A traditional Chinese medicine composition for treating staphylococcus psittaci infection and a preparation method thereof

Abstract

The application provides a traditional Chinese medicine composition for treating S. psittaci infection and a preparation method thereof, and relates to the technical field of veterinary traditional Chinese medicine preparation.The traditional Chinese medicine composition is prepared by decocting and extracting honeysuckle, forsythia, radix scrophulariae, rhizoma coptidis, dandelion, wild chrysanthemum, purple flowered ground ivy and licorice, and then concentrating the filtrate.The application is based on the theory of traditional Chinese veterinary medicine, and follows the treatment principle of "clearing heat and resolving toxins, eliminating swelling and pus, cooling blood and removing stasis, and strengthening body resistance and eliminating pathogenic factors".The drugs are scientifically matched.The in-vitro bacteriostatic test shows that the composition has obvious inhibitory and killing effects on S. aureus, the minimum inhibitory concentration (MIC) is 0.5 mg / mL, and the minimum bactericidal concentration (MBC) is 1.0 mg / mL.The composition is significantly better than the existing patent technology composition.The clinical treatment test shows that the composition has a cure rate of 85.0% for S. psittaci infection, and a total effective rate of 95.0%, which is better than the curative effect of doxycycline hydrochloride, can replace part of the antibiotics, has no toxic side effects, and is high in safety.

Description

Technical Field

[0001] This invention relates to the field of veterinary traditional Chinese medicine, specifically a traditional Chinese medicine composition for treating Staphylococcus psittaci infection and its preparation method. Background Technology

[0002] Staphylococcus aureus is a bacterial disease caused by Staphylococcus aureus, which is an acute or chronic bacterial illness affecting poultry, mammals, and humans. It is characterized by localized suppuration of tendon sheaths, joints, and synovial bursae, wound infections, septicemia, and omphalitis. In pet birds such as parrots, Staphylococcus aureus infection can manifest in various forms, including acute septicemia, arthritis, wingtip gangrene, and ocular infections. Ocular cases are also common and can lead to death in severe cases.

[0003] Currently, antibiotic therapy, such as tetracyclines, gentamicin, and levofloxacin, is the main clinical treatment for Staphylococcus aureus infections. However, the problem of drug resistance in Staphylococcus aureus is becoming increasingly serious. Studies have shown that Staphylococcus aureus has developed varying degrees of resistance to many commonly used antibiotics, and the resistance spectrum is constantly expanding. In recent years, although some traditional Chinese medicine combinations have been used to treat avian staphylococcal infections, many technical limitations still exist.

[0004] For example, Chinese patent CN201811627013.6 (Traditional Chinese Medicine Composition for Treating Staphylococcal Infection in Chickens and its Preparation Method) discloses a traditional Chinese medicine composition for treating staphylococcal infection in chickens. Its components include Isatis root, Cyperus rhizome, Magnolia bark, Plantago seed, Dandelion, Scutellaria root, Cirsium root, Tangerine peel, and Rhubarb, possessing antibacterial, anti-inflammatory, heat-clearing, and dampness-removing effects. However, this composition is designed specifically for staphylococcal infection in chickens and is not a formula specifically for parrots. The preparation process lacks clarity in its specific implementation details. This patent does not provide pharmacodynamic experimental data to prove its antibacterial effect, nor does it conduct optimization research on the compatibility and synergistic effects, lacking core technological support for its inventiveness. Furthermore, this composition mainly focuses on "clearing heat and removing dampness," failing to fully cover the complete pathogenesis of staphylococcal infection, such as "heat toxicity accumulation, blood stasis, sores, swelling, and pain."

