A traditional Chinese medicine composition for preventing and treating pig bacterial pneumonia, a preparation method and application thereof

Abstract

The application discloses a traditional Chinese medicine composition for preventing and treating pig bacterial pneumonia and a preparation method and application thereof. The traditional Chinese medicine composition comprises the following components: Ephedra, apricot kernel, gypsum, Astragalus root, Anemarrhena asphodeloides, Cape jasmine fruit, talc, licorice, honeysuckle, forsythia, rhubarb, mirabilite, peony root and kudzurpini radix. The application studies and clinically applies traditional Chinese medicine by combining traditional preparation technology with modern preparation technology, and adopts the traditional Chinese medicine compound prepared through fermentation to prevent and treat pig bacterial pneumonia, so as to solve the above problems. Through clinical application, the traditional Chinese medicine compound has a treatment effect on pig bacterial pneumonia, and the cure rate can reach 91.83%, and the cure rate can reach 95.97% after fermentation, the drug efficacy is improved, the recurrence is difficult, and there is no drug residue. In addition, the components are common traditional Chinese medicines, the cost is low, the safety is high, and there is no side effect. The application not only reduces the economic loss of breeders, but also meets the demand of people for green food, and conforms to the development trend.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, and in particular relates to a traditional Chinese medicine composition for the prevention and treatment of bacterial pneumonia in pigs, its preparation method and application. Background Technology

[0002] Respiratory diseases are a major concern in intensive pig farming. Porcine respiratory syndrome (PRS) is a highly fatal disease resulting from mixed infections of multiple bacteria. Stressful factors such as high temperatures, humidity, and heat promote its occurrence and spread, causing significant economic losses to pig farms.

[0003] Common respiratory infections caused by bacteria include Haemophilus parasuis infection, swine pneumonia, and swine infectious pleuropneumonia. Airborne transmission is the primary route of infection, with airborne aerosol transmission being the main mode of transmission. Pigs of all ages are susceptible, especially weaned piglets and nursery piglets. After infection, bacterial toxins produce pyrogens, stimulating the thermoregulatory center and causing an increase in body temperature. Simultaneously, bacterial toxins can cause bronchitis and pneumonia, manifesting as difficulty breathing and coughing. Pneumonia can cause tissue hypoxia, peripheral circulatory failure, purplish or blackened discoloration of the ear tips and subcutaneous tissue of the abdomen, and even necrosis. Endotoxic shock can exacerbate these symptoms, and circulatory disturbances can lead to edema. Therefore, during the infection period, treatment with traditional Chinese medicine, which has fewer side effects, is more effective than the extensive use of Western medicine.

[0004] Currently, the prevention and treatment of bacterial pneumonia primarily rely on vaccination and broad-spectrum antibiotics such as tilmicosin, norfloxacin, kanamycin, and ceftriaxone. However, due to the large number of bacterial serotypes, it is impossible to select a vaccine that corresponds to the locally prevalent strain. Large doses and prolonged use of broad-spectrum antibiotics can increase the burden on the liver and kidneys, severely affecting efficacy, and they also commonly exhibit drug resistance, are prone to relapse, and easily leave residues in the body. Furthermore, group treatment often employs mixed feeding methods. Pigs have a sensitive sense of smell and taste; adding bitter or astringent drugs will reduce feed intake, affecting drug efficacy.

[0005] As people's demand for green livestock products increases, traditional Chinese medicine, with its advantages of wide availability, significant efficacy, no residue, and few toxic side effects, is gaining more and more attention as an alternative to chemical drugs in the prevention and treatment of livestock and poultry diseases, and has broad market prospects. Summary of the Invention

[0006] The purpose of this invention is to provide a traditional Chinese medicine composition for the prevention and treatment of bacterial pneumonia in pigs, its preparation method, and its application.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] In the first aspect, the present invention provides a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components by weight: 40-60 parts of ephedra, 10-20 parts of apricot kernel, 80-120 parts of gypsum, 40-60 parts of astragalus, 40-60 parts of anemarrhena, 40-60 parts of gardenia, 30-50 parts of talc, 10-20 parts of licorice, 30-50 parts of honeysuckle, 10-20 parts of forsythia, 10-15 parts of rhubarb, 10-15 parts of mirabilite, 30-50 parts of angelica dahurica, and 40-60 parts of kudzu root.

[0009] Preferably, the ingredients include the following components by weight: 60 parts ephedra, 20 parts apricot kernel, 120 parts gypsum, 60 parts astragalus, 60 parts anemarrhena, 60 parts gardenia, 50 parts talc, 20 parts licorice, 50 parts honeysuckle, 20 parts forsythia, 15 parts rhubarb, 15 parts mirabilite, 50 parts angelica, and 60 parts kudzu root.

