it can be dissolved in the drinking water of laying hens and allowed to drink freely.

[0018] The aforementioned fermented herbal extract can significantly improve the production and immune performance of laying hens. Compared with traditional Chinese medicine, this invention significantly increases the content of flavonoids through optimized fermentation process, with no obvious side effects, making it suitable for long-term use and exhibiting high safety. Therefore, it can be applied in the preparation of feed additives to improve the production and immune performance of laying hens. The fermented herbal extract improves the production performance of laying hens by promoting the onset of egg production and improving the pathological state of the ovaries and liver. It improves the immune performance of laying hens by effectively reducing the dispersion of antibody titers against various avian influenza viruses and Newcastle disease viruses.

[0019] The advantages of this invention compared with the prior art are as follows: (1) This invention selects six Chinese herbs: Astragalus membranaceus, Scutellaria baicalensis, Angelica dahurica, Atractylodes macrocephala, Lonicera japonica, and Epimedium. Astragalus membranaceus tonifies Qi and strengthens the exterior, Scutellaria baicalensis clears heat and dries dampness, Angelica dahurica dispels wind and eliminates dampness, Atractylodes macrocephala strengthens the spleen and benefits Qi, Lonicera japonica clears heat and detoxifies, and Epimedium tonifies the kidney and strengthens Yang. The effects are synergistic, the formula is reasonable, and it takes into account multiple effects such as tonifying, clearing heat, strengthening the spleen and tonifying the kidney. The synergistic effect improves the physical condition of laying hens, enhances disease resistance and promotes production performance.

[0020] (2) By using a specific combination of probiotics (Saccharomyces cerevisiae, Lactobacillus plantarum, Bacillus subtilis) and optimizing key parameters such as fermentation time, temperature, bacterial dosage and ratio, the traditional Chinese medicine compound is fermented and transformed. This process can significantly increase the content of active ingredients such as flavonoids in the compound, degrade macromolecules into smaller molecules that are more easily absorbed, optimize the fermentation process, and significantly improve the efficacy.

[0021] (3) After fermentation, the product is dried at low temperature and then mixed with honey and glycerin to make a paste. It is easy to dissolve in water and convenient to use. The addition of honey and glycerin not only improves the physical properties of the preparation, but also enhances its palatability, making it more acceptable to laying hens.

[0022] (4) The product of this invention is made from pure Chinese medicine fermented with probiotics. It has no chemical drug residues or drug resistance issues, no obvious side effects, and is suitable for long-term use by laying hens. It can effectively improve the production performance (such as egg production rate and egg quality) and immune performance (such as antibody level and immune organ index) of laying hens, enhance disease resistance, and conform to the development trend of green, healthy and antibiotic-free breeding in modern animal husbandry. It has high safety and economic value. Attached Figure Description

[0023] Figure 1 Effects of fermented Chinese herbal medicine extract on the morphology and structure of laying hen livers (A) Liver image stained with H&E in the blank control group; (B) Enlarged view of a local liver stained with H&E in the blank control group; (C) Liver image stained with H&E in the group with 2 g / L fermented paste; (D) Enlarged view of a local liver stained with H&E in the group with 2 g / L fermented paste.

[0024] Figure 2 Effects of fermented Chinese herbal medicine paste on the morphology and structure of laying hen ovaries (A) Ovaries stained with H&E in the blank control group; (B) Magnified view of a localized area of ​​the ovary stained with H&E in the blank control group; (C) Ovaries stained with H&E in the group supplemented with 2 g / L fermented paste; (D) Magnified view of a localized area of ​​the ovary stained with H&E in the group supplemented with 2 g / L fermented paste. Detailed Implementation

[0025] The technical solution of the present invention will be described in more detail below with reference to several preferred embodiments and accompanying drawings. These embodiments are specific implementations based on the technical solution of the present invention, and provide detailed operating steps and implementation processes. However, the scope of protection of the present invention is not limited to these embodiments; any changes and equivalent substitutions within the scope of the inventive concept should be considered as protected content of the present invention.