[0005] For example, Chinese patent CN202011610006.2 (Traditional Chinese Medicine Composition for Treating Staphylococcal Infection in Chickens and its Preparation Process) discloses a traditional Chinese medicine composition whose components include stir-fried Atractylodes macrocephala, stir-fried Dioscorea opposita, Halloysite, poppy shell, and Plantago asiatica, which has antibacterial, anti-inflammatory, heat-clearing, and detoxifying effects. This composition mainly focuses on strengthening the spleen and astringing to stop diarrhea, and does not fully cover the main pathological aspect of Staphylococcus aureus infection—namely, the core treatment links of detoxification, pus drainage, swelling reduction, and nodule dissipation. At the same time, its preparation process is relatively simple and does not optimize the extraction efficiency of the effective components of traditional Chinese medicine, making it difficult to ensure the full release and stability of the efficacy. Although this patent shows "inhibition of the growth of Staphylococcus aureus," it also lacks detailed in vitro antibacterial data and clinical efficacy data as support.

[0006] Furthermore, as pet birds, parrots exhibit significant physiological differences from domestic poultry. Parrots are smaller, have a higher metabolic rate, and their sensitivity and tolerance to drugs are not entirely the same as those of domestic chickens. When existing traditional Chinese medicine compositions designed for domestic chickens are directly applied to parrots, problems such as inaccurate dosage and unstable efficacy may arise.

[0007] Therefore, those skilled in the art provide a traditional Chinese medicine composition for treating Staphylococcus psittaci infection and its preparation method to solve the problems mentioned in the background art. Summary of the Invention

[0008] The purpose of this invention is to provide a traditional Chinese medicine composition for treating Staphylococcus psittaci infection and its preparation method, so as to solve the problems mentioned in the background art.

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] A traditional Chinese medicine composition for treating Staphylococcus psittaci infection, made from the following raw materials in parts by weight:

[0011] Honeysuckle 10-20 parts; Forsythia 8-16 parts; Scutellaria baicalensis 6-12 parts; Coptis chinensis 4-8 parts; Taraxacum mongolicum 10-18 parts; Chrysanthemum indicum 6-12 parts; Viola yedoensis 8-14 parts; Glycyrrhiza uralensis 3-8 parts.

[0012] Preferably, the composition is made from the following parts by weight of active pharmaceutical ingredient:

[0013] Honeysuckle 12-18 parts; Forsythia 10-14 parts; Scutellaria baicalensis 8-10 parts; Coptis chinensis 5-7 parts; Taraxacum mongolicum 12-16 parts; Chrysanthemum indicum 8-10 parts; Viola yedoensis 10-12 parts; Glycyrrhiza uralensis 4-6 parts.

[0014] As the most preferred embodiment, the composition is made from the following parts by weight of active pharmaceutical ingredient:

[0015] Honeysuckle 15 parts; Forsythia 12 parts; Scutellaria baicalensis 9 parts; Coptis chinensis 6 parts; Dandelion 15 parts; Chrysanthemum indicum 9 parts; Viola yedoensis 12 parts; Licorice 6 parts.

[0016] Honeysuckle: It is cold in nature and sweet in taste. It enters the lung, heart and stomach meridians. Its main function is to clear heat and detoxify. It is mainly used to treat febrile diseases, dysentery caused by heat toxins, carbuncles and boils. When used with dandelion, violet, wild chrysanthemum, etc., it can enhance the heat-clearing and detoxifying effect. When used with coptis, purslane, etc., it has the effect of clearing heat and regulating the intestines, resolving stagnation and promoting blood circulation. It can be used for damp-heat obstruction, damage to the intestinal lining, dysentery with purulent blood, more blood than pus, abdominal pain, tenesmus. When used with coptis, red peony, peony bark, phellodendron, pulsatilla, etc., it has the effect of clearing heat and detoxifying, cooling blood and stopping dysentery. It can be used for epidemic toxins invading the stomach and intestines, stagnation of qi and blood, dysentery with fresh purple purulent blood, high fever, thirst, restlessness, and even delirium.

[0017] Honeysuckle contains pharmacologically active ingredients such as chlorogenic acid and luteolin glycosides, which have strong inhibitory effects on various pathogenic bacteria, including hemolytic streptococci, Staphylococcus aureus, Escherichia coli, Shigella dysenteriae, Vibrio cholerae, Salmonella typhi, and Salmonella paratyphi, as well as viruses causing upper respiratory tract infections. In addition, it also has anti-inflammatory and antipyretic effects, enhances immune function, stops bleeding (coagulation), and resists endotoxins. When used in combination with forsythia, its antibacterial spectrum can be complementary. When used in combination with penicillin, it can enhance the antibacterial effect of penicillin against drug-resistant Staphylococcus aureus.