[0010] Secondly, this invention provides another traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components by weight: 40-60 parts ephedra, 10-20 parts apricot kernel, 80-120 parts gypsum, 40-60 parts astragalus, 40-60 parts anemarrhena, 40-60 parts gardenia, 30-50 parts talc, 10-20 parts licorice, 30-50 parts honeysuckle, 10-20 parts forsythia, 10-15 parts rhubarb, 10-15 parts mirabilite, 30-50 parts angelica, and 40-60 parts kudzu root; and is prepared by fermentation.

[0011] Thirdly, the present invention provides a method for preparing a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs as described in the second aspect above, comprising the following steps:

[0012] Ephedra, apricot kernel, gypsum, astragalus, anemarrhena, gardenia, talc, licorice, honeysuckle, forsythia, rhubarb, mirabilite, angelica, and kudzu root are mixed and pulverized to obtain a traditional Chinese medicine compound powder. The traditional Chinese medicine compound powder is then fermented with a mixed probiotic composed of Trichoderma reesei, Fussella fussula, and Aspergillus niger.

[0013] Preferably, the volume ratio of Trichoderma reesei, Fusionia fussima and Aspergillus niger is 1-5:1-3:1, and more preferably 1:1:1.

[0014] Furthermore, the fermentation method includes the following steps:

[0015] ① Take the compound powder of traditional Chinese medicine and add distilled water at a mass ratio of 1:1 to make the medicine reach a water content of 50%, which is conducive to the fermentation of probiotics. After stirring evenly, add brown sugar, yeast extract and CaCO3, and make up to 1000ml. The final concentration of brown sugar is 20g / L, the final concentration of yeast extract is 4g / L, and the final mass concentration of CaCO3 is 1.5%. Adjust the pH value to 7.0, dispense into 250ml wide-mouth Erlenmeyer flasks, put them in an autoclave and keep the temperature at 121℃ for 15min. After sterilization, take them out and let them cool naturally to room temperature to obtain the medicinal material fermentation culture medium.

[0016] ② Inoculate Trichoderma reesei, Fussula fusionis and Aspergillus niger liquids respectively. After inoculation, place them in a constant temperature and humidity incubator. Under the conditions of 28℃ and 70% relative humidity, first ferment aerobically for 24 hours and then anaerobically for 24 hours. After fermentation, dry the fermented Chinese medicine compound at 50℃ for later use.

[0017] The inoculation amounts of Trichoderma reesei, Fusionia fussima and Aspergillus niger are 5-10% of the mass of the medicinal material fermentation culture medium, preferably 8%.

[0018] Fourthly, the present invention provides the use of the traditional Chinese medicine compositions described in the first and second aspects above in the preparation of drugs for the prevention and / or treatment of bacterial pneumonia in pigs.

[0019] Furthermore, the porcine bacterial pneumonia includes Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis.

[0020] Fifthly, the present invention provides a medicament for the prevention and / or treatment of bacterial pneumonia in pigs, comprising the traditional Chinese medicine composition described in the first and second aspects above and medically acceptable excipients; the dosage form of the medicament is any pharmaceutically acceptable dosage form, which is any one of sugar-coated tablets, capsules, granules, syrups, and pills.

[0021] The advantages and beneficial effects of this invention are:

[0022] This invention combines traditional and modern preparation techniques to research and clinically apply traditional Chinese medicine. It selects a compound traditional Chinese medicine formula prepared by co-fermentation of three probiotics: *Trichoderma reesei*, *Westernella fusionis*, and *Aspergillus niger*. This formula addresses the aforementioned problems in the prevention and treatment of bacterial pneumonia in pigs. Clinical application shows that the compound formula of this invention achieves a cure rate of 91.83% for bacterial pneumonia in pigs. After co-fermentation with *Trichoderma reesei*, *Westernella fusionis*, and *Aspergillus niger*, the cure rate reaches 95.97%, improving efficacy, reducing recurrence, and eliminating drug residues. Furthermore, all components are common traditional Chinese medicines, resulting in low cost and high safety. This approach reduces economic losses for farmers and meets the demand for green food, aligning with development trends.

[0023] Traditional Chinese medicine (TCM) classifies pneumonia as belonging to the categories of lung heat, cough, and wind-heat, caused by the invasion of heat toxins that affect the lung's defensive qi, leading to impaired lung function. The Ma Xing Shi Gan Tang formula from the *Shang Han Lun* (Treatise on Cold Damage) has the effects of clearing lung heat and promoting lung function. This invention is a modified version of Ma Xing Shi Gan Tang. The principal ingredient is gypsum, which is pungent, sweet, and very cold, entering the lung and stomach meridians. It has the effect of dispersing stagnant lung heat and is the main antipyretic. Ephedra is also a principal ingredient, pungent and slightly bitter, entering the lung and bladder meridians, and has the effect of inducing sweating. Combined with gypsum, it helps to expel pathogens and relieve asthma. The assistant ingredient is apricot kernel, bitter and slightly warm, entering the lung and large intestine meridians. It tends to nourish lung qi, assisting ephedra in relieving asthma and gypsum in clearing lung heat. The adjuvant ingredient is licorice root, which harmonizes the medicinal properties and alleviates asthma. Gardenia clears heat and drains fire, cools the blood and detoxifies. Honeysuckle and forsythia clear heat and detoxify. Astragalus membranaceus is bitter and cold in nature, and enters the lung and spleen meridians. It has the effects of clearing heat and drying dampness, purging fire and detoxifying, invigorating qi and strengthening the spleen, and promoting diuresis and reducing swelling. Molecular biology suggests that astragalus membranaceus can activate lymphocyte function, increase lymphocyte transformation rate, and enhance immunity. Rhubarb has the effects of clearing damp-heat, purging fire, cooling blood, removing blood stasis, and detoxifying. Glauber's salt has the effects of clearing heat and reducing swelling, and purging. Peucedanum praeruptorum disperses wind-heat, lowers qi and resolves phlegm. Pueraria lobata relieves exterior symptoms and reduces fever, and promotes body fluid production. The addition of honeysuckle, gypsum, and forsythia in this formula enhances the effects of clearing qi and cooling blood, and promoting qi circulation, significantly improving the effects of clearing heat and detoxifying. Talc combined with astragalus membranaceus has the effects of opening the orifices and clearing heat and promoting diuresis, significantly improving the body's immunity. This invention has a scientific formulation, with synergistic effects among the components, exhibiting significant antibacterial and anti-inflammatory effects, and without drug resistance.

[0024] This study combines modern microbial engineering technology with traditional Chinese medicine processing techniques, selecting *Trichoderma reesei*, *Westernella fusionis*, and *Aspergillus niger* to ferment traditional Chinese medicine. These three probiotics synergistically act on the components of the medicine, enabling a more complete release of active ingredients and thus improving drug activity. The synergistic effect of these three probiotics—*Trichoderma reesei*, *Westernella fusionis*, and *Aspergillus niger*—involves the secretion of large amounts of decomposing enzymes and extracellular enzymes, which can further disrupt the cell walls of the medicinal materials, promoting the dissolution of active ingredients and thus enhancing efficacy. The fermented product contains a large number of active enzymes that can be absorbed by the body to regulate immune function and enhance antibacterial ability. Compounds with antibacterial activity, such as bacteriocins, lactic acid, and glucan, are generated during fermentation, directly contributing to improved efficacy. The *Westernella fusionis* selected in this invention has advantages such as resistance to gastric acid and bile salts, and strong adhesion to intestinal epithelial cells, directly affecting the regulation of the intestinal flora-neuroendocrine-immune axis. The synergistic effect of three probiotics—Trichoderma reesei, Weissella fusionis, and Aspergillus niger—further enhances their direct efficacy in regulating the gut microbiota-neuroendocrine-immune axis. Fermentation processing can reduce the side effects of the drugs. In this formula, rhubarb, ephedra, apricot kernel, gypsum, astragalus, anemarrhena, gardenia, talc, licorice, honeysuckle, forsythia, mirabilite, angelica, and kudzu root, after fermentation, results in a decrease in total and bound anthraquinone content and an increase in free anthraquinone content, which can mitigate gastrointestinal irritation and reduce laxative effects. In this invention, Trichoderma reesei, Weissella fusionis, and Aspergillus niger are inoculated separately and placed in a constant temperature and humidity incubator. After aerobic fermentation for 24 hours at 28°C and 70% relative humidity, followed by anaerobic fermentation for another 24 hours, this step allows the three bacteria to grow synergistically and secrete more effective enzymes beneficial for fermenting traditional Chinese medicine. Meanwhile, traditionally processed Chinese medicines are in the form of aqueous solutions or powders, possessing a pungent odor. Pigs have sensitive senses of smell and taste, and mixing bitter-tasting medicines into their feed will reduce feed intake, thus decreasing efficacy. Furthermore, a strong odor makes it difficult for animals to willingly consume the medicine. This invention can prepare sugar-coated tablets or pills, which, when mixed into feed, effectively solves this problem. Moreover, pigs are herd animals, and in intensive production, administering medication to each pig individually is very labor-intensive. The Chinese medicines in this invention, fermented with *Trichoderma reesei*, *Weissella fusionis*, and *Aspergillus niger*, possess a unique fermented aroma, greatly improving palatability. Detailed Implementation

[0025] The technical solutions in the embodiments of this application are clearly described below. Obviously, the described embodiments are only some, not all, of the embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments of this application are within the scope of protection of this application.

[0026] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0027] Example 1

[0028] This embodiment provides a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components:

[0029] Ephedra 40g, Apricot kernel 10g, Gypsum 80g, Astragalus 40g, Anemarrhena 40g, Gardenia 40g, Talc 30g, Licorice 10g, Honeysuckle 30g, Forsythia 10g, Rhubarb 10g, Glauber's salt 10g, Peucedanum praeruptorum 30g, Kudzu root 40g.

[0030] After mixing the above ingredients, pulverize them and pass them through a 10-mesh sieve to obtain a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs.

[0031] Example 2

[0032] This embodiment provides a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components:

[0033] Ephedra 50g, Apricot kernel 20g, Gypsum 100g, Astragalus 50g, Anemarrhena 50g, Gardenia 50g, Talc 40g, Licorice 10g, Honeysuckle 40g, Forsythia 10g, Rhubarb 10g, Glauber's salt 10g, Peucedanum praeruptorum 40g, Kudzu root 50g.

[0034] After mixing the above ingredients, pulverize them and pass them through a 10-mesh sieve to obtain a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs.

[0035] Example 3

[0036] This embodiment provides a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components:

[0037] Ephedra 60g, Apricot kernel 20g, Gypsum 120g, Astragalus 60g, Anemarrhena 60g, Gardenia 60g, Talc 50g, Licorice 20g, Honeysuckle 50g, Forsythia 20g, Rhubarb 15g, Glauber's salt 15g, Peucedanum praeruptorum 50g, Kudzu root 60g.

[0038] After mixing the above ingredients, pulverize them and pass them through a 10-mesh sieve to obtain a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs.

[0039] Example 4

[0040] This embodiment provides a traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, comprising the following components:

[0041] Ephedra 60g, Apricot kernel 20g, Gypsum 120g, Astragalus 60g, Anemarrhena 60g, Gardenia 60g, Talc 50g, Licorice 20g, Honeysuckle 50g, Forsythia 20g, Rhubarb 15g, Glauber's salt 15g, Peucedanum praeruptorum 50g, Kudzu root 60g.

[0042] The method for preparing the traditional Chinese medicine composition in this example is as follows:

[0043] After mixing the above ingredients, pulverize them and pass them through a 10-mesh sieve to obtain a traditional Chinese medicine compound powder, which is then fermented. The fermentation method includes the following steps:

[0044] (1) Preparation of medicinal material fermentation culture medium: Take the compound powder of Chinese medicine, pass it through a 10-mesh sieve, weigh it, add distilled water in a mass ratio of 1:1, stir evenly, add brown sugar, yeast extract and CaCO3 to make up to 1000ml, the final concentration of brown sugar is 20g / L, the final concentration of yeast extract is 4g / L, the final mass concentration of CaCO3 is 1.5%, adjust the pH value to 7.0, stir evenly, dispense into 250ml wide-mouth triangular flasks, put them in a high-pressure steam sterilizer and keep the temperature at 121℃, sterilize for 15 min, take them out and let them cool naturally to room temperature.

[0045] (2) Microbial strains

[0046] Trichoderma reesei, Weissella fusionis, and Aspergillus niger were all provided and preserved by the laboratory of the College of Veterinary Medicine, Sichuan Agricultural University, and stored on MRS solid medium slant at 4°C. All three bacteria were revived by seed culture after being preserved on MRS solid medium slant.

[0047] MRS solid culture medium: 10g protein powder, 5g beef meal, 4g yeast powder, 2g glucose, 1ml Tween 80, 2g dipotassium hydrogen phosphate, 5g sodium acetate, 2g triammonium citrate, 0.2g magnesium sulfate, 0.05g manganese sulfate, 15g agar powder, 1000ml distilled water; natural pH, sterilize at 121℃ for 20 min.

[0048] Liquid seed culture medium: glucose 20 g / L, peptone 10 g / L, yeast extract 10 g / L, KH2PO4 1 g / L, MgSO4 0.5 g / L, natural pH, sterilized at 121℃ for 20 min.

[0049] (3) Preparation of starter culture of Trichoderma reesei, Fusionia fuscin and Aspergillus niger:

[0050] Take the medicinal fermentation culture medium prepared in step (1), and inoculate Trichoderma reesei, Fusionia fuscin and Aspergillus niger liquid in a biosafety cabinet. The inoculation amount of Trichoderma reesei, Fusionia fuscin and Aspergillus niger liquid is 8% of the mass of the medicinal fermentation culture medium. After inoculation, place it in a constant temperature and humidity incubator. Under the conditions of 28℃ and 70% relative humidity, first ferment aerobic culture for 24 hours and then anaerobic culture for 24 hours. After the fermentation is completed, dry the fermented Chinese medicine composition at 50℃ for later use.

[0051] Comparative Example 1

[0052] This comparative example provides a traditional Chinese medicine composition for the prevention and treatment of bacterial pneumonia in pigs. The only difference between this comparative example and Example 3 is that Plantago asiatica is used instead of talc, and Isatis indigotica is used instead of honeysuckle. The rest is the same as Example 3.

[0053] Comparative Example 2

[0054] This comparative example provides a traditional Chinese medicine composition for the prevention and treatment of bacterial pneumonia in pigs. The only difference between this comparative example and Example 3 is that the amount of talc is 20g and the amount of honeysuckle is 20g, and the rest is the same as Example 3.

[0055] Experimental Example

[0056] A treatment trial

[0057] (a) Treatment drugs

[0058] The test drugs were the traditional Chinese medicine compositions prepared in Examples 1-3 and Comparative Examples 1-2, the fermented traditional Chinese medicine composition prepared in Example 4, and the Western medicine group.

[0059] Among them, the traditional Chinese medicine compositions prepared in Examples 1-3 and Comparative Examples 1 and 2 need to be prepared into traditional Chinese medicine extracts. The extraction method is as follows: the traditional Chinese medicine is extracted three times by conventional water decoction, each time for 30 minutes. The liquids after the three decoctions are combined, centrifuged at 4000 r / min for 15 minutes, and then cooled to obtain the traditional Chinese medicine extract.

[0060] The Western medicine group was treated with tilmicosin.

[0061] (II) Treatment methods

[0062] ① Experimental animals

[0063] From April 2021 to August 2023, the experiment was conducted at a moderately sized pig farm in Suining City. After natural onset, the pigs were diagnosed clinically and laboratory-wise as being infected with *Actinomyces pleuropneumoniae*, *Pasteurella multocida*, and *Haemophilus parasuis*. The healthy control group and the preventative experimental group also came from the same farm. All pigs were three-way crossbred pigs. The experimental pigs were conventionally fed, and their immunization program followed the normal procedures of the pig farm. They had free access to water and feed. The purpose of this experiment was to study the clinical efficacy of traditional Chinese medicine compositions, fermented traditional Chinese medicine compositions, and common Western medicines in the prevention and treatment of bacterial pneumonia.

[0064] ② Treatment group

[0065] This experiment consisted of 7 groups: Comparative Example 1, Comparative Example 2, Example 1, Example 2, Example 3, Example 4, and Western medicine group. Each group had 3 replicates. Each group also had 1 drug control group, 3 positive control groups, and 1 negative control group. The experiment was conducted independently at 98 sites, with 30 animals at each site, for a total of 2940 animals.

[0066] Example 1 contains 14 groups of traditional Chinese medicine extracts, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine extracts from Example 1, including three groups each for Actinobacillus pleuropneumoniae treatment, Pasteurella multocida treatment, and Haemophilus parasuis treatment.

[0067] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 42-45 days, the second group had an age of onset of 81-85 days, and the third group had an age of onset of 99-102 days.

[0068] In the Pasteurella multocida treatment group, the age of onset was 72-76 days in the first group, 110-115 days in the second group, and 103-108 days in the third group.

[0069] In the Haemophilus parasuis treatment group, the first group had an onset age of 40-45 days, the second group had an onset age of 32-35 days, and the third group had an onset age of 43-49 days.

[0070] The age of onset in the Actinobacillus pleuropneumoniae treatment group and the positive control group was 82-86 days.

[0071] The age of onset in the positive control group of the Pasteurella multocida treatment group was 112-117 days.

[0072] The age of onset in the positive control group of the Haemophilus parasuis treatment group was 45-49 days.

[0073] Example 2 included 14 groups of traditional Chinese medicine extracts, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine extracts from Example 2, including three groups each of Actinobacillus pleuropneumoniae treatment, Pasteurella multocida treatment, and Haemophilus parasuis treatment.

[0074] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 75-79 days, the second group had an age of onset of 108-112 days, and the third group had an age of onset of 89-92 days.

[0075] In the Pasteurella multocida treatment group, the age of onset was 65-70 days in the first group, 72-76 days in the second group, and 103-108 days in the third group.

[0076] In the Haemophilus parasuis treatment group, the first group had an onset age of 35-41 days, the second group had an onset age of 43-48 days, and the third group had an onset age of 85-90 days.

[0077] The age of onset in the Actinobacillus pleuropneumoniae treatment group and the positive control group was 75-81 days.

[0078] The age of onset in the positive control group of the Pasteurella multocida treatment group was 89-94 days.

[0079] The age of onset in the positive control group of the Haemophilus parasuis treatment group was 32-36 days.

[0080] Example 3 included 14 groups of traditional Chinese medicine extracts, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine extracts from Example 3, including three groups each of Actinobacillus pleuropneumoniae treatment, Pasteurella multocida treatment, and Haemophilus parasuis treatment.

[0081] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 86-91 days, the second group had an age of onset of 99-102 days, and the third group had an age of onset of 74-80 days.

[0082] In the Pasteurella multocida treatment group, the age of onset was 75-81 days in the first group, 110-115 days in the second group, and 81-89 days in the third group.

[0083] In the Haemophilus parasuis treatment group, the first group had an onset age of 35-41 days, the second group had an onset age of 32-36 days, and the third group had an onset age of 43-49 days.

[0084] The age of onset in the Actinobacillus pleuropneumoniae treatment group and the positive control group was 74-79 days.

[0085] The age of onset in the positive control group of the Pasteurella multocida treatment group was 83-86 days.

[0086] The age of onset in the Haemophilus parasuis treatment group and the positive control group was 38-43 days.

[0087] Example 4: The traditional Chinese medicine fermentation group consisted of 14 groups, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine fermentation composition of Example 4, including three groups each of Actinobacillus pleuropneumoniae treatment, Pasteurella multocida treatment, and Haemophilus parasuis treatment.