[0026] In the following embodiments, Astragalus membranaceus was purchased from Bozhou Huizhicao Biotechnology Co., Ltd., China. Scutellaria baicalensis was purchased from Bozhou Lanhetang Health Industry Co., Ltd., China. Angelica dahurica was purchased from Bozhou Qiaocheng District Baihuayuan Food Store, China. Atractylodes macrocephala was purchased from Bozhou Guimeikang Traditional Chinese Medicine Sales Co., Ltd., China. Lonicera japonica was purchased from Bozhou Tianzhiyuan Biotechnology Co., Ltd., China. Epimedium was purchased from Bozhou Yumeng Pharmaceutical Co., Ltd., China. Honey (multifloral honey) was purchased from Mengcheng County Honggang Bee Breeding Co., Ltd., China. Glycerin was purchased from Zhengzhou Chaoxin Chemical Products Co., Ltd., China, CAS: 147-85-96. Saccharomyces cerevisiae was purchased from Jinan Jinyuyuan Biotechnology Co., Ltd., China, item number: JYY-001. Lactobacillus plantarum was purchased from Weikai Haisi (Shandong) Biotechnology Co., Ltd., China, item number: WKHS-001. Bacillus subtilis was purchased from Vikasun (Shandong) Biotechnology Co., Ltd., China, catalog number: WKHS-052. Rutin was purchased from Shanghai Maclean's Biochemical Technology Co., Ltd., China, catalog number: R817259. Sodium nitrite (NaNO2) was purchased from Shanghai Maclean's Biochemical Technology Co., Ltd., China, catalog number: S818033. Aluminum chloride (AlCl3) was purchased from Shanghai Maclean's Biochemical Technology Co., Ltd., China, catalog number: A800394. Sodium hydroxide (NaOH) was purchased from Shanghai Maclean's Biochemical Technology Co., Ltd., China, catalog number: S817971. Methanol was purchased from Shanghai Maclean's Biochemical Technology Co., Ltd., China, catalog number: M742213.

[0027] Unless otherwise specified, the experimental methods described in the following examples are all conventional techniques in the art, and are generally performed according to the techniques or conditions described in relevant literature, or with reference to the product instructions. Unless otherwise specified, the materials and reagents used can be obtained through conventional commercial channels. Example 1

[0028] (1) Raw material preparation and mixing: Weigh 20 parts of Astragalus membranaceus, 30 parts of Scutellaria baicalensis, 15 parts of Angelica dahurica, 10 parts of Atractylodes macrocephala, 15 parts of Lonicera japonica and 10 parts of Epimedium according to the weight ratio, crush each raw material, mix them evenly, and obtain the Chinese medicine mixed powder.

[0029] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:2.

[0030] (3) Addition of compound fermentation agent activation liquid: Add the compound fermentation agent activation liquid obtained in (2) to the Chinese herbal medicine mixed powder obtained in (1). The amount of activation liquid added is 4% of the total mass of the Chinese herbal medicine mixed powder, and the amount of compound fermentation agent is 2‰ of the total mass of the Chinese herbal medicine mixed powder.

[0031] (4) Moisture preparation: Add purified water to (3) in a ratio of 1:1 with the total mass of the mixed Chinese medicine powder, and stir evenly.

[0032] (5) Fermentation culture: Place the mixture in (4) at 37°C and culture for 3 days.

[0033] (6) Post-processing: The fermentation product obtained in (5) is dried at 48°C for 24 hours. Then, 3 parts honey and 3 parts glycerin are added and stirred thoroughly to obtain the fermented Chinese medicine paste. Example 2

[0034] (1) Raw material preparation and mixing: Weigh out 25 parts of Astragalus membranaceus, 25 parts of Scutellaria baicalensis, 20 parts of Angelica dahurica, 15 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica and 5 parts of Epimedium according to the weight ratio. Crush each raw material, mix them evenly, and prepare a mixed powder of Chinese medicine.

[0035] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:1.

[0036] (3) Addition of activation liquid: Add the compound fermentation agent activation liquid obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation liquid added is 16% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 8‰ of the total mass of the mixed Chinese medicine powder.

[0037] (4) Moisture preparation: Add purified water to (3) at a ratio of 1:1 to the total mass of the Chinese medicine, and stir well.

[0038] (5) Fermentation culture: Place the mixture in (4) at 42℃ and culture for 2 days.

[0039] (6) Post-processing: The fermentation product obtained in (5) is dried at 48°C for 24 hours. Then, 3 parts honey and 3 parts glycerin are added and stirred thoroughly to obtain the fermented Chinese medicine paste. Example 3

[0040] (1) Raw material preparation and mixing: Weigh out 20 parts of Astragalus membranaceus, 30 parts of Scutellaria baicalensis, 15 parts of Angelica dahurica, 20 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica and 5 parts of Epimedium according to the weight ratio. Crush each raw material, mix them evenly, and prepare a mixed powder of Chinese medicine.

[0041] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 2:1:1.

[0042] (3) Addition of activation liquid: Add the compound fermentation agent activation liquid obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation liquid added is 8% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0043] (4) Moisture preparation: Add purified water to (3) at a ratio of 1:1 to the total mass of the Chinese medicine, and stir well.

[0044] (5) Fermentation culture: Place the mixture in (4) at 42℃ and culture for 1 day.

[0045] (6) Post-processing: The fermentation product obtained in (5) is dried at 48°C for 24 hours. Then, 3 parts honey and 3 parts glycerin are added and stirred thoroughly to obtain the fermented Chinese medicine paste. Experimental Example 1 (Fermentation Time Optimization)

[0046] The fermentation time of traditional Chinese medicine fermentation broth was optimized, and the specific optimization steps are as follows: (1) Raw material preparation and mixing: Weigh out 25 parts of Astragalus membranaceus, 20 parts of Scutellaria baicalensis, 20 parts of Angelica dahurica, 20 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica and 5 parts of Epimedium according to the weight ratio. Crush each raw material, mix them evenly, and prepare a mixed powder of Chinese medicine.

[0047] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:1.

[0048] (3) Addition of activation solution: Add the compound fermentation agent activation solution obtained in step (2) to the mixed Chinese medicine powder obtained in step (1). The amount of activation solution added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0049] (4) Water preparation: Add 100 parts of purified water to step (3) and stir well.

[0050] (5) Fermentation culture: The mixture in step (4) was cultured at 37°C for 1 day, 2 days, 3 days and 4 days respectively, and the total flavonoid content after fermentation was used as the evaluation index to determine the optimal fermentation time.

[0051] (6) The total flavonoid content of different Chinese herbal fermentation broths in step (5) was detected using the following method: ① Accurately weigh 80 mg of rutin standard, dissolve it in anhydrous methanol and dilute to 10 mL to obtain a stock solution of 8 mg / mL; then, serially dilute the stock solution with ultrapure water to prepare a series of standard solutions of 4 mg / mL, 2 mg / mL, 1 mg / mL, 500 μg / mL, 250 μg / mL, 125 μg / mL, 62.5 μg / mL, 31.25 μg / mL, 15.625 μg / mL, 7.8125 μg / mL and 0 μg / mL.

[0052] ② Accurately weigh 5 g of NaNO2 and dissolve it completely in 95 mL of ultrapure water to obtain a 5% NaNO2 solution.

[0053] ③ Accurately weigh 10g of AlCl3 and dissolve it completely in 90 mL of ultrapure water to obtain a 10% AlCl3 solution.

[0054] ④ Accurately weigh 40 g of NaOH, dissolve it in ultrapure water and dilute to 1 L to prepare a 1 M NaOH solution.

[0055] ⑤ Take 0.5 mL of rutin standard solutions of different concentrations and place them in colorimetric tubes.