[0018] Forsythia: It tastes bitter and is slightly cold. It enters the lung, heart, and small intestine meridians. It has the functions of clearing heat and detoxifying, dispersing nodules and reducing swelling. It has an inhibitory effect on Salmonella typhi, Salmonella paratyphi, Escherichia coli, Shigella dysenteriae, Corynebacterium diphtheriae, Vibrio cholerae, Staphylococcus aureus, Streptococcus, etc.

[0019] Scutellaria baicalensis: Bitter in taste and cold in nature, it enters the lung, gallbladder, spleen, stomach, and large intestine meridians. It has the functions of clearing heat and drying dampness, purging fire and detoxifying, stopping bleeding, and calming the fetus. This herb has a wide range of effects and can be used for all damp-heat syndromes. It mainly enters the upper jiao (lung) meridian, primarily clearing lung fire. It is effective for damp-heat diarrhea, lung-heat cough, high fever and thirst, carbuncles and boils. Pharmacological studies have shown that Scutellaria baicalensis contains flavonoids such as baicalin and baicalein, which have inhibitory effects on various pathogenic bacteria and also have anti-inflammatory, antiviral, hepatoprotective, and choleretic effects. When combined with Coptis chinensis and Forsythia suspensa, it can enhance the effects of clearing heat and detoxifying, drying dampness and stopping diarrhea.

[0020] Coptis chinensis: Bitter in taste, cold in nature, and enters the heart, spleen, stomach, liver, gallbladder, and large intestine meridians. It has the functions of clearing heat and drying dampness, purging fire and detoxifying. This herb is particularly effective in clearing heat from the heart and stomach, and also treats damp-heat in the intestines and stomach. It is one of the most potent heat-clearing herbs among traditional Chinese medicines for clearing heat and detoxifying. It is suitable for damp-heat diarrhea, high fever with delirium, restlessness and insomnia, carbuncles, boils, and other symptoms. Pharmacological studies have shown that berberine and other components contained in Coptis chinensis have significant inhibitory effects on various pathogenic bacteria such as Staphylococcus aureus, hemolytic streptococci, and Shigella dysenteriae. It also possesses anti-inflammatory, choleretic, and hypoglycemic pharmacological activities. When used in combination with Scutellaria baicalensis, its heat-clearing and dampness-drying effects are doubled, making it an essential medicine for treating damp-heat diarrhea.

[0021] Dandelion: Bitter and sweet in taste, cold in nature, and enters the liver and stomach meridians. It has the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules, and promoting urination. It is effective in treating boils, carbuncles, mastitis, lung abscesses, intestinal abscesses, damp-heat jaundice, and painful urination due to heat. Dandelion is known as a "key medicine for mastitis" and has a good effect on boils and carbuncles. Pharmacological studies have shown that dandelion decoction has a strong bactericidal effect on Staphylococcus aureus (including drug-resistant strains), hemolytic streptococci, and other pathogenic bacteria. When used together with honeysuckle, violet, and wild chrysanthemum, it can significantly enhance the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules.

[0022] Wild chrysanthemum: Bitter and pungent in taste, slightly cold in nature, and enters the liver and heart meridians. It has the effects of clearing heat and detoxifying, purging fire and calming the liver. It is commonly used to treat boils, carbuncles, red and swollen eyes, sore throat, headache, and dizziness. Wild chrysanthemum is particularly effective at clearing heat and detoxifying, with a strong detoxifying effect. Modern pharmacological research shows that the flavonoids in wild chrysanthemum have antibacterial and antiviral effects, effectively inhibiting the growth and reproduction of pathogens and promoting the phagocytosis of Staphylococcus aureus by white blood cells. When combined with honeysuckle, dandelion, and violet, it works synergistically to clear heat and detoxify, reduce swelling and dissipate nodules.

