Fermented traditional Chinese medicine compound, preparation method and application thereof

Abstract

This application belongs to the technical field of traditional Chinese medicine compound formulas for poultry, and discloses a fermented traditional Chinese medicine compound, its preparation method, and its application. The fermented traditional Chinese medicine compound is obtained by fermenting a traditional Chinese medicine compound with Aspergillus niger. The traditional Chinese medicine compound comprises the following components by weight: 70-90 parts Coptis chinensis, 90-110 parts Scutellaria baicalensis, 110-130 parts Lonicera japonica, 90-110 parts Forsythia suspensa, 110-130 parts Pulsatilla chinensis, 90-110 parts Andrographis paniculata, 110-130 parts Taraxacum mongolicum, 70-90 parts Atractylodes lancea, 90-110 parts Atractylodes macrocephala, 90-110 parts Poria cocos, and 30-50 parts Glycyrrhiza uralensis. This application provides a fermented traditional Chinese medicine compound that, through fermentation with Aspergillus niger using a specific combination of traditional Chinese medicines, significantly improves the prevention and treatment efficacy of the traditional Chinese medicine combination against bacterial diseases in chickens.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine compound formulas for poultry, specifically to a fermented traditional Chinese medicine compound formula, its preparation method, and its application. Background Technology

[0002] Bacterial diseases are one of the major risks facing the poultry industry. Their high incidence, mortality, and adaptability to the environment mean that poultry farms must invest enormous effort and resources in hygiene management for prevention and control. Among them, pullorum disease, fowl cholera (fowl pasteurellosis), and colibacillosis are the most common and serious bacterial diseases in poultry production, especially in chicks and growing chickens, where they are prone to causing diarrhea, septicemia, and high mortality rates, seriously affecting the profitability of poultry farming.

[0003] Currently, the prevention and control of the above-mentioned diseases still mainly rely on antibiotics. However, long-term or irregular use can easily lead to increased bacterial resistance, increased risk of drug residues, and imbalance of the intestinal microecology, which can hardly meet the development needs of antibiotic reduction, antibiotic alternatives and green farming.

[0004] Prior art 1: Chinese patent application 201710669519.2 discloses a traditional Chinese medicine composition for preventing and treating bacterial diseases of the digestive tract in poultry, its preparation method and application, which is mainly made from the following raw materials: 20-80 parts of Bidens pilosa, 20-80 parts of garlic, and 5-20 parts of vitamin C; Prior art 1 uses Bidens pilosa, garlic and vitamin C to form a traditional Chinese medicine composition to prevent bacterial diseases, which can effectively avoid the effects of antibiotics.

[0005] However, traditional Chinese medicine powders or decoctions have shortcomings such as insufficient release of effective ingredients and low bioavailability, which limit their practical application effects. Summary of the Invention

[0006] One of the purposes of this application is to provide a fermented traditional Chinese medicine compound that can fully utilize the active ingredients of the traditional Chinese medicine compound and effectively improve its application effect in the prevention and treatment of bacterial diseases in chickens.

[0007] Another objective of this application is to provide a fermented traditional Chinese medicine compound that produces a fermented traditional Chinese medicine compound with a high rate of prevention and treatment of bacterial diseases in chickens.

[0008] Meanwhile, this application also provides a fermented traditional Chinese medicine compound that can be applied to the preparation of feed additives or drugs for poultry bacterial diseases, thereby bringing the efficacy of feed additives and drugs in preventing and treating poultry bacterial diseases.

[0009] To achieve the above objectives, this application provides a fermented traditional Chinese medicine compound, obtained by fermentation of a traditional Chinese medicine compound with Aspergillus niger. The traditional Chinese medicine compound comprises the following components by weight: 70-90 parts Coptis chinensis, 90-110 parts Scutellaria baicalensis, 110-130 parts Lonicera japonica, 90-110 parts Forsythia suspensa, 110-130 parts Pulsatilla chinensis, 90-110 parts Andrographis paniculata, 110-130 parts Taraxacum mongolicum, 70-90 parts Atractylodes lancea, 90-110 parts Atractylodes macrocephala, 90-110 parts Poria cocos, and 30-50 parts Glycyrrhiza uralensis.

[0010] This application also provides a method for preparing the above-mentioned fermented traditional Chinese medicine compound, comprising the following steps:

[0011] Step 1: Prepare the traditional Chinese medicine compound into a matrix;

[0012] Step 2: Inoculate the substrate with Aspergillus niger for fermentation to obtain the fermented traditional Chinese medicine compound.

[0013] Furthermore, the specific operation of step 1 is as follows: Coptis chinensis, Scutellaria baicalensis, Lonicera japonica, Forsythia suspensa, Pulsatilla chinensis, Andrographis paniculata, Taraxacum mongolicum, Atractylodes lancea, Atractylodes macrocephala, Poria cocos and Glycyrrhiza uralensis are pulverized into powder, and then mixed with 8 times the mass of the powder to prepare a moist matrix. The moist matrix is ​​then sterilized under high pressure at 121°C for 25 minutes and cooled to 32°C to obtain the matrix.

[0014] Furthermore, the specific operation of step 2 is as follows: Inoculate the substrate with spores at a concentration of 1×10⁻⁶ at a wet weight ratio of 1% (v / w). 7 CFU / ml is a suspension of Aspergillus niger spores; then solid-state fermentation is carried out at 30-33℃ for 72 hours to obtain fermented traditional Chinese medicine compound.

[0015] Preferably, the specific process of the solid-state fermentation is as follows: the relative humidity of the environment is maintained at 60-70%, and the initial pH is controlled at 5.0-6.0; the material is turned over once every 24 hours during the fermentation period.

[0016] In a further preferred embodiment, the preparation method of the fermented traditional Chinese medicine compound also includes step 3: drying the fermented traditional Chinese medicine compound at 60°C until the moisture content is less than 10%, then pulverizing and passing it through a 40-mesh sieve.

[0017] This application also provides the application of drugs for the prevention and treatment of bacterial diseases in poultry prepared using the above-mentioned fermented traditional Chinese medicine compound.

[0018] This application also provides the application of feed additives prepared using the above-mentioned fermented traditional Chinese medicine compound for the prevention and treatment of bacterial diseases in poultry.

[0019] Beneficial effects

[0020] Compared with the prior art, this application provides a fermented traditional Chinese medicine compound, which uses a specific combination of traditional Chinese medicines fermented with Aspergillus niger, which greatly improves the prevention and treatment effect of the traditional Chinese medicine combination on bacterial diseases in chickens. Detailed Implementation

[0021] The present application will be further described below with reference to embodiments, but this does not constitute any limitation on the present application. Any limited modifications made within the scope of the claims of the present application shall still be within the scope of the claims of the present application.

[0022] To illustrate the technical content of this application in detail, the following description is provided in conjunction with the embodiments.

[0023] Example 1

[0024] A fermented traditional Chinese medicine compound is prepared using the following steps:

[0025] Step 1: Weigh out 80g of Coptis chinensis, 100g of Scutellaria baicalensis, 120g of Lonicera japonica, 100g of Forsythia suspensa, 120g of Pulsatilla chinensis, 100g of Andrographis paniculata, 120g of Taraxacum mongolicum, 80g of Atractylodes lancea, 100g of Atractylodes macrocephala, 100g of Poria cocos, and 40g of Glycyrrhiza uralensis. Grind and mix them until they can all pass through a 30-mesh sieve to obtain powder.

[0026] Step 2: Add 8 times the weight of the powder to deionized water to prepare a moist matrix. Autoclave the moist matrix at 121°C for 25 min and then cool it to 32°C to obtain the matrix.

[0027] Step 3: Inoculate the substrate with a spore concentration of 1×10⁻⁶ at a ratio of 1% (v / w) based on spore concentration. 7 The CFU / ml of Aspergillus niger spore suspension was then subjected to solid-state fermentation at 30-33℃ for 72 hours to obtain the fermented traditional Chinese medicine compound. The relative humidity of the solid-state fermentation environment was maintained at 60-70%, and the initial pH was controlled at 5.0-6.0. The material was turned over once every 24 hours during the fermentation period.

[0028] Step 4: After solid-state fermentation in Step 3, the fermented Chinese herbal medicine compound is dried at 60°C until the moisture content is less than 10%, then pulverized and passed through a 40-mesh sieve to obtain powdered fermented Chinese herbal medicine compound.

[0029] Comparative Example 1

[0030] A traditional Chinese medicine compound is prepared using the following steps:

[0031] Step 1: Weigh out 80g of Coptis chinensis, 100g of Scutellaria baicalensis, 120g of Lonicera japonica, 100g of Forsythia suspensa, 120g of Pulsatilla chinensis, 100g of Andrographis paniculata, 120g of Taraxacum mongolicum, 80g of Atractylodes lancea, 100g of Atractylodes macrocephala, 100g of Poria cocos, and 40g of Glycyrrhiza uralensis. Grind and mix them until they can all pass through a 30-mesh sieve to obtain powder.

[0032] Efficacy Experiment

[0033] 1. Animal Experiment Design

[0034] The experiment was conducted in chicks (1–14 days old) and pullets (15–42 days old). Pullorum disease and E. coli infection models were established in each stage, and a healthy control group, an infected model group, a florfenicol control group, a comparative example group, and an example group were set up. In the chick stage, each group had 3 replicates with 30 birds per replicate (90 birds per group); in the pullet stage, each group had 3 replicates with 20 birds per replicate (60 birds per group). Chickens were randomly assigned to each group and housed in independent pens.

[0035] 2. Administration method and dosage

[0036] Both Example 1 and Comparative Example 1 were administered as feed additives, mixed thoroughly with the basal diet at a dosage of 1.5 kg / ton of complete feed. Both traditional Chinese medicine preparations were pre-fed 3 days before challenge and continued to be used until day 14 after challenge. The florfenicol control group received the medication via drinking water at the recommended dosage as per the product instructions, converted to an effective ingredient concentration of 20 mg / kg body weight / day, for 5 consecutive days (days 0-4 after challenge). During the remaining time, the control group was fed the basal diet without the use of other antibiotics.

[0037] 3. Observation indicators

[0038] During the experiment, the mental state, feed intake, and fecal characteristics of the chickens were observed daily, and the severity of diarrhea and mental state were scored. The incidence, morbidity, mortality, and duration of the disease were also recorded. Average daily gain (ADG) and feed conversion ratio (FCR) were measured in chicks and growers. Furthermore, feces and liver and spleen tissue from some chickens were collected on days 3 and 7 post-infection to determine the pathogen load, evaluating the inhibitory effects of different treatments on pathogen colonization and systemic dissemination.

[0039] 4. Results

[0040] 4.1 The preventive and therapeutic effects of fowl typhoid fever

[0041] 4.1.1 The effect of preventing and treating fowl cholera in chicks

[0042] Table 1 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group for the fowl typhoid model: Diarrhea incidence = number of fowl with diarrhea ÷ number of experimental fowl × 100%; Mortality rate = number of dead fowl ÷ number of experimental fowl × 100%;

[0043] Table 1. Comparison of clinical outcomes and growth performance among different treatment groups in a fowl typhoid model.

[0044] Group Incidence of diarrhea (%) Course of illness (d) mortality rate(%) ADG (g / d) FCR Infection model group 42.5±4.8 7.8±1.3 18.0±3.1 14.2±1.6 2.15±0.12 Florfenicol group 18.6±3.9 4.9±1.0 6.8±1.9 18.9±1.5 1.86±0.10 Comparative Example 1 25.8±4.1 5.2±1.0 9.6±2.2 17.5±1.4 1.92±0.11 Example 1 Group 12.4±3.2 3.6±0.8 4.2±1.5 20.1±1.5 1.78±0.10

[0045] After establishing a pullorum disease model in chicks, the infected model group showed obvious depression, decreased feed intake and persistent diarrhea, and some chicks died within 3–7 days after the challenge. Compared with the infected model group, the florfenicol group, comparative example 1 group and example 1 group all improved clinical symptoms and reduced mortality to varying degrees.

[0046] Among them, Example 1 group showed the most significant results in reducing the incidence of diarrhea, shortening the course of the disease and improving the survival rate. Compared with Comparative Example 1 group, the incidence of diarrhea and mortality rate were significantly improved. Combined with the florfenicol group, it can be seen that although the traditional Chinese medicine compound alone has a certain preventive and therapeutic effect on chicken white diarrhea, it can only significantly improve its preventive effect after fermentation by Aspergillus niger.

[0047] 4.1.2 Pathogen detection of fowl cholera in chicks

[0048] Regarding etiological detection, feces and liver and spleen tissues were collected on the 3rd and 7th day after challenge for quantitative analysis. The results are shown in Table 2.

[0049] Table 2 Comparison of pathogen load after challenge in chicken pullorum model (log10 CFU / g)

[0050] Time point Group Fecal load (log10 CFU / g) Liver / spleen load (log10 CFU / g) d3 Infection model group 7.2±0.5 5.9±0.6 d3 Florfenicol group 5.9±0.4 4.6±0.5 d3 Comparative Example 1 6.4±0.4 5.1±0.5 d3 Example 1 Group 5.6±0.4 4.3±0.4 d7 Infection model group 6.5±0.6 5.0±0.6 d7 Florfenicol group 5.4±0.5 3.9±0.5 d7 Comparative Example 1 5.8±0.5 4.2±0.5 d7 Example 1 Group 4.9±0.5 3.4±0.4

[0051] The results in Table 2 show that the pathogen load in Example 1 group was significantly lower than that in the infection model group, Comparative Example 1 group and Florfenicol group at both time points, suggesting that it has a better effect in inhibiting pathogen colonization and systemic dissemination.

[0052] 4.1.3 Prevention and treatment effects of pullorum disease in growing chickens

[0053] Table 3 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group for the pullet white diarrhea model. The calculation methods for diarrhea incidence and mortality are the same as those for chicks.

[0054] Table 3 Comparison of prevention and control effects among different groups in the pullet white diarrhea model.

[0055] Group Incidence of diarrhea (%) Course of illness (d) mortality rate(%) ADG (g / d) FCR Infection model group 34.8±5.0 6.9±1.1 7.6±2.0 31.2±2.1 2.08±0.11 Florfenicol group 14.2±3.8 4.0±0.8 2.3±1.1 36.0±2.2 1.85±0.09 Comparative Example 1 22.6±4.2 4.8±0.9 3.9±1.4 34.1±2.0 1.93±0.10 Example 1 Group 11.3±3.1 3.5±0.7 1.5±0.9 37.2±2.3 1.77±0.08

[0056] After establishing a pullorum model in growing chickens, the infected model group mainly exhibited persistent diarrhea, growth retardation, and a certain mortality rate. Compared with the infected model group, the florfenicol group, comparative example 1 group, and example 1 group all significantly improved clinical symptoms and reduced mortality. Among them, example 1 group showed the best performance in terms of diarrhea incidence, duration of disease, and recovery of growth performance.

[0057] 4.1.4 Pathogen detection of pullorum disease in growing chickens

[0058] Salmonella load in feces and liver and spleen tissues of pullets were measured on days 3 and 7 after challenge. The results are shown in Table 4.

[0059] Table 4. Salmonella load in pullet model chickens after challenge (log10 CFU / g)

[0060] Time point Group Fecal load (log10 CFU / g) Liver / spleen load (log10 CFU / g) d3 Infection model group 6.9±0.5 5.5±0.6 d3 Florfenicol group 5.6±0.4 4.3±0.5 d3 Comparative Example 1 6.1±0.4 4.8±0.5 d3 Example 1 Group 5.2±0.4 3.9±0.4 d7 Infection model group 6.2±0.6 4.8±0.5 d7 Florfenicol group 5.1±0.5 3.8±0.4 d7 Comparative Example 1 5.5±0.5 4.1±0.4 d7 Example 1 Group 4.6±0.4 3.2±0.4

[0061] The results showed that, compared with the infection model group, the florfenicol group, comparative example 1 group, and example 1 group all reduced bacterial load. Among them, example 1 group had the lowest fecal and organ load at both time points, suggesting that it has a better effect in inhibiting intestinal colonization and systemic dissemination.

[0062] 4.2 The effectiveness of prevention and control of Escherichia coli infection

[0063] 4.2.1 The effect of prevention and control of Escherichia coli infection in chicks

[0064] Table 5 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group in the Escherichia coli infection model of chicks.

[0065] Table 5 Comparison of control effects among different groups in the E. coli infection model in chicks

[0066] Group Incidence rate (%) Course of illness (d) mortality rate(%) ADG (g / d) FCR Infection model group 44.2±5.6 8.1±1.4 16.7±3.0 13.8±1.5 2.22±0.13 Florfenicol group 19.5±4.2 5.0±1.0 6.4±1.8 18.5±1.6 1.88±0.11 Comparative Example 1 27.8±4.8 5.8±1.1 9.2±2.1 16.9±1.5 1.97±0.12 Example 1 Group 13.6±3.5 3.9±0.8 4.1±1.4 19.6±1.7 1.80±0.10

[0067] After establishing an E. coli infection model in chicks, the infected model group mainly exhibited depression, diarrhea, and significantly stunted growth, with some chickens developing septicemia and dying. Compared with the infected model group, all treatment groups significantly reduced morbidity and mortality. Among them, the Example 1 group showed the most significant performance in reducing morbidity, shortening the course of the disease, and improving survival rate.

[0068] 4.2.2 Pathogen detection of Escherichia coli infection in chicks

[0069] In the chick colibacillosis model, the Escherichia coli load in feces and liver and spleen tissues was measured on days 3 and 7 after challenge. The results are shown in Table 6.

[0070] Table 6. Escherichia coli load after challenge in chick E. coli infection model (log10 CFU / g)

[0071] Time point Group fecal load Liver / spleen load d3 Infection model group 7.4±0.5 6.0±0.6 d3 Florfenicol group 6.0±0.4 4.7±0.5 d3 Comparative Example 1 6.5±0.4 5.2±0.5 d3 Example 1 Group 5.7±0.4 4.4±0.4 d7 Infection model group 6.8±0.6 5.4±0.5 d7 Florfenicol group 5.6±0.5 4.1±0.4 d7 Comparative Example 1 6.0±0.5 4.5±0.4 d7 Example 1 Group 5.0±0.5 3.6±0.4

[0072] The results showed that the bacterial load in Example 1 group was significantly lower than that in the infection model group, Comparative Example 1 group and Florfenicol group at both time points.

[0073] 4.2.3 The effect of prevention and control of Escherichia coli infection in pullets

[0074] Table 7 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group in the Escherichia coli infection model of growing chickens.

[0075] Table 7 Comparison of control effects among different groups in the E. coli infection model of pullets

[0076] Group Incidence rate (%) Course of illness (d) mortality rate(%) ADG (g / d) FCR Infection model group 36.7±5.2 7.8±1.2 8.5±2.1 30.1±2.0 2.15±0.10 Florfenicol group 12.8±3.6 4.1±0.9 2.6±1.2 36.2±2.1 1.86±0.09 Comparative Example 1 21.3±4.0 5.2±0.9 4.0±1.5 34.0±2.1 1.92±0.09 Example 1 Group 10.6±3.1 3.6±0.7 1.8±1.0 36.9±2.2 1.78±0.08

[0077] After establishing an E. coli infection model in pullets, the infected model group mainly exhibited lethargy, stunted growth, and varying degrees of respiratory symptoms. Compared with the infected model group, all treatment groups significantly reduced morbidity and mortality, and improved growth performance.

[0078] Among them, Example 1 group showed the best performance in key indicators such as incidence rate, duration of disease course and average daily weight gain, with an overall effect superior to Comparative Example 1 group, and superior to florfenicol group in most endpoints.

[0079] 4.2.4 Pathogen detection of Escherichia coli infection in pullets

[0080] In the Escherichia coli infection model in growing chickens, feces and liver and spleen tissues of chickens in each group were collected on the 3rd and 7th day after challenge, and the Escherichia coli load was quantitatively detected. The results are shown in Table 8.

[0081] Table 8. Escherichia coli load (log10 CFU / g) after challenge in pullet chicken E. coli infection model

[0082] Time point Group fecal load Liver / spleen load d3 Infection model group 7.0±0.5 5.8±0.6 d3 Florfenicol group 5.7±0.4 4.5±0.5 d3 Comparative Example 1 6.2±0.4 5.0±0.5 d3 Example 1 Group 5.3±0.4 4.1±0.4 d7 Infection model group 6.4±0.6 5.1±0.5 d7 Florfenicol group 5.2±0.5 4.0±0.4 d7 Comparative Example 1 5.6±0.5 4.4±0.4 d7 Example 1 Group 4.7±0.5 3.5±0.4

[0083] The results showed that, compared with the infection model group, the florfenicol group, Comparative Example 1 group, and Example 1 group all exhibited varying degrees of viral load reduction at both time points. Among them, Example 1 group had the lowest fecal and organ viral loads at d3 and d7, which were lower than those of the florfenicol group and Comparative Example 1 group, suggesting that it has a better effect in inhibiting intestinal colonization and reducing systemic dissemination.

[0084] 4.3 Prevention and control effects of fowl cholera

[0085] 4.3.1 Prevention and control efficacy of fowl cholera in chicks

[0086] Table 9 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group in the fowl cholera model of chicks.

[0087] Table 9. Comparison of clinical outcomes among different groups in the fowl cholera model in chicks.

[0088] Group Incidence rate (%) Course of illness (d) Acute mortality rate (%) Infection model group 52.6±6.4 5.2±0.8 24.8±3.7 Florfenicol group 22.4±4.6 3.3±0.6 8.2±2.0 Comparative Example 1 31.7±5.1 3.7±0.7 11.6±2.5 Example 1 Group 16.8±3.9 2.6±0.5 5.4±1.6

[0089] The fowl cholera model in chicks is characterized by acute onset and high mortality; after challenge, the infected chickens in the model group showed depression, loss of appetite and mass mortality within a short period of time.

[0090] Compared with the infection model group, all treatment groups significantly reduced acute mortality and shortened the course of the disease, with the protective effect of Group 1 of No. 14 being the most obvious.

[0091] 4.3.2 Pathogen detection of fowl cholera in chicks

[0092] In a chick fowl cholera model, the Pasteurella multocida load in feces and liver and spleen tissues was measured on days 3 and 7 after challenge. The results are shown in Table 10.

[0093] Table 10 Pasteurella multocida load after challenge in fowl cholera model of chicks (log10 CFU / g)

[0094] Time point Group fecal load Liver / spleen load d3 Infection model group 7.1±0.6 6.2±0.6 d3 Florfenicol group 5.8±0.5 4.9±0.5 d3 Comparative Example 1 6.3±0.5 5.3±0.5 d3 Example 1 Group 5.4±0.4 4.5±0.4 d7 Infection model group 6.5±0.6 5.6±0.5 d7 Florfenicol group 5.3±0.5 4.2±0.4 d7 Comparative Example 1 5.7±0.5 4.6±0.4 d7 Example 1 Group 4.8±0.5 3.8±0.4

[0095] The results showed that the load in Example 1 group was the lowest at both time points, and lower than that in the florfenicol group and Comparative Example 1 group.

[0096] 4.3.3 Prevention and control effect of fowl cholera in pullets

[0097] Table 11 shows the prevention and control effects of the infected model group, florfenicol control group, comparative example 1 group, and example 1 group compared with the healthy control group in the fowl cholera model of growing chickens.

[0098] Table 11 Comparison of clinical outcomes among different groups in the fowl cholera model in pullets

[0099] Group Incidence rate (%) Course of illness (d) Acute mortality rate (%) Infection model group 46.1±6.0 5.0±0.9 19.3±3.1 Florfenicol group 19.6±4.1 3.2±0.7 6.9±1.9 Comparative Example 1 27.4±4.7 3.6±0.8 9.8±2.3 Example 1 Group 14.9±3.5 2.7±0.6 4.3±1.5

[0100] After establishing a fowl cholera model in pullets, the infected model group exhibited acute onset, depression, and a certain percentage of mortality.

[0101] Compared with the infection model group, all treatment groups significantly reduced mortality and shortened the course of disease, with the Example 1 group showing the best performance in controlling acute death.

[0102] 4.3.4 Pathogen detection of fowl cholera in pullets

[0103] In the fowl cholera model in growing chickens, the Pasteurella multocida load in feces and liver and spleen tissues was measured on days 3 and 7 after challenge. The results are shown in Table 12.

[0104] Table 12 Pasteurella multocida load after challenge in fowl cholera model in growing chickens (log10 CFU / g)

[0105] Time point Group fecal load Liver / spleen load d3 Infection model group 6.8±0.6 5.9±0.6 d3 Florfenicol group 5.6±0.5 4.7±0.5 d3 Comparative Example 1 6.0±0.5 5.1±0.5 d3 Example 1 Group 5.1±0.4 4.2±0.4 d7 Infection model group 6.1±0.6 5.2±0.5 d7 Florfenicol group 5.0±0.5 4.0±0.4 d7 Comparative Example 1 5.4±0.5 4.4±0.4 d7 Example 1 Group 4.5±0.5 3.6±0.4

[0106] The results showed that the Example 1 group exhibited the lowest pathogen load at both time points.

[0107] This application introduces Aspergillus niger fermentation technology to biotransform traditional Chinese medicine compound, and systematically evaluates its prevention and control effects on three typical bacterial diseases—pullorum disease, colibacillosis, and fowl cholera—at two key production stages: chicks and growers. The results show that, regardless of the chick or grower stage, Example 1 exhibits stable and consistent advantages in multiple key endpoints, including reducing morbidity and mortality, shortening disease duration, improving growth performance, and reducing bacterial load. Its overall effect is superior to Comparative Example 1, and it is superior to or not inferior to florfenicol in most indicators.

[0108] From the perspective of disease type, the three bacterial diseases selected in this application differ to some extent in their pathogenesis and clinical manifestations: pullorum disease and colibacillosis are mainly characterized by intestinal infection and systemic dissemination, while fowl cholera is more inclined towards acute septicemic infection. However, the results of this application show that Example 1 can significantly reduce bacterial load and improve clinical outcomes in all three different types of bacterial disease models, suggesting that the preparation is not only effective against a single pathogen or a specific type of infection, but also has a certain broad-spectrum prevention and control potential; this is of great significance for the complex situation of "multiple pathogens mixed or rotating epidemics" in actual production.

[0109] From the perspective of age stages, this application conducted a systematic evaluation on both chicks and pullets, two key production stages. The results showed that Example 1 consistently demonstrated a stable preventative effect in both chicks, whose immune systems are not yet fully mature, and pullets, which experience rapid growth and a heavy metabolic burden. Furthermore, it significantly reduced bacterial load and improved production performance at both stages. This indicates that the application of this formulation is not significantly limited by the physiological stage of the chicken, demonstrating good age applicability and potential for widespread application.

[0110] It is noteworthy that this application not only evaluated the prevention and control efficacy from the perspectives of clinical symptoms and production performance, but also systematically introduced bacterial load as a pathogen evaluation indicator. The results showed that in all combinations of "disease and age," Example 1 significantly reduced the pathogen load in feces and target organs at different time points after challenge, and in most cases, its reduction was superior to Comparative Example 1, and equal to or better than florfenicol. This result indicates that the formulation can not only alleviate clinical symptoms, but more importantly, it can inhibit bacterial colonization and systemic dissemination at the pathogen level, thereby providing a basis for reducing the risk of recurrence and improving the health status of the population.

[0111] Compared with Comparative Example 1, Example 1 showed more stable and significant prevention and control effects in all models, suggesting that the Aspergillus niger fermentation process plays a key role in improving the overall efficacy of the preparation. Although this application does not provide an in-depth analysis of the specific material basis, given that Aspergillus niger is known to produce a variety of hydrolytic enzymes and promote the release of active ingredients in traditional Chinese medicine, it is reasonable to speculate that the fermentation process may improve the bioavailability of active ingredients in traditional Chinese medicine, improve the palatability of the preparation, or promote the optimization of the intestinal environment through multiple pathways. Furthermore, the specific traditional Chinese medicine compound formulation in this application synergistically enhances the overall prevention and control effect.

[0112] A comparison with florfenicol reveals that florfenicol has a relatively clear effect on controlling acute symptoms and reducing mortality in the early stages after challenge, but its administration time is usually short and it is mainly positioned as a "therapeutic drug." In contrast, Example 1, used continuously as a feed additive, not only showed comparable or better clinical outcomes than florfenicol in multiple models, but also demonstrated a more stable overall advantage in reducing bacterial load and improving growth performance. This characteristic of "continuous regulation + stable inhibition" makes it more suitable for long-term prevention and control and antibiotic reduction and replacement in large-scale farming.

[0113] In summary, this application systematically demonstrates the stability and broad-spectrum advantages of Example 1 in the prevention and control of bacterial diseases in chickens from three dimensions: multiple diseases, multiple ages, and multiple levels of indicators. Compared with the traditional Comparative Example 1 and conventional antibiotics, this formulation has better comprehensive application potential under actual production conditions, and is especially suitable for long-term addition to reduce the risk of disease outbreaks in flocks and improve overall production performance.

[0114] The embodiments presented herein are merely selected implementations based on combinations of all possible embodiments. The appended claims should not be limited to the embodiments described herein. Some numerical ranges used in the claims include sub-ranges within them, and variations within these ranges should also be covered by the appended claims.

Claims

1. A fermented traditional Chinese medicine compound, characterized in that, It is obtained by fermentation of traditional Chinese medicine compound with Aspergillus niger. The traditional Chinese medicine compound comprises the following components by weight: 70-90 parts Coptis chinensis, 90-110 parts Scutellaria baicalensis, 110-130 parts Lonicera japonica, 90-110 parts Forsythia suspensa, 110-130 parts Pulsatilla chinensis, 90-110 parts Andrographis paniculata, 110-130 parts Taraxacum mongolicum, 70-90 parts Atractylodes lancea, 90-110 parts Atractylodes macrocephala, 90-110 parts Poria cocos, and 30-50 parts Glycyrrhiza uralensis.

2. A method for preparing a fermented traditional Chinese medicine compound as described in claim 1, characterized in that, Includes the following steps: Step 1: Prepare the traditional Chinese medicine compound into a matrix; Step 2: Inoculate the substrate with Aspergillus niger for fermentation to obtain the fermented traditional Chinese medicine compound.

3. The method for preparing fermented traditional Chinese medicine compound according to claim 2, characterized in that, The specific operation of step 1 is as follows: Coptis chinensis, Scutellaria baicalensis, Lonicera japonica, Forsythia suspensa, Pulsatilla chinensis, Andrographis paniculata, Taraxacum mongolicum, Atractylodes lancea, Atractylodes macrocephala, Poria cocos and Glycyrrhiza uralensis are pulverized into powder, and then mixed with 8 times the weight of the powder with water to prepare a moist matrix. The moist matrix is ​​then sterilized under high pressure at 121°C for 25 minutes and cooled to 32°C to obtain the matrix.

4. The method for preparing fermented traditional Chinese medicine compound according to claim 2, characterized in that, The specific operation of step 2 is as follows: Inoculate the substrate with spores at a concentration of 1×10⁻⁶ at a wet weight ratio of 1% (v / w). 7 CFU / ml is a suspension of Aspergillus niger spores; then solid-state fermentation is carried out at 30-33℃ for 72 hours to obtain fermented traditional Chinese medicine compound.

5. The method for preparing fermented traditional Chinese medicine compound according to claim 4, characterized in that, The specific process of solid-state fermentation is as follows: the relative humidity of the environment is maintained at 60-70%, and the initial pH is controlled at 5.0-6.0; the material is turned over once every 24 hours during fermentation.

6. The method for preparing fermented traditional Chinese medicine compound according to claim 4, characterized in that, It also includes step 3: drying the fermented Chinese herbal medicine compound at 60°C until the moisture content is less than 10%, then pulverizing and passing it through a 40-mesh sieve.

7. Application of drugs for the prevention and treatment of bacterial diseases in poultry prepared using the fermented traditional Chinese medicine compound as described in claim 1.

8. Application of feed additives for the prevention and treatment of bacterial diseases in poultry prepared using the fermented traditional Chinese medicine compound as described in claim 1.