[0088] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 89-82 days, the second group had an age of onset of 84-88 days, and the third group had an age of onset of 76-79 days.

[0089] In the Pasteurella multocida treatment group, the age of onset was 88-92 days in the first group, 94-98 days in the second group, and 81-86 days in the third group.

[0090] In the Haemophilus parasuis treatment group, the first group had an onset age of 56-59 days, the second group had an onset age of 34-39 days, and the third group had an onset age of 42-47 days.

[0091] The age of onset in the Actinobacillus pleuropneumoniae treatment group and the positive control group was 85-99 days.

[0092] The age of onset in the positive control group of the Pasteurella multocida treatment group was 91-96 days.

[0093] The age of onset of the disease in the Haemophilus parasuis treatment group and the positive control group was 25-30 days.

[0094] Comparative Example 1 consisted of 14 groups of traditional Chinese medicine extracts, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine extracts from Comparative Example 1, including three groups each of the Actinobacillus pleuropneumoniae treatment group, Pasteurella multocida treatment group, and Haemophilus parasuis treatment group.

[0095] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 112-116 days, the second group had an age of onset of 83-89 days, and the third group had an age of onset of 86-91 days.

[0096] In the Pasteurella multocida treatment group, the age of onset was 85-92 days in the first group, 75-81 days in the second group, and 110-115 days in the third group.

[0097] In the Haemophilus parasuis treatment group, the first group had an onset age of 32-36 days, the second group had an onset age of 43-49 days, and the third group had an onset age of 51-56 days.

[0098] The age of onset in the Actinobacillus pleuropneumoniae treatment group and the positive control group was 89-91 days.

[0099] The age of onset in the positive control group of the Pasteurella multocida treatment group was 91-96 days.

[0100] The age of onset of the disease in the Haemophilus parasuis treatment group and the positive control group was 25-30 days.

[0101] Comparative Example 2 consisted of 14 groups of traditional Chinese medicine extracts, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine experimental groups. The nine experimental groups were treated with the traditional Chinese medicine extracts from Comparative Example 2, including three groups each of the Actinobacillus pleuropneumoniae treatment group, Pasteurella multocida treatment group, and Haemophilus parasuis treatment group.

[0102] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 69-73 days, the second group had an age of onset of 102-108 days, and the third group had an age of onset of 76-81 days.

[0103] In the Pasteurella multocida treatment group, the age of onset was 81-89 days in the first group, 112-118 days in the second group, and 75-82 days in the third group.

[0104] In the Haemophilus parasuis treatment group, the first group had an onset age of 31-34 days, the second group had an onset age of 42-47 days, and the third group had an onset age of 51-56 days.

[0105] The age of onset in the positive control group of the Actinobacillus pleuropneumoniae treatment group was 102-108 days.

[0106] The age of onset in the positive control group of the Pasteurella multocida treatment group was 75-82 days.

[0107] The age of onset in the positive control group of the Haemophilus parasuis treatment group was 51-56 days.

[0108] The Western medicine treatment group consisted of 14 groups, including one positive control group for Actinobacillus pleuropneumoniae treatment, one positive control group for Pasteurella multocida treatment, one positive control group for Haemophilus parasuis treatment, one drug control group, one healthy control group, and nine Western medicine groups. The nine Western medicine groups were treated with tilmicosin, including three groups each for Actinobacillus pleuropneumoniae treatment, Pasteurella multocida treatment, and Haemophilus parasuis treatment.

[0109] Among the Actinobacillus pleuropneumoniae treatment groups, the first group had an age of onset of 73-76 days, the second group had an age of onset of 79-84 days, and the third group had an age of onset of 96-100 days.

[0110] In the Pasteurella multocida treatment group, the age of onset was 85-89 days in the first group, 108-113 days in the second group, and 76-81 days in the third group.

[0111] In the Haemophilus parasuis treatment group, the first group had an onset age of 37-42 days, the second group had an onset age of 37-42 days, and the third group had an onset age of 29-34 days.

[0112] The age of onset in the positive control group of the Actinobacillus pleuropneumoniae treatment group was 78-84 days.

[0113] The age of onset in the positive control group of the Pasteurella multocida treatment group was 96-102 days.

[0114] The age of onset in the Haemophilus parasuis treatment group and the positive control group was 26-31 days.

[0115] ③ Treatment methods

[0116] At the site of the outbreak, strict feeding management and disinfection should be implemented as required.

[0117] The dosage and administration for comparative examples 1-2 and examples 1-3 in treating Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis were as follows: 200 ml / head / time for nursery pigs, once a day; 300 ml / head / time for fattening pigs, once a day, administered via drinking water (oral administration is required during the acute infection period). One course of treatment lasted 5-7 days, and observation continued for 5-7 days after discontinuation of the medication.

[0118] Example 4: The dosage for treating Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis is 0.3-0.5 g / kg body weight per head, once a day, orally, for 5-7 days as one course of treatment. After stopping the medication, continue to observe for 5-7 days.

[0119] Western medicine treatment group: Tilmicosin was used for treatment. 20% tilmicosin premix was selected, mixed into feed, 1000-2000g / 1000kg, for 15 consecutive days.

[0120] All experimental groups were clinically cured when clinical symptoms disappeared and water and feed intake returned to normal. The effectiveness was further confirmed by combining the results of pathological necropsy and random sampling. Indicators such as the prevention and treatment effectiveness rate were statistically analyzed. Production levels were observed, and the mental state, appetite, feces, and mortality of pigs in each group were recorded. For pigs that died in each group, pathological necropsy, isolation culture, and biochemical identification were performed to determine whether the cause of death was Actinobacillus pleuropneumoniae, Pasteurella multocida, Haemophilus parasuis, or the drug itself. A comprehensive evaluation was conducted to determine the overall effectiveness of the compound preparation and the drug preparation process. During the experiment, the feeding and management conditions of all groups were consistent and essentially the same as before the experiment.

[0121] Second prevention trial

[0122] (a) Preventive drugs

[0123] The test drugs were the fermented traditional Chinese medicine composition prepared in Example 4 and the Western medicine group.

[0124] The Western medicine group used tilmicosin for prevention.

[0125] (II) Prevention methods

[0126] ①Prevention of animals and grouping

[0127] The prevention group consisted of 9 farms (not belonging to the same herd) where bacterial pneumonia cases had occurred. Each farm was divided into 18 groups of 60 pigs, housed separately in individual pens. There were 6 groups each for Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis. For each disease, the 6 groups received preventative treatment using either the fermented traditional Chinese medicine composition prepared in Example 4 or Western medicine mixed with feed. The Western medicine prevention group received the same routine preventative medication.

[0128] ②Prevention methods

[0129] Actinobacillus pleuropneumoniae prevention test group: In Example 4 group, the drug was administered at 0.1 g / kg body weight per head, mixed with feed, once a day, at 35-40 days and 70-75 days of age, respectively, for 5 days each time. The western medicine group received western medicine within the same time period. Observation continued from the start of medication until 150 days of age, and the incidence and mortality were recorded.

[0130] Haemophilus parasuis prevention experimental group: In Example 4 group, the dosage was 0.1 g / kg body weight per head, mixed with feed, once a day, with additional medication at 15-20 days, 35-40 days, and 70-75 days of age, each time for 5 days. The western medicine group received western medicine during the same time period. Continuous observation was carried out from the start of medication until 150 days of age, and the incidence and mortality were recorded.

[0131] Pasteurella multocida prevention test group: During the hot and humid season (May-September), experimental pigs were treated with the medication described in Example 4 at a rate of 0.1 g / kg body weight per head, mixed with feed, once a day, every 10 days, for 3 days each time. The Western medicine group received the Western medicine for the same period. Observations were conducted continuously from the start of medication until 150 days of age, and morbidity and mortality were recorded.

[0132] During the experiment, the feeding and management conditions of all groups were consistent and basically the same as before the experiment. Pathological examination, isolation culture, and biochemical identification of the dead pigs confirmed that they were caused by Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis.

[0133] Results of the three experiments

[0134] (1) Criteria for judging therapeutic efficacy

[0135] During the mortality test, pigs that exhibited typical symptoms and died, showed characteristic lesions upon necropsy, and had pathogens isolated from relevant tissues were defined as infected and died. The percentage of infected pigs that died out of the total number of pigs in each group is the mortality rate for that group.

[0136] Effective treatment was defined as the partial disappearance of clinical symptoms after medication and the basic return to normal of physiological indicators such as appetite, water intake, and mental state. The effective rate for each group was calculated based on the number of effective cases.

[0137] The cure rate is defined as the complete disappearance of clinical symptoms after medication, and the complete return to normal of physiological indicators such as appetite, water intake, and mental state. The percentage of cured cases out of the total number of cases in this trial group is the cure rate.

[0138] Ineffectiveness is defined as no relief of clinical symptoms, or even worsening or death. The ineffectiveness rate of each group is calculated based on the number of ineffective cases.

[0139] (2) Statistical analysis

[0140] Experimental data were analyzed using SPSS 20.0 software, and multiple comparisons were performed using Duncan's method. Results are expressed as mean ± standard deviation, and p < 0.05 was considered statistically significant.

[0141] (3) Experimental results

[0142] 3.1 Results of the treatment trial

[0143] Clinical observations revealed no abnormalities in the negative controls and drug control groups of each comparative example and embodiment. The positive control group exhibited obvious clinical symptoms.

[0144] As shown in Table 1, the results of the treatment trial indicate that both the Example 4 group and the Western medicine group have good therapeutic effects on swine pneumonia caused by the three pathogenic bacteria.

[0145] As shown in Tables 1-3, in the Actinobacillus pleuropneumoniae treatment trial, the cure rate and efficacy of the fermented traditional Chinese medicine composition in Example 4 were significantly different from those of Western medicine.

[0146] In the efficacy trial of Pasteurella multocida treatment, the efficacy of the fermented traditional Chinese medicine composition in Example 4 was significantly different from that of Western medicine.

[0147] In the trial of the therapeutic effect of Haemophilus parasuis, the cure rate and efficacy of the fermented traditional Chinese medicine composition in Example 4 were significantly different from those of Western medicine.

[0148] Table 1. Statistical analysis results of the treatment effects in each experimental case.

[0149]

[0150] Table 2: Results of T-test analysis of cure rate for each experimental case

[0151]

[0152] Note: P ≥ 0.05 indicates no significant difference; P < 0.05 indicates a significant difference; P ≤ 0.01 indicates an extremely significant difference.

[0153] 3.2 Prevention Results

[0154] As shown in Table 3, the results of the prevention test indicate that fermented traditional Chinese medicine and Western medicine have good preventive effects against swine pneumonia caused by three pathogenic bacteria, and the preventive effect of fermented traditional Chinese medicine is better than that of Western medicine.

[0155] Table 3. Results of Example 4 and Western medicine in preventing three types of bacterial pneumonia in pigs (three prevention points)

[0156] Unit: heads, %

[0157]

[0158] Note: During the trial in each natural population, the Western medicine group used the same medication as the daily preventive medication.

[0159] Table 4. Statistical Analysis of Example 4 and the Western Medicine Group in the Prevention Experiment

[0160]

[0161] Note: P ≥ 0.05 indicates no significant difference; P < 0.05 indicates a significant difference; P ≤ 0.01 indicates an extremely significant difference.

[0162] As can be seen from Table 3-4:

[0163] Example 4 demonstrates a significant preventative effect against Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis in pigs, with a preventative effect far superior to that of Western medicine. It is both superior and reliable.

[0164] The embodiments of this application have been described above, but this application is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of this application without departing from the spirit and scope of the claims, and all of these forms are within the protection scope of this application.

Claims

1. A traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs, wherein the bacterial pneumonia is caused by at least one of Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis, characterized in that, It is made from the following ingredients by weight: 40 parts ephedra, 20 parts apricot kernel, 120 parts gypsum, 60 parts astragalus, 60 parts anemarrhena, 60 parts gardenia, 50 parts talc, 20 parts licorice, 50 parts honeysuckle, 20 parts forsythia, 15 parts rhubarb, 15 parts mirabilite, 50 parts angelica, and 60 parts kudzu root; and is prepared by fermentation. The preparation method of the traditional Chinese medicine composition includes the following steps: Ephedra, apricot kernel, gypsum, astragalus, anemarrhena, gardenia, talc, licorice, honeysuckle, forsythia, rhubarb, mirabilite, angelica and kudzu root are mixed and pulverized to obtain a traditional Chinese medicine compound powder. The traditional Chinese medicine compound powder is then fermented with a mixed probiotic composed of Trichoderma reesei, Fussella fusionis and Aspergillus niger. The volume ratio of Trichoderma reesei, Fusion Weissella and Aspergillus niger is 1:1:1; The fermentation method includes the following steps: ① Take the compound powder of traditional Chinese medicine, add distilled water at a mass ratio of 1:1, stir well, then add brown sugar, yeast extract and CaCO3 to make up to 1000ml. The final concentration of brown sugar is 20g / L, the final concentration of yeast extract is 4g / L, and the final mass concentration of CaCO3 is 1.5%. Adjust the pH value to 7.0, dispense into 250ml wide-mouth triangular flasks, put them in a high-pressure steam sterilizer and keep the temperature at 121℃. After sterilization for 15min, take them out and let them cool naturally to room temperature to obtain the medicinal material fermentation culture medium. ② Inoculate Trichoderma reesei, Fussula fusionis and Aspergillus niger liquids respectively. After inoculation, place them in a constant temperature and humidity incubator. Under the conditions of 28℃ and 70% relative humidity, first ferment aerobic culture for 24 hours and then anaerobic culture for 24 hours. After fermentation, dry the fermented Chinese medicine compound at 50℃ for later use. The inoculation amounts of Trichoderma reesei, Fusionia fussima, and Aspergillus niger were 5-10% of the mass of the medicinal material fermentation medium.

2. The traditional Chinese medicine composition for preventing and treating bacterial pneumonia in pigs according to claim 1, characterized in that, The inoculation amounts of Trichoderma reesei, Fusionia fussima, and Aspergillus niger were 8% of the mass of the medicinal material fermentation medium.

3. The use of the traditional Chinese medicine composition according to claim 1 or 2 in the preparation of a drug for the prevention and / or treatment of porcine bacterial pneumonia, wherein the porcine bacterial pneumonia is caused by at least one of Actinobacillus pleuropneumoniae, Pasteurella multocida, and Haemophilus parasuis.

4. A drug for the prevention and / or treatment of bacterial pneumonia in pigs, characterized in that, The drug is made from the traditional Chinese medicine composition as described in claim 1 or 2 and medically acceptable excipients; the dosage form of the drug is any one of sugar-coated tablets, capsules, granules, syrups, and pills.

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