2 kinds of anti-viral compound traditional Chinese medicines based on ma-xing-shi- gan decoction for chicken infectious bronchitis and preparation method and application thereof

Abstract

The application discloses two kinds of anti-virus compound traditional Chinese medicines for chicken infectious bronchitis based on the increase and decrease of Maxingshigan decoction, and a preparation method and application thereof, and relates to the technical field of veterinary compound traditional Chinese medicines. The two kinds of anti-virus compound traditional Chinese medicines are prepared into 1 g / ml oral liquid according to the weight ratio of 9 parts of liquorice, 9 parts of bitter almond, 9 parts of platycodon grandiflorum, 9 parts of ephedra, 14 parts of fritillaria; 9 parts of platycodon grandiflorum, 7 parts of stemona, 9 parts of liquorice, 7 parts of Imonium, 9 parts of ephedra and 9 parts of bitter almond, and are given to chicken for free drinking water at 3.6 ml / L for 7 days in succession. The application is simple to prepare, can realize mass production, can effectively treat chicken infectious bronchitis virus, greatly reduces the infection and disease course of chicken infectious bronchitis virus disease of chicken, reduces the mortality of sick chickens, reduces drug residues, improves the production performance of chicken, and creates higher economic benefits.

Description

Technical Field

[0001] This invention relates to the field of compound traditional Chinese medicine for veterinary use, and particularly to two compound traditional Chinese medicines for the prevention and treatment of infectious bronchitis in chickens, their preparation methods, and applications. Background Technology

[0002] Infectious bronchitis (IB) is an acute, highly contagious disease of chickens caused by the infectious bronchitis virus (IBV), a virus belonging to the genus *Gammacoronavirus* of the family Coronaviridae. IBV is a single-stranded, positive-sense RNA virus containing four structural proteins: spike (S) protein, membrane (M) protein, nucleocapsid (N) protein, and small membrane (E) protein. IBV infection in chickens is primarily characterized by pathological changes in the respiratory tract, kidneys, and reproductive system. Affected chickens exhibit tracheal rales, coughing, sneezing, and decreased egg production and quality, causing significant economic damage to the poultry industry. Currently, IBV is mainly prevented using attenuated or inactivated vaccines. However, due to the multiple IBV serotypes and the lack of effective cross-protection between different serotypes, new variant strains are constantly emerging, posing a significant challenge to the diagnosis and control of this disease.

[0003] In the context of antibiotic-free farming and the lack of effective antiviral Western medicines in animal husbandry, traditional Chinese medicine (TCM) has become the main antiviral drug for livestock. TCM herbs inhibit the reproduction of pathogenic microorganisms by interfering with their normal metabolism, suppressing their nucleic acid and protein synthesis, or enhancing the animal's immune system. This directly or indirectly inhibits or kills pathogenic microorganisms, demonstrating good preventative and therapeutic effects against viral diseases in livestock and poultry. Furthermore, TCM herbs are characterized by low residue, low toxicity, high cure rates, and low likelihood of inducing drug resistance, thus meeting the requirements of green farming.

[0004] According to traditional Chinese medicine theory, IBV in chickens is caused by pathogenic factors invading the lungs, leading to obstruction of qi and blood circulation, resulting in impaired lung function. Therefore, the root cause of IBV lies in the lungs. Traditional Chinese veterinary medicine treatment for IBV should follow the two principles of "expelling pathogens" and "strengthening the body's resistance," primarily using expectorant and antiasthmatic herbs to eliminate pathogenic factors and enhance the body's antiviral capabilities, thereby improving immune function. In recent years, the use of traditional Chinese medicine, its active ingredients, and compound formulas to prevent and treat IBV has achieved some success. However, most of these methods involve simple screening of known antiviral drugs and comparative experiments to identify relatively advantageous herbs. Such experimental processes and methods cannot scientifically and effectively determine truly effective drugs against IBV. Therefore, a more precise method of summarizing drug patterns and logically deducing suitable compound formulas is needed. Summary of the Invention

[0005] This invention aims to provide two compound traditional Chinese medicine formulas based on modified Ma Xing Shi Gan Tang that can prevent and treat infectious bronchitis (IBV) in chickens, alleviating the predicament of the complete ban on antiviral chemical drugs in livestock and poultry farming, and has good economic and social benefits.

[0006] Another objective of this invention is to provide a method for preparing the above-mentioned compound traditional Chinese medicine, wherein the preparation is a compound water extract. By selecting appropriate medicinal materials and their proportions and formulating the preparation method, the method is green and safe, simple to operate, and has significant therapeutic effects.

[0007] Another object of the present invention is to provide the clinical application of the above-mentioned compound traditional Chinese medicine in the safe and effective treatment and prevention of IBV.

[0008] The weight units mentioned in this invention can be μg, mg, g, kg, or other weight units known in the pharmaceutical field.

[0009] The infectious bronchitis virus described in this invention is a respiratory infectious bronchitis virus, and the strain is M41. This virus infection is a single infection.

[0010] The chicken described in this invention is the Guangxi Tu Sanma Chicken.

[0011] To achieve the above objectives, the present invention adopts the following technical solution:

[0012] Based on preliminary exploration and analysis of traditional Chinese medicine theory, and referring to the ratio of licorice and bitter apricot kernel in the Ma Xing Shi Gan Tang formula of the Veterinary Pharmacopoeia, the optimal ratio was calculated based on the formula calculation platform of traditional Chinese medicine inheritance. The optimal ratio of the compound Chinese medicine is prepared from the following Chinese medicinal materials in the following weight parts: licorice 9 parts, bitter apricot kernel 9 parts, platycodon 9 parts, ephedra 9 parts, fritillaria 14 parts; platycodon 9 parts, stemona 7 parts, licorice 9 parts, cynanchum 7 parts, ephedra 9 parts, bitter apricot kernel 9 parts.

[0013] The preparation method of the compound traditional Chinese medicine is as follows:

[0014] Take Chinese medicinal herbs, chop them separately using an automatic slicer, mix them according to the formula ratio, soak them in cold water 2-5 cm above the herbs for 20-30 minutes, bring to a boil over high heat, then simmer over low heat for 20-30 minutes. After decocting, pour out the liquid, add water to the dregs and decoct twice more using the same method, but for shorter times. After mixing the liquids three times, centrifuge at 3000 rpm for 15 minutes to remove the herb residue and collect the supernatant. Concentrate the supernatant using a rotary evaporator to a quantitative ratio of 1g / ml to prepare the compound Chinese medicine water extract.

[0015] Furthermore, the prepared compound herbal extract is made into an oral liquid and administered by adding it to drinking water.

[0016] Furthermore, the dosage of the medication in the drinking water is 2.4-3.6 ml / L, and the chickens have free access to the water.

[0017] Furthermore, the compound traditional Chinese medicine is used to treat respiratory diseases in chickens caused by infectious bronchitis virus.

[0018] This invention is based on traditional Chinese veterinary medicine theory, the characteristics of Chinese herbal medicines, and the pathogenesis of viral diseases in livestock and poultry. It follows the strict formulation of traditional Chinese medicine according to the principles of monarch, minister, assistant, and guide. The combination of these herbs has the effects of resolving phlegm, relieving cough and asthma, clearing heat and tonifying deficiency, and relieving exterior syndromes and regulating qi. Each herb has a clear division of labor and performs its own function. This invention is a compound Chinese medicine for resisting IBV and its preparation method, which has been screened one by one in clinical trials.

[0019] Regarding the clinical application of the aforementioned compound traditional Chinese medicine, this invention uses Guangxi Ma chicken (Tu San) as the experimental subject. By constructing a respiratory disease model in chicks caused by IBVM41 strain, the effects of the drug of this invention on the respiratory disease model in chicks caused by IBVM41 strain were studied from the perspectives of chick clinical condition, blood routine test, and viral load test of various organs, thus confirming the effectiveness of the drug of this invention. Attached Figure Description

[0020] Figure 1 These are the high-performance liquid chromatograms of the two compound preparations obtained in this invention and Ma Xing Shi Gan Tang;

[0021] Figure 2 This is a clinical evaluation of the two compound preparations obtained in this invention after in vivo treatment of infectious bronchitis in chickens, and a graph showing the average daily weight gain of the chickens.

[0022] Figure 3 This is a comparative graph of immune organ indices in chicks after in vivo treatment with two compound preparations obtained in this invention for infectious bronchitis in chickens.

[0023] Figure 4 This is a graph showing the viral load detection after in vivo treatment of infectious bronchitis in chickens with the two compound preparations obtained in this invention.

[0024] Figure 2 (A) Clinical evaluation; (B) Average daily weight gain;

[0025] Figure 3 (A) Thymus; (B) Spleen; (C) Bursa of Fabricius;

[0026] Figure 4 In the Chinese dictionary: (A) nasal swab; (B) trachea; (C) lung; (D) kidney. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0028] Example 1: Generation of Traditional Chinese Medicine Compound Formulas

[0029] This embodiment constructs a dataset targeting the symptoms of infectious bronchitis in chickens and uses it to generate two modified Ma Xing Shi Gan Tang formulas that are theoretically effective against infectious bronchitis virus in chickens.

[0030] Using data from 34,324 traditional Chinese medicine (TCM) formulas in the Chinese Formula Database, 7,505 TCM compound preparations and 33,837 traditional formulas in the cross-reference database of traditional Chinese medicine formulas, and 4,207 TCM compound preparations in the TCM Inheritance Computing Platform, and searching the Chinese Veterinary Pharmacopoeia for keywords such as "cough," "lung," "toxin," and "phlegm," a total of 1,121 accurately recorded TCM formulas were retrieved from the Chinese Formula Database, 853 TCM compound preparations and 1,102 traditional formulas from the cross-reference database of traditional Chinese medicine formulas, 464 compound preparations from the 4,207 TCM compound preparations dataset of the TCM Inheritance Computing Platform, 11 compound preparations from the Chinese Veterinary Pharmacopoeia, and 11 new veterinary drug formulas were retrieved. Currently, among the existing compound preparations, those similar to those related to the occurrence of infectious bronchitis virus in chickens can be adopted. The above compound preparation data were analyzed using high-frequency combinations of formulas and k-means clustering analysis using the "TCM Inheritance Computing Platform V3.0" software. Strictly following the above criteria, a total of 353 prescriptions for treating cough were selected, involving 361 kinds of drugs, and a database for treating infectious bronchitis in chickens was constructed based on these prescriptions. Cluster analysis was performed on the 353 prescriptions and 361 Chinese medicinal herbs included, and the modified Ma Xing Shi Gan Tang prescription was automatically generated as shown in Table 1 below.

[0031] Table 1 Analysis of Core Drug Combinations

[0032]

[0033] Example 2: Preparation of Traditional Chinese Medicine Compound Oral Liquid

[0034] This study obtained an effective compound antiviral oral liquid through water extraction, and the preparation method is as follows:

[0035] Weigh out the following Chinese medicinal herbs: 90g of licorice root, 90g of bitter almond, 90g of platycodon root, 90g of ephedra, and 140g of fritillaria bulb; and 90g each of platycodon root, stemona root, licorice root, cynanchum root, ephedra, and bitter almond. Chop each herb separately using an automatic slicer and then mix them to obtain two 500g compound herbal formulas. Place the above 500g herbs in a clay pot, add cold water to cover the herbs by 2-5 cm, soak for 20-30 minutes, then bring to a boil over high heat, and simmer over low heat for 20-30 minutes. Pour out the decoction and set aside. Add water to the dregs and decoct twice more using the same method, but for shorter times. Combine the three decoctions. Divide the decoction into 100ml centrifuge tubes and centrifuge at 3000 rpm for 15 minutes to remove the herbal residue and collect the supernatant. The supernatant was concentrated to a drug concentration of 1 g / ml using a rotary evaporator. After being dispensed and placed in 65°C warm water for 30 minutes to achieve pasteurization, it was then stored in a 4°C refrigerator for later use.

[0036] Based on the two compound formulas obtained in this invention and the use of Ma Xing Shi Gan Tang, high-performance liquid chromatography (HPLC) was performed using an Aglent 1290 ultra-high performance liquid chromatography tandem with Thermo Fisher Scientific Q-Exactive Orbitrap Plus high-resolution mass spectrometry. Figure 1 The testing process is as follows:

[0037] 1. Chromatographic conditions: Wates T3 (21mm×50mm, 18μm), column temperature 50℃; A is 0.1% formic acid solution; B is acetonitrile; flow rate 0.3mL / min; 0-1min 2%; 1-18min 100%; 18-22min 100%; 22-22.1min 2%; 22.1-25min 2%; injection volume 5μL.

[0038] 2. Set the mass spectrometry conditions: DDA is 100-1000 mz; ESI source conditions are set as follows: auxiliary gas flow rate is 16 Arb, Full ms resolution is 70000, collision energy in NCE model is 25 eV, MS / MS resolution is 17500, and spray voltage is -3.0 kV (negative) or 3.0 kV (positive).

[0039] 3. Detection process: Dynamic exclusion for 4 seconds, energy step NCE, 10, 30, 50, data acquisition range m / z 100-1000, full scan mode, data processing using Xcabur 4.3 and MS-DIAL 5.0.3 workstations, compound identification by comparison with reference standards, retrieval from a local self-built database and diagnostic ion confirmation, combined with the fragmentation patterns of primary quasi-separators and secondary mass spectrometry.

[0040] Example 3: Treatment Experiment for Artificial IBV Infection

[0041] This study investigates the therapeutic effect of the present invention on IBV through experiments. The verification process is as follows:

[0042] 1. Experimental Design

[0043] 14-day-old Guangxi Ma chickens (local breed 3); 0.2 ml of highly virulent strain M41 of infectious bronchitis virus (IBV) containing 10... 5 EID 50 Except for the blank control group, the other groups were divided into 10 groups. 5 EID 50 0.2 ml of the herbal compound oral liquid was administered via eye drops and nasal drops for viral attack. The traditional Chinese medicine compound oral liquid prepared in Example 2 was purchased from Luoyang Huizhong Veterinary Medicine Co., Ltd. (Production batch number: 20220602; Approval number: Veterinary License No. 160036112; Specification: 2.4 g / ml), and ribavirin was purchased from Shanxi Taiyuan Co., Ltd. (Approval number: National Medicine Approval No. H14020252; Specification: 0.1 g / ml). The experiment was divided into 8 groups: virus infection group, blank control group, Huizhong Gankang control group (high-dose and medium-dose, i.e., the positive control group of the traditional Chinese medicine Ma Xing Shi Gan Tang), and the two traditional Chinese medicine compound groups prepared in Example 2 (high-dose and medium-dose), with 30 Guangxi Ma chickens in each group. The control group received Huizhong Gankang oral solution (2.4g / ml), at medium doses of 1ml / L and high doses of 1.5ml / L (according to the instructions). The traditional Chinese medicine compound group received traditional Chinese medicine compound oral solution (1g / ml), at medium doses of 2.4ml / L and high doses of 3.6ml / L (referring to the dosage of Huizhong Gankang). All chickens were given the medication via free access to water during the study. The medication was administered simultaneously with the viral load, diluted in 1L of water each time. To prevent water contamination from chicken feces, 500mL was added twice daily, morning and evening, for 7 consecutive days.

[0044] 2. Clinical symptoms and daily weight gain

[0045] Criteria for judging the efficacy of compound traditional Chinese medicine in prevention and treatment (Table 2):

[0046] Table 2. IBV Experimental Clinical Evaluation Scores

[0047]

[0048]

[0049] Clinical symptoms and daily weight gain results indicated that symptom scores significantly decreased in all TCM treatment groups after IBV infection, while symptom scores remained persistently high in the virus-treated groups. A significant decrease in symptom scores was observed in all treatment groups at the 3-day post-infection (dpi) marker, indicating a critical inflection point. Among these groups, the MXSG-mix2-H group had the lowest symptom score at 5 dpi. By the 14-day post-infection (dpi) marker, symptom scores in all TCM treatment groups, except for the MXSG-M group, had decreased to zero. Figure 2 A). In terms of growth performance, both dose levels in the MXSG-mix2 group were superior to other treatment groups. The growth curves of this group were very similar to those of the control group. Analysis of the trend of the curve changes shows that the MXSG-mix2-H group demonstrated the most effective clinical performance in combating the effects of IBV infection. Figure 2 B).

[0050] 3. Tracheal score

[0051] One day after drug withdrawal (7 dpi), five chickens from each group were euthanized. Under aseptic conditions, the trachea was extracted and divided into three loops in the upper section, four loops in the middle section, and three loops in the lower section for evaluating ciliary activity. The remaining middle segment of the trachea was fixed in 10% formalin, embedded in paraffin at 60°C, and then sectioned into 5-μm thick sections. The sections were dewaxed, dehydrated using a gradient of alcohols, and then stained with H&E. The treated sections were dehydrated using a gradient of alcohols, dewaxed, sealed with coverslips, and observed under a microscope. The ciliary movement scores of the five chickens were averaged, and a protection score was calculated (protection score = [1 - average score of the treatment group / average score of the infection group] × 100%).

[0052] Tracheal scoring results showed that at 7 dpi, 5 chickens in each group were euthanized for further examination. Their tracheae were extracted to form tracheal rings, and the health of the cilia was assessed under an optical microscope. The results, as shown in Table 3, indicated that the IBV group, in severe cases, exhibited excessive tracheal mucus, impaired ciliary movement, and ciliary loss. The average tracheal ciliary survival rate score in this group was 38.5 points, indicating no protective effect. In contrast, the tracheal protection rate in the MXSG-mix group exceeded 60%, with the highest protection rate (70.62%) observed in the MXSG-mix2-H group. Conversely, the protection rate in the MXSG group remained below 60%.

[0053] Table 3. Ciliary body scoring results

[0054]

[0055]

[0056] 4. Blood routine test results

[0057] Simultaneously, 1 mL of blood was drawn from the arm vein. An anticoagulant was added, and routine blood parameters were measured using a fully automated blood analyzer (TEK-VET3, TECOM, Jiangxi, China). These parameters included total white blood cell count (WBC), total red blood cell count (RBC), hemoglobin content (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW), platelet count (PCT), mid-intensity index (MID%), lymphocyte percentage (LYM%), and granulocyte percentage (GRAN%).

[0058] The complete blood count (CBC) data are shown in Table 4. The results indicate that at 7 dpi, venous blood samples were collected from 5 chickens in each treatment group for complete blood count (CBC) testing. The results showed that the effects of different traditional Chinese medicine compound formulas on chicken blood parameters varied depending on the collection time. When comparing each treatment group with the challenge group, MXSG-mix1 treatment significantly affected certain blood parameters, including white blood cell count (WBC) at 3 dpi, hematocrit (HCT) at 14 dpi, mean erythrocyte volume (MCV) at 3 dpi, and platelet distribution width (PDW) at 3 dpi. In group H, platelet count (PCT) at 3 dpi, as well as lymphocyte percentage (LYM%) and granulocyte percentage (GRAN%) at 3 and 7 dpi, were also affected. MXSG-mix2 treatment significantly affected HCT at 14 dpi in group M, MCV and PDW at 3 dpi in group M, and the percentage of intermediate-dip cells (MID%) at 3 dpi in group M and 7 dpi in group H. At 7 days post-contraction (dpi), the effect on GRAN% in group H was also significant. In group MXSG-H, at 14 days post-contraction (dpi), significant effects were observed on HCT, MCV, mean corpuscular hemoglobin (MCH), platelet count (PLT), mean platelet volume (MPV), MID% at 7 days post-contraction (dpi), and LYM at 7 days post-contraction (dpi). In summary, most blood parameters began to show the effects of traditional Chinese medicine treatment at 3 days post-contraction (dpi), and some effects persisted for one week after discontinuation of treatment. Overall improvement in health status was comparable to the control group.

[0059] Table 4 Blood Routine Examination

[0060]

[0061]

[0062] 5. Immune organ index status

[0063] Five chickens were treated on day 3 (3 dpi), day 1 (7 dpi) after drug withdrawal, and day 7 (14 dpi) after drug withdrawal. Spleen, thymus, and bursa of Fabricius were collected from each group to calculate the immune organ index.

[0064] A comparison chart of immune organ indices is shown below. Figure 3 As shown, the results indicate that comprehensive analysis at specific time intervals (3 dpi, 7 dpi, and 14 dpi) after infection is effective in assessing immune organ indices, with a focus on key organs such as the spleen, thymus, and alveolar sacs. Five chickens were selected from each group, and the weight of each organ was measured to calculate the relative organ weight. Figure 2 At 3 dpi, the thymus index in the MXSG-mix2-M group was significantly elevated, indicating a significant difference from the challenge group. At 7 dpi, except for the MXSG-M group, most TCM treatment groups showed significant differences from the challenge group. The difference in the MXSG-H group was the most significant, remaining consistent until 14 dpi. Throughout the experiment, the spleen index in the treatment groups consistently exceeded that in the challenge group. However, it is noteworthy that no significant difference was observed between the MXSG-mix2-M group and the control group during this period. Regarding the bursa of Fabricius index, the IBV group showed a significant decrease at 3 dpi. However, compared to the challenge control group, each drug treatment group showed a significant increase in the bursa of Fabricius index. By 7 dpi, both the MXSG-mix1-H and MXSG-mix2 groups maintained high vesicular sac indices. By 14 dpi, the difference between the two groups became insignificant.

[0065] 6. Viral load measurement results

[0066] On days 3, 8, and 15 of the experiment, five chickens from each group were randomly selected for dissection. Trachea were collected and placed in 1.5 EP tubes containing 1 ml of RNA stabilizing solution (Genstar, catalog number P301-10). Nucleic acid was extracted using a column-based viral DNA / RNA extraction kit (Genstar, catalog number P142-01). Viral RNA was reverse transcribed into cDNA using a reverse transcription kit (Genstar, catalog number A232-02). The viral load in the tracheal tissue was determined by fluorescent PCR using primers and probes from a commercial IBV virus detection kit (NEQ13700-T, Harbin Guosheng Biotechnology Co., Ltd., Harbin, China). The premixed enzyme used for the fluorescent PCR was provided by Takara Bio Inc. (catalog number: A039). The reverse transcription procedure was strictly performed according to the manufacturer's instructions. The total volume for the quantitative real-time PCR (probe method) was 20 μL, and the reaction program was as follows: 10 μL Premix Ex Taq™ (Perfect Real Time), 0.4 μL each of IBV-F (20 pmol / μL) and IBV-R (20 pmol / μL), 0.3 μL of IBV-P (20 pmol / μL), 2 μL of template, and 6.9 μL of RNase-free water. The amplification program was: pre-denaturation at 95℃ for 30 s; denaturation at 95℃ for 5 s; annealing at 62℃ for 34 s, for 40 cycles, during which fluorescence signals were collected simultaneously.

[0067] The viral load measurement graph is shown below. Figure 4 As shown, the results indicated that no virus was detected in the trachea and kidneys of chickens in the NC group. However, the expression level of IBV mRNA in the trachea and kidneys of the IBV group was significantly higher than that of the treatment group. The study found that the viral load in nasal swabs and trachea was higher than that in the lungs and kidneys. The Traditional Chinese Medicine (TCM) group significantly reduced viral invasion of the lungs at 7 days post-contraction (dpi). At 3 dpi, the MXSG-mix1-H group showed stronger antiviral activity in the respiratory tract. On the other hand, at 7 dpi, the MXSG-mix2-H group showed enhanced antiviral activity in the respiratory tract. Throughout the experiment, except for lung indicators at 3 dpi, the MXSG-mix-H group showed stronger antiviral effects than the MXSG-H group. By 14 dpi, the viral load in all groups began to decrease, but the decrease was more pronounced in the samples treated with TCM. The MXSG-mix2-H group recorded the lowest viral load, almost equivalent to the blank value for each organ.

[0068] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. 2 kinds of antiviral compound traditional Chinese medicine based on the modified Ma Xing Shi Gan Tang formula for infectious bronchitis in chickens, characterized in that, The compound composition obtained by optimizing the formula of Ma Xing Shi Gan Tang consists of licorice, bitter apricot kernel, platycodon, ephedra, and fritillaria; platycodon, stemona, licorice, cynanchum, ephedra, and bitter apricot kernel.

2. The two antiviral compound traditional Chinese medicine formulas for infectious bronchitis in chickens based on the modified Ma Xing Shi Gan Tang formula according to claim 1, characterized in that, The effective ingredients are prepared from the following Chinese medicinal materials in parts by weight: 9 parts licorice, 9 parts bitter almond, 9 parts platycodon, 9 parts ephedra, 14 parts fritillaria; 9 parts platycodon, 7 parts stemona, 9 parts licorice, 7 parts cynanchum, 9 parts ephedra, and 9 parts bitter almond.

3. A method for preparing two antiviral compound traditional Chinese medicine formulas for infectious bronchitis in chickens based on modified Ma Xing Shi Gan Tang according to claim 1 or 2, characterized in that, Includes the following steps: (1) Weigh out the Chinese medicinal materials according to the weight proportions and then chop them separately using an automatic slicer. Mix the various Chinese medicinal materials according to the formula ratio. (2) Soak the herbs in cold water for 2-5 cm above the herbs for 20-30 minutes. Then bring to a boil over high heat and simmer over low heat for 20-30 minutes. After decocting, pour out the liquid. Add water to the dregs and decoct again using the same method as the first time, but for a shorter time. Then mix the decocted liquid with the herbs. (3) Centrifuge to remove the residue of Chinese medicine and take the supernatant; (4) The extract is concentrated to a certain ratio of drug solution by rotary evaporator to prepare compound Chinese medicine water extract.

4. The preparation method of the two antiviral compound traditional Chinese medicines for infectious bronchitis in chickens based on the modified Ma Xing Shi Gan Tang formula according to claim 3, characterized in that, In step (2), the dregs of the Chinese medicine are decocted twice with water in the same way as the first decoction.

5. The preparation method of the two antiviral compound traditional Chinese medicines for infectious bronchitis in chickens based on the modified Ma Xing Shi Gan Tang formula according to claim 3, characterized in that, In step (2), the decoctions from the three decoctions are mixed.

6. The preparation method of the two antiviral compound traditional Chinese medicines for infectious bronchitis in chickens based on the modified Ma Xing Shi Gan Tang formula according to claim 3, characterized in that, In step (3), the herbal residue is removed by centrifugation at 3000 rpm for 15 minutes to obtain the supernatant.

7. The preparation method of the two antiviral compound traditional Chinese medicines for infectious bronchitis in chickens based on the modified Ma Xing Shi Gan Tang formula according to claim 3, characterized in that, In step (4), the ratio of the medicinal liquid is 1g / ml.

8. The application of an antiviral compound traditional Chinese medicine prepared according to any one of claims 3-7, characterized in that, The prepared compound herbal extract is an oral liquid that is added to drinking water for administration.

9. The application of the antiviral compound traditional Chinese medicine according to claim 8, characterized in that, The concentration of the compound herbal extract is 1 g / ml, and the dosage in drinking water is 2.4-3.6 ml / L, with free access to drinking water for chickens.

10. The application of the antiviral compound traditional Chinese medicine according to claim 8, characterized in that, The aforementioned antiviral compound traditional Chinese medicine is used to treat respiratory diseases in chickens caused by infectious bronchitis virus.

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