[0056] ⑥ Add 0.15 mL of 5% NaNO2 solution and let stand for 6 min.

[0057] ⑦ Then add 0.15 mL of 10% AlCl3 solution and let stand for 6 min.

[0058] ⑧ Add 1.0 mL of 1 M NaOH solution, mix thoroughly, and measure its absorbance at 510 nm.

[0059] ⑨ A standard curve was plotted with the concentration (y) of the rutin standard solution as the ordinate and the corresponding absorbance (x) as the abscissa, yielding the standard curve equation y = 3338x - 198.57, R0. 2 =0.9993. The results indicate that rutin showed good linearity in the range of 8 mg / mL to 7.8125 μg / mL.

[0060] ⑩ Dilute the fermentation liquid of different Chinese medicines in (5) by 20 times with ultrapure water, take the diluted sample, process it according to the process of ⑤~⑨, and measure its absorbance.

[0061] (7) The total flavonoid concentrations at different fermentation times in Example 1 were as follows: 455.40±135.16 μg / mL (after 1 day of fermentation), 4806.93±111.13 μg / mL (after 2 days of fermentation), 5265.90±259.99 μg / mL (after 3 days of fermentation) and 5161.59±186.54 μg / mL (after 4 days of fermentation). Based on comprehensive comparison, 3 days of fermentation was selected as the optimal fermentation time. Experimental Example 2 (Fermentation Temperature Optimization)

[0062] The fermentation temperature of traditional Chinese medicine fermentation broth was optimized, and the specific optimization steps are as follows: (1) Raw material preparation and mixing: Weigh 25 parts of Astragalus membranaceus, 20 parts of Scutellaria baicalensis, 20 parts of Angelica dahurica, 20 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica, and 5 parts of Epimedium according to the weight ratio of the formula described in this invention. Crush each raw material, mix them evenly, and obtain a mixed powder of traditional Chinese medicine.

[0063] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:1.

[0064] (3) Addition of activation solution: Add the compound fermentation agent activation solution obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation solution added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0065] (4) Water preparation: Add 100 parts of purified water to (3) and stir well.

[0066] (5) Fermentation culture: The mixture in (4) was placed at 27℃, 32℃, 37℃ and 42℃ and cultured for 3 days respectively. The total flavonoid content after fermentation was used as the evaluation index to determine the optimal fermentation temperature.

[0067] (6) Perform the operation as described in Example 1 (6).

[0068] (7) The total flavonoid concentrations at different fermentation temperatures in Example 2 were measured as follows: 3087.86±158.04 μg / mL (27℃), 3605.25±113.61 μg / mL (32℃), 5787.46±76.33 μg / mL (37℃) and 2770.75±68.81 μg / mL (42℃). Based on comprehensive comparison, 37℃ is the optimal fermentation temperature. Experimental Example 3 (Optimization of Fermentation Bacterial Dosage)

[0069] The specific optimization steps for the dosage of fermentation bacteria in traditional Chinese medicine fermentation broth are as follows: (1) Raw material preparation and mixing: Weigh 25 parts of Astragalus membranaceus, 20 parts of Scutellaria baicalensis, 20 parts of Angelica dahurica, 20 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica, and 5 parts of Epimedium according to the weight ratio of the formula described in this invention. Crush each raw material, mix them evenly, and obtain a mixed powder of traditional Chinese medicine.

[0070] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:1.

[0071] (3) Addition of activation liquid: Add the compound fermentation agent activation liquid obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation liquid added is 2%, 4%, 8% and 16% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 1‰, 2‰, 4‰ and 8‰ of the total mass of the mixed Chinese medicine powder.

[0072] (4) Water preparation: Add 100 parts of purified water to (3) and stir well.

[0073] (5) Fermentation culture: The mixture in (4) was placed at 37°C and cultured for 3 days. The total flavonoid content after fermentation was used as the evaluation index to determine the optimal fermentation temperature.

[0074] (6) Perform the operation as described in Example 1 (6).

[0075] (7) The total flavonoid concentrations of different fermentation bacteria dosages in Example 3 were tested as follows: 4715.13±173.98 μg / mL (1‰), 6037.81±230.61 μg / mL (2‰), 6046.16±368.28 μg / mL (4‰) and 6087.88±312.95 μg / mL (8‰); after comprehensive comparison, 2‰ was selected as the optimal fermentation bacteria dosage. Experiment 4 (Optimization of Fermentation Strains Ratio)

[0076] The specific optimization steps for the proportion of fermentation strains in the fermentation broth of traditional Chinese medicine are as follows: (1) Raw material preparation and mixing: Weigh 25 parts of Astragalus membranaceus, 20 parts of Scutellaria baicalensis, 20 parts of Angelica dahurica, 20 parts of Atractylodes macrocephala, 10 parts of Lonicera japonica, and 5 parts of Epimedium according to the weight ratio of the formula described in this invention. Crush each raw material, mix them evenly, and obtain a mixed powder of traditional Chinese medicine.

[0077] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in mass ratios of 1:1:1, 1:2:1, 1:1:2 and 2:1:1 respectively.

[0078] (3) Addition of activation solution: Add the compound fermentation agent activation solution obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation solution added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0079] (4) Water preparation: Add 100 parts of purified water to (3) and stir well.

[0080] (5) Fermentation culture: The mixture in (4) was placed at 37°C and cultured for 3 days. The total flavonoid content after fermentation was used as the evaluation index to determine the optimal fermentation temperature.

[0081] (6) Perform the operation as described in Example 1 (6).

[0082] (7) The total flavonoid concentrations of different fermentation strain ratios in Example 4 were tested as follows: 6029.47±261.91 μg / mL (1:1:1), 5904.29±224.49 μg / mL (1:2:1), 6914.04±107.63 μg / mL (1:1:2) and 6413.34±137.21 μg / mL (2:1:1); after comprehensive comparison, 1:1:2 is the optimal fermentation strain ratio. Experimental Example 5 (Optimization of the quality ratio of traditional Chinese medicine compound prescriptions)

[0083] The specific optimization steps for the proportion of fermentation strains in the fermentation broth of traditional Chinese medicine are as follows: (1) Raw material preparation and mixing: Weigh each raw material according to the following weight ratio, crush them, mix them evenly, and obtain Chinese medicine mixed powder.

[0084] Ratio ①: Astragalus membranaceus 25 parts, Scutellaria baicalensis 20 parts, Angelica dahurica 20 parts, Atractylodes macrocephala 20 parts, Lonicera japonica 10 parts and Epimedium 5 parts; Ratio ②: Astragalus membranaceus 20 parts, Scutellaria baicalensis 25 parts, Angelica dahurica 20 parts, Atractylodes macrocephala 20 parts, Lonicera japonica 10 parts and Epimedium 5 parts; Ratio ③: Astragalus membranaceus 15 parts, Scutellaria baicalensis 25 parts, Angelica dahurica 25 parts, Atractylodes macrocephala 20 parts, Lonicera japonica 10 parts and Epimedium 5 parts; Ratio ④: 10 parts Astragalus membranaceus, 25 parts Scutellaria baicalensis, 25 parts Angelica dahurica, 25 parts Atractylodes macrocephala, 10 parts Lonicera japonica, and 5 parts Epimedium; Ratio ⑤: Astragalus membranaceus 20 parts, Scutellaria baicalensis 30 parts, Angelica dahurica 15 parts, Atractylodes macrocephala 10 parts, Lonicera japonica 15 parts and Epimedium 10 parts; Ratio ⑥: Astragalus membranaceus 25 parts, Scutellaria baicalensis 25 parts, Angelica dahurica 15 parts, Atractylodes macrocephala 15 parts, Lonicera japonica 10 parts and Epimedium 10 parts; (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:2.

[0085] (3) Addition of activation solution: Add the compound fermentation agent activation solution obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation solution added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0086] (4) Water preparation: Add 100 parts of purified water to (3) and stir well.

[0087] (5) Fermentation culture: The mixture in (4) was placed at 37°C and cultured for 3 days. The total flavonoid content after fermentation was used as the evaluation index to determine the optimal mass ratio of the traditional Chinese medicine compound.

[0088] (6) Perform the operation as described in Example 1 (6).

[0089] (7) The total flavonoid concentrations of different Chinese herbal compound mass ratios in Example 5 were tested as follows: 6826.42±216.97 μg / mL (ratio ①), 7164.39±280.73 μg / mL (ratio ②), 6964.11±183.53 μg / mL (ratio ③), 6847.28±118.31 μg / mL (ratio ④), 8107.37±281.72 μg / mL (ratio ⑤) and 7648.40±123.68 μg / mL (ratio ⑥). Based on comprehensive comparison, ratio ⑤ is the optimal Chinese herbal compound mass ratio. Experimental Example 6 (Animal experimental production performance, immune performance, and pathological tissue sections)

[0090] The preparation steps of fermented Chinese herbal medicine paste are as follows: (1) Raw material preparation and mixing: Weigh out 20 parts of Astragalus membranaceus, 30 parts of Scutellaria baicalensis, 15 parts of Angelica dahurica, 10 parts of Atractylodes macrocephala, 15 parts of Lonicera japonica and 10 parts of Epimedium according to the weight ratio. Crush each raw material, mix them evenly, and prepare a mixed powder of Chinese medicine.

[0091] (2) Activation of compound fermentation agent: Mix compound fermentation agent, brown sugar and water in a mass ratio of 1:1:18 and stir thoroughly until uniform to obtain compound fermentation agent activation solution. The compound fermentation agent is composed of brewer's yeast, lactobacillus plantarum and Bacillus subtilis in a mass ratio of 1:1:2.

[0092] (3) Addition of activation solution: Add the compound fermentation agent activation solution obtained in (2) to the mixed Chinese medicine powder obtained in (1). The amount of activation solution added is 4% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 2‰ of the total mass of the mixed Chinese medicine powder.

[0093] (4) Water preparation: Add 100 parts of purified water to (3) and stir well.

[0094] (5) Fermentation culture: Place the mixture in (4) at 37°C and culture for 3 days.

[0095] (6) Post-processing: The fermentation product obtained in (5) is dried at 48°C for 24 hours. Then, 3 parts honey and 3 parts glycerin are added and stirred thoroughly to obtain the fermented Chinese medicine paste.

[0096] To systematically evaluate the regulatory effects of fermented traditional Chinese medicine (TCM) extract on the laying performance and immune function of laying hens, this experiment designed two treatment groups: a blank control group (normal diet) and a TCM fermented extract treatment group (normal diet + 2 g / L TCM fermented extract). The study subjects were 42,600 and 43,200 one-day-old Nongda No. 3 laying hens, respectively assigned to the control group and the experimental group. The experiment lasted for 260 days, during which the TCM fermented extract was provided to the laying hens in their drinking water.

[0097] As shown in Table 1, the addition of 2 g / L fermented extract to the diet significantly promoted the laying process in laying hens. Specifically, at 140 days of age, compared with the control group, the egg production rate of the fermented extract group reached 69.5%, and its feed conversion ratio and feed-to-egg ratio decreased by 31.34% and 27.70%, respectively, indicating that the additive significantly improved feed utilization efficiency. At 170 days of age, the egg production rate of the fermented extract group further increased to 95.1%, significantly higher than that of the control group. The data from the entire rearing to early laying period show that the fermented extract group exhibited the most significant feed conversion advantage during the 140–230 day age period, indicating that the traditional Chinese medicine compound fermented extract can play a significant role in promoting laying, improving feed utilization, and optimizing early laying performance.

[0098] As shown in Tables 2 and 3, adding 2 g / L of fermented extract can effectively reduce the dispersion of antibody titers against various avian influenza viruses and Newcastle disease viruses in laying hens, making the immune levels among the flock more uniform and further improving the overall health of the flock. These results suggest that fermented extract not only improves production performance but also has good immunomodulatory and flock stability-improving effects, providing important technical support for large-scale laying hen farming.

[0099] The independently developed traditional Chinese medicine compound can generally improve antibody levels. The antibody levels of the group with added fermented traditional Chinese medicine were significantly better than those of the blank control group and the group with added traditional Chinese medicine, indicating that the screened traditional Chinese medicine, especially fermented traditional Chinese medicine, can effectively enhance the specific immune response induced by the vaccine without the addition of anti-stress drugs.

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[0102]

[0103] Depend on Figure 1 As shown, the liver tissues of two groups of laying hens were pathologically observed using H&E staining. The liver lobule structure of the blank control group was relatively intact, but mild cellular degeneration and edema were observed in some areas. A small number of lymphocytes and monocytes infiltrated the interlobular spaces, indicating a possible mild immune response in the liver. Although the hepatocytes were arranged relatively regularly, a small number of hepatocytes showed mild vacuolation or swelling. In contrast, liver sections from the group supplemented with 2 g / L of fermented traditional Chinese medicine extract showed normal hepatocyte arrangement, intact liver lobule structure, and no obvious abnormalities in the intercellular matrix. This group's liver tissue did not show cell necrosis or severe inflammatory reactions, indicating that the fermented traditional Chinese medicine extract did not cause significant pathological damage. These results indicate that the liver sections from the group supplemented with fermented traditional Chinese medicine extract showed a healthier liver tissue structure compared to the blank control group, with no obvious pathological changes. This suggests that during the 260-day experimental period, the fermented traditional Chinese medicine extract had no significant adverse effects on the liver of laying hens and may even help maintain normal liver function.

[0104] Depend on Figure 2As shown, the pathological observation of the ovaries of laying hens in the blank control group and the group treated with traditional Chinese medicine fermentation paste was performed using H&E staining. Ovarian sections from the blank control group showed significant pathological damage, mainly manifested as abnormal follicular development and damage to the ovarian tissue structure. Multiple follicles showed varying degrees of atrophy and cavitation, indicating that normal follicular development was inhibited. Simultaneously, significant fatty degeneration was observed within the ovary, with a large accumulation of adipocytes in some areas, accompanied by fibrosis and hyperplasia in some regions, leading to a loose ovarian stroma structure. Vasodilation and erythrocyte infiltration were evident, with local congestion, suggesting that the ovarian tissue may be affected by insufficient blood supply and chronic inflammation, resulting in overall ovarian function inhibition, possibly related to metabolic disorders or long-term inflammatory responses. In contrast, ovarian sections from the group treated with traditional Chinese medicine fermentation paste showed significant pathological improvement. Although some follicles still showed morphological abnormalities or developmental arrest, the overall follicular development arrest was significantly reduced, the follicular structure remained relatively intact, and the degree of cavitation was also reduced. Fatty degeneration was significantly reduced compared to the control group, the area of ​​adipocyte infiltration decreased, and the ovarian tissue structure became more compact. The degree of fibrosis was also reduced, the structure of the ovarian stroma was better maintained, and no significant fibrosis was observed. Vasodilation and erythrocyte infiltration were significantly improved, local inflammatory response was significantly reduced, and the integrity of the blood vessel walls was restored. These results indicate that, during the experimental period, the addition of 2 g / L of fermented traditional Chinese medicine extract significantly improved the pathological state of the ovaries of laying hens. Compared with the blank control group, the ovarian tissue of the treated group showed a healthier structure and function.

[0105] In summary, this invention utilizes natural Chinese medicinal herbs through fermentation to produce a fermented herbal paste, which is then dissolved in drinking water for feeding. Adding 2 g / L of this fermented paste to the diet of laying hens significantly improves their production performance and immune function. Studies have shown that adding the fermented herbal paste not only promotes the onset of egg production and increases egg production rate, but also significantly improves feed utilization efficiency, exhibiting the most significant feed conversion advantage, especially in the 140-230 day age range. Simultaneously, the fermented herbal paste enhances antibody levels by regulating the immune system of laying hens, improving their immune response to avian influenza and Newcastle disease viruses, and thus improving the overall health of the flock. Pathological analysis shows that the liver and ovarian tissues of the group receiving the fermented herbal paste exhibited healthier structure and function, with no obvious pathological damage, and may help maintain normal liver and ovarian function.

[0106] The fermented herbal paste of this invention can be widely used in laying hen feed. It can be fed through a drinking water system, which is quick and convenient. It can effectively improve the production performance and immunity of laying hens, optimize breeding efficiency, and has important technical and economic value.

Claims

1. A fermented traditional Chinese medicine extract for improving the production performance and immune function of laying hens, characterized in that, It is made from the following components in parts by weight: Astragalus membranaceus 5-30 parts, Scutellaria baicalensis 5-30 parts, Angelica dahurica 5-20 parts, Atractylodes macrocephala 5-20 parts, Lonicera japonica 5-20 parts, and Epimedium 5-10 parts.

2. The fermented herbal paste according to claim 1, characterized in that... It is made from the following components in parts by weight: Astragalus membranaceus 20-25 parts, Scutellaria baicalensis 20-30 parts, Angelica dahurica 15-20 parts, Atractylodes macrocephala 10-20 parts, Lonicera japonica 10-15 parts, and Epimedium 5-10 parts.

3. The fermented herbal paste according to claim 1, characterized in that... It is made from the following components in parts by weight: 20 parts Astragalus membranaceus, 30 parts Scutellaria baicalensis, 15 parts Angelica dahurica, 10 parts Atractylodes macrocephala, 15 parts Lonicera japonica, and 10 parts Epimedium.

4. A method for preparing a fermented herbal extract according to any one of claims 1 to 3, characterized in that... Includes the following steps: (1) Mix the above-mentioned dried Chinese medicine powders in a certain proportion to obtain a mixed Chinese medicine powder; (2) Add a compound fermentation agent activation solution composed of compound fermentation agent, brown sugar and water to the mixed powder of Chinese medicine; the mass ratio of the compound fermentation agent, brown sugar and water is 1:1:

18. (3) Add water to the mixture obtained in step (2) and ferment it. The fermentation temperature is 27~42℃ and the fermentation time is 1~4 days. Dry it into a paste.

5. The method for preparing fermented herbal extract according to claim 4, characterized in that, The ratio of water added in step (3) to the total mass of the Chinese medicine is 1:

1.

6. The method for preparing fermented herbal extract according to claim 4, characterized in that, The amount of compound fermentation agent activation liquid added in step (2) is 2-16% of the total mass of the mixed Chinese medicine powder, and the amount of compound fermentation agent is 1-8‰ of the total mass of the mixed Chinese medicine powder.

7. The method for preparing fermented herbal extract according to claim 4, characterized in that, The compound fermentation agent is a combination of Saccharomyces cerevisiae, Lactobacillus plantarum and Bacillus subtilis in a ratio of 1:1:1, 1:2:1, 1:1:2 or 2:1:

1.

8. The method for preparing fermented herbal extract according to claim 4, characterized in that, Add 1-5 parts honey and 1-5 parts glycerin to the paste obtained in step (3) and mix thoroughly.

9. The use of the fermented herbal extract according to any one of claims 1 to 3 in the preparation of feed additives that improve the laying hen's production performance and immune performance.

10. The application according to claim 9, wherein the improvement of laying hen production performance is achieved by the fermented Chinese medicine paste promoting the laying process and improving the pathological state of the ovary and liver of the laying hen.

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