[0023] Viola yedoensis: Bitter and pungent in taste, cold in nature, and enters the heart and liver meridians. It has the effects of clearing heat and detoxifying, cooling blood and reducing swelling. This herb primarily enters the blood aspect of the heart and liver, effectively cooling the blood and dispersing blood heat stagnation, making it a key medicine for treating boils and carbuncles. It is mainly used to treat boils, carbuncles, erysipelas, and other conditions caused by blood heat stagnation. Pharmacological studies have shown that Viola yedoensis has antimicrobial and anti-inflammatory effects. When used in combination with dandelion, it effectively treats boils and carbuncles; when combined with honeysuckle, its effects of clearing heat and detoxifying, cooling blood and reducing swelling are enhanced. In this formula, dandelion and wild chrysanthemum serve as adjuvant herbs, enhancing its detoxifying, pus-draining, antibacterial, and anti-inflammatory effects.

[0024] Licorice root: Sweet in taste, neutral in nature, and enters the heart, lung, spleen, and stomach meridians. It has the effects of tonifying the spleen and replenishing qi, clearing heat and detoxifying, resolving phlegm and relieving cough, relieving spasms and pain, and harmonizing the properties of other herbs. In traditional Chinese medicine, it is said that "nine out of ten prescriptions contain licorice root," making it one of the most commonly used herbs for harmonizing medicinal properties. Licorice root can both tonify the qi of the spleen and stomach and alleviate the toxicity and harshness of other drugs. Pharmacological studies have shown that licorice root has anti-inflammatory, antiviral, detoxifying, and immunomodulatory effects. In this prescription, licorice root tonifies qi and harmonizes the middle jiao, harmonizes the properties of other herbs, mitigates the harshness of other cold and cooling herbs, and takes into account the delicate physiological characteristics of parrots, serving as the guiding herb. (Explanation of the Chinese herbal formula:)

[0025] Honeysuckle, with its light and fragrant texture, clears heat from the Qi level and removes toxins from the blood level. Forsythia, clearing heat, detoxifying, dispersing nodules, and reducing swelling, serves as the principal herb. Scutellaria and Coptis, bitter and cold in nature, clear heat, dry dampness, and stop diarrhea, enhancing the heat-clearing power of the principal herbs, thus serving as assistant herbs. Dandelion, wild chrysanthemum, and Viola yedoensis, with their medicinal properties, enhance detoxification, pus drainage, antibacterial, and anti-inflammatory effects, serving as adjuvant herbs. Licorice, tonifying Qi and harmonizing the middle Jiao, harmonizes the effects of other cold and cooling herbs, and moderates the harshness of other herbs, while also considering the delicate physiological characteristics of parrots, serving as the guiding herb. Together, they achieve the effect of clearing heat, cooling blood, and stopping dysentery.

[0026] This invention utilizes eight traditional Chinese medicines—honeysuckle, coptis, scutellaria, forsythia, dandelion, wild chrysanthemum, viola, and licorice—in the above-mentioned proportions for the treatment of Staphylococcus psittaci infection. It focuses on targeted treatment of the core pathogenesis of heat-toxin while also considering the physiological characteristics and constitution of psittaci. It leverages the antibacterial effects of each herb while achieving synergistic effects through scientific formulation. The combined action of these herbs comprehensively eliminates the pathological products of Staphylococcus infection, such as redness, swelling, heat, pain, suppuration, and necrosis, systematically resolving the clinical symptoms of Staphylococcus psittaci infection.

[0027] The traditional Chinese medicine composition of the present invention may also contain pharmaceutically acceptable excipients, such as commonly used pharmaceutical carriers, excipients, diluents, etc., and be prepared into various dosage forms such as oral liquids, powders, granules, pills, and capsules according to conventional preparation methods.

[0028] A method for preparing a traditional Chinese medicine composition for treating Staphylococcus psittaci infection includes the following steps:

[0029] (1) Weigh out the following ingredients by weight: honeysuckle, forsythia, scutellaria, coptis, dandelion, wild chrysanthemum, violet, and licorice. Clean them to remove impurities.

[0030] (2) After mixing the above-mentioned medicinal materials, add 6-12 times the weight of the medicinal materials in water and soak for 30-60 minutes;

[0031] (3) Decoction and extraction twice. The first decoction is for 1-2 hours and the second decoction is for 0.5-1 hour. The decoctions are combined, filtered, and the extract and residue are obtained.

[0032] (4) Concentrate the filtrate to a thick paste with a relative density of 1.10-1.25 (measured at 60℃), add an appropriate amount of preservative, sterilize and dispense to obtain oral liquid.

[0033] As a further aspect of the present invention: the decoction extraction in step (3) preferably adopts the following conditions: the first decoction is made with 10 times the amount of water and decocted for 1.5 hours; the second decoction is made with 8 times the amount of water and decocted for 45 minutes.

[0034] As a further embodiment of the present invention, the preparation method further includes drying, pulverizing and granulating the thick paste obtained in step (4) to make granules or powders.

[0035] Compared with the prior art, the beneficial effects of the present invention are:

[0036] 1. This invention, based on traditional Chinese veterinary medicine theory, targets the core pathogenesis of Staphylococcus psittaci infection—"excessive heat and toxins, stagnation of qi and blood, and swelling and pain from sores"—and establishes the treatment principle of "clearing heat and detoxifying, reducing swelling and draining pus, cooling blood and dispersing blood stasis, and supporting the body's resistance to pathogens." The formula uses honeysuckle and forsythia as the principal herbs, which synergistically exert significant antibacterial and anti-inflammatory effects; scutellaria and coptis are the assistant herbs, which are bitter and cold, clearing heat, drying dampness and detoxifying, and enhancing the heat-clearing power of the principal herbs; dandelion, wild chrysanthemum, and violet are the adjuvant herbs, clearing heat and detoxifying, reducing swelling and dispersing nodules; licorice is the guiding herb, tonifying qi and harmonizing the middle jiao, and coordinating the effects of all the herbs. The synergistic combination of these herbs achieves the effects of clearing heat and detoxifying, reducing swelling and dispersing nodules.

[0037] 2. The traditional Chinese medicine preparation of this invention has a significant inhibitory effect on Staphylococcus aureus. In vitro antibacterial tests show that its minimum inhibitory concentration (MIC) against Staphylococcus aureus is 0.5 mg / mL, and its minimum bactericidal concentration (MBC) is 1.0 mg / mL. Clinical treatment trials show that this preparation has a cure rate of 85.0% and a total effective rate of 95.0% for Staphylococcus psittaci infection, which is superior to doxycycline hydrochloride, and has no toxic side effects and high safety. Detailed Implementation

[0038] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0039] Example 1

[0040] Weigh out 15g of honeysuckle, 12g of forsythia, 9g of scutellaria, 6g of coptis, 15g of dandelion, 9g of wild chrysanthemum, 12g of violet, and 6g of licorice. Clean and remove impurities. Mix the above herbs and add 10 times (840mL) of water, then soak for 45 minutes. Extract twice by decoction: first decoction for 1.5 hours, filter the decoction; second decoction with 8 times (672mL) of water for 45 minutes, combine the two decoctions. Concentrate the filtrate to a thick paste with a relative density of 1.15 (60℃), add 0.1% sodium benzoate as a preservative, sterilize, and dispense into 10mL glass bottles, seal and store to obtain the oral liquid product.

[0041] Example 2

[0042] Weigh out 12g of honeysuckle, 10g of forsythia, 8g of scutellaria, 5g of coptis, 12g of dandelion, 8g of wild chrysanthemum, 10g of violet, and 4g of licorice. The preparation method is the same as in Example 1.

[0043] Example 3

[0044] Weigh out 18g of honeysuckle, 14g of forsythia, 10g of scutellaria, 7g of coptis, 16g of dandelion, 10g of wild chrysanthemum, 12g of violet, and 5g of licorice. The preparation method is the same as in Example 1.

[0045] Example 4

[0046] Weigh out 15kg of honeysuckle, 12kg of forsythia, 9kg of scutellaria, 6kg of coptis, 15kg of dandelion, 9kg of wild chrysanthemum, 12kg of violet, and 6kg of licorice. Clean and remove impurities, then add 10 times the amount of water and soak for 45 minutes. Decoction is performed twice (1.5 hours the first time, 45 minutes the second time). The decoctions are combined and concentrated to a thick paste with a relative density of 1.20 (60℃). Add appropriate amounts of dextrin and sucrose powder, mix well, granulate, dry, sizing, and package into 5g granules per bag.

[0047] Example 5

[0048] Weigh out 15g of honeysuckle, 12g of forsythia, 9g of scutellaria, 6g of coptis, 15g of dandelion, 9g of wild chrysanthemum, 12g of violet, and 6g of licorice. Clean and select the herbs, then dry them in an oven at 60℃ until constant weight. Grind each herb into powder and pass it through an 80-mesh sieve. Mix the powders thoroughly according to the weight ratio, divide them into 3g portions, and seal them for storage in aluminum foil bags.

[0049] Comparative Example 1

[0050] A comparative composition was prepared according to the formula of Example 1 in Chinese Patent CN201811627013: 15 parts of Isatis root, 8 parts of Cyperus rhizome, 10 parts of Magnolia bark, 10 parts of Plantago seed, 10 parts of Dandelion, 8 parts of Scutellaria root, 15 parts of Cirsium japonicum, 8 parts of dried tangerine peel, and 10 parts of rhubarb (converted to grams by weight). The preparation method follows the steps in the patent specification.

[0051] Comparative Example 2

[0052] A comparative composition was prepared according to the formulation of the example in Chinese patent CN114681548A: 8 parts of stir-fried Atractylodis Macrocephalae, 8 parts of stir-fried Chinese yam, 5 parts of red halloysite, 5 parts of poppy husk, and 3 parts of plantain seed (converted to grams by weight parts), and the preparation method was referred to the specification of said patent.

[0053] Comparative Example 3 (Composition without Coptidis Rhizoma)

[0054] 15 g of Lonicerae Japonicae Flos, 12 g of Forsythiae Fructus, 9 g of Scutellariae Radix, 15 g of Taraxaci Herba, 9 g of Chrysanthemi Indici Flos, 12 g of Violae Herba, and 6 g of Glycyrrhizae Radix et Rhizoma, and the preparation method was the same as that of Example 1.

[0055] Comparative Example 4 (Composition without Lonicerae Japonicae Flos and Forsythiae Fructus)

[0056] 9 g of Scutellariae Radix, 6 g of Coptidis Rhizoma, 15 g of Taraxaci Herba, 9 g of Chrysanthemi Indici Flos, 12 g of Violae Herba, and 6 g of Glycyrrhizae Radix et Rhizoma, and the preparation method was the same as that of Example 1.

[0057] Test Example 1 (Fractional Inhibitory Concentration Index Test)

[0058] 1.1 Test Method

[0059] The micro-broth checkerboard dilution method was used to determine the antibacterial effect of pairwise combinations of the four main traditional Chinese medicines, namely Lonicerae Japonicae Flos, Forsythiae Fructus, Scutellariae Radix and Coptidis Rhizoma, against Staphylococcus aureus. The fractional inhibitory concentration index (FIC index) was calculated to evaluate the nature of the interaction between each drug pair. An FIC index ≤ 0.5 indicates synergistic interaction, 0.5 < FIC ≤ 1 indicates additive interaction, 1 < FIC ≤ 4 indicates irrelevant interaction, and FIC > 4 indicates antagonistic interaction.

[0060] 1.2 Test Results

[0061] The combined antibacterial effect of each drug pair is shown in Table 3.

[0062] Table 1 Combined antibacterial effect of each drug pair against Staphylococcus aureus

[0063] Honeysuckle + Forsythia 0.375 Synergistic effect Honeysuckle + Scutellaria baicalensis 0.50 Synergistic effect Honeysuckle + Coptis 0.375 Synergistic effect Forsythia + Scutellaria 0.75 Additive effect Forsythia suspensa + Coptis chinensis 0.50 Synergistic effect Scutellaria baicalensis + Coptis chinensis 0.375 Synergistic effect

[0064] The combined antibacterial test results show that the drug pairs of Lonicerae Japonicae Flos-Forsythiae Fructus, Lonicerae Japonicae Flos-Coptidis Rhizoma, Forsythiae Fructus-Coptidis Rhizoma, and Scutellariae Radix-Coptidis Rhizoma all show synergistic interaction; Lonicerae Japonicae Flos-Scutellariae Radix shows synergistic interaction, and Forsythiae Fructus-Scutellariae Radix shows additive interaction. The results indicate that the reasonable compatibility of the main active traditional Chinese medicinal materials in the composition of the present invention can produce a synergistic antibacterial effect, which further confirms the scientificity and advancement of the formulation of the present invention.

[0065] Test Example 2 (In vitro antibacterial test)

[0066] 2.1 Test Materials

[0067] Test strain: Staphylococcus aureus standard strain ATCC25923, purchased from the China Institute of Veterinary Drug Control.

[0068] Test samples: the traditional Chinese medicine composition prepared in Example 1 (sample A), the composition prepared in Comparative Example 1 (sample B), the composition prepared in Comparative Example 2 (sample C), the composition prepared in Comparative Example 3 (sample D), the composition prepared in Comparative Example 4 (sample E), decoction of honeysuckle, decoction of forsythia, decoction of scutellaria, and decoction of coptis.

[0069] Positive control: Berberine hydrochloride (purity ≥98%, Sigma).

[0070] Culture media: MH broth medium and MH agar medium, purchased from Beijing Luqiao Technology Co., Ltd.

[0071] 2.2 Test Methods

[0072] The minimum inhibitory concentration (MIC) of each test sample against Staphylococcus aureus was determined using the broth two-fold microdilution method. Each test sample was serially diluted twofold with MH broth medium and then added to 96-well plates, 100 μL per well. The volume was adjusted to 1 × 10⁻⁶. 6 100 μL of CFU / mL bacterial suspension was added to each well to make a total volume of 200 μL. Positive control wells (containing only bacterial suspension) and negative control wells (containing only culture medium) were included. After incubation at 37°C for 18-24 hours, the lowest drug concentration at which no bacterial growth was observed visually was defined as the MIC. Simultaneously, the minimum bactericidal concentration (MBC) was determined using the plate spread method. Culture media from wells with values ​​above the MIC were spread onto MH agar plates and incubated at 37°C for 24 hours. The corresponding drug concentration with fewer than 5 colonies was defined as the MBC.

[0073] 2.3 Test Results

[0074] The MIC and MBC results for each sample against Staphylococcus aureus are shown in Table 1.

[0075] Table 2. MIC and MBC of different samples against Staphylococcus aureus

[0076] Example 1 0.5 1.0 Example 2 0.5 1.0 Example 3 0.5 1.0 Comparative Example 1 31.25 62.5 Comparative Example 2 62.5 125 Comparative Example 3 1.0 2.0 Comparative Example 4 2.0 4.0 Honeysuckle (single flavor) 4.6875~6.25 9.375~12.5 Forsythia suspensa (single flavor) 31.25 62.5 Scutellaria baicalensis (single herb) 1.25 2.50 Coptis chinensis alone (berberine hydrochloride) 0.128 (calculated as berberine) 0.256 (calculated as berberine)

[0077] As shown in Table 2, the MIC (microinhibitory concentration) of the traditional Chinese medicine compositions in Examples 1-3 of this invention against Staphylococcus aureus is 0.5 mg / mL, and the MBC (microinhibitory concentration) is 1.0 mg / mL, demonstrating significantly better antibacterial effects than the existing patented compositions in Comparative Examples 1 and 2. The MIC of Comparative Example 1 is 31.25 mg / mL, and the MIC of Comparative Example 2 is 62.5 mg / mL. The antibacterial activity of the traditional Chinese medicine compositions of this invention is 62.5 times that of Comparative Example 1 and 125 times that of Comparative Example 2, respectively.

[0078] Compared with the composition lacking a single herb, the antibacterial effect of Example 1 (complete formula) was twice that of Comparative Example 3 (lacking Coptis chinensis) (MIC 0.5 vs 1.0) and four times that of Comparative Example 4 (lacking Lonicera japonica and Forsythia suspensa) (MIC 0.5 vs 2.0), indicating that the Coptis chinensis and Lonicera japonica-Forsythia suspensa herb pair played a key role in the overall formula, and there was a significant synergistic effect among the various Chinese herbal medicines.

[0079] Furthermore, the MBC of the traditional Chinese medicine composition of the present invention against Staphylococcus aureus is 1.0 mg / mL, which is much lower than the MBC value of the prior art compositions (62.5 mg / mL for Comparative Example 1 and 125 mg / mL for Comparative Example 2), indicating that the bactericidal effect of the present invention is more thorough.

[0080] Test Example 3 (Safety Test)

[0081] 3.1 Acute toxicity test

[0082] Forty healthy adult budgies (weighing 35±5g) were randomly divided into four groups of ten each. Example 1 group consisted of three dosage groups: high, medium, and low. The budgies were administered orally once at 10 times (5.0mL / kg), 5 times (2.5mL / kg), and 2.5 times (1.25mL / kg) the clinically recommended dose (0.5mL / kg·d), respectively. The control group received an equal volume of physiological saline. The parrots were observed for seven consecutive days, and their mental state, appetite, activity, defecation, and mortality were recorded.

[0083] The results showed that no deaths occurred in any of the treatment groups within 7 days after administration, and no abnormalities were observed in mental state, appetite, activity, or defecation. In the high-dose group (5.0 mL / kg, equivalent to 10 times the clinical dose), 3 cases of diarrhea and 2 cases of decreased appetite occurred within 24 hours after administration; these recovered spontaneously after 48 hours, with no deaths. No significant abnormalities were observed in the medium- and low-dose groups. This indicates that the compound has good safety within 5 times the clinical dose.

[0084] 3.2 Long-term toxicity test

[0085] Forty healthy adult budgies (35±5g) were randomly divided into two groups of 20 each. In Example 1, the treatment group received the clinically recommended dose (0.5mL / kg·d) orally for 28 consecutive days; the control group received an equal volume of physiological saline. General behavioral indicators were recorded during the observation period. Animals were sacrificed 24 hours after the last administration, and blood samples were collected for complete blood count and liver and kidney function tests. Pathological changes in major organs (heart, liver, spleen, lungs, and kidneys) were also observed through dissection.

[0086] The results showed that the parrots in the treatment group exhibited no abnormalities in mental state, appetite, or activity during the 28-day treatment period. Blood routine tests and liver and kidney function indicators (ALT, AST, BUN, Cr) showed no significant differences compared to the blank control group (P>0.05). Gross examination after dissection revealed no abnormal pathological changes in any of the major organs. This indicates that long-term administration of the herbal composition of this invention has no significant toxic side effects on parrots.

[0087] Trial Case 4 (Clinical Treatment Trial)

[0088] 4.1 Test Subjects

[0089] Sixty parrots diagnosed with Staphylococcus aureus infection by pathogens and admitted to Ruipai Pet Hospital (Baiyun Avenue North Branch) in Baiyun District, Guangzhou City, Guangdong Province between March and May 2024 were randomly divided into three groups of 20 parrots each: Example 1 treatment group, Comparative Example 1 control group, and positive drug control group.

[0090] 4.2 Administration method

[0091] Example 1 group: Take the oral liquid prepared in Example 1 of the present invention, add 0.5 mL per kilogram of body weight, dilute with water and drink freely, twice a day for 7 consecutive days.

[0092] Comparative Example 1: Take the composition of Comparative Example 1, calculate the equivalent dose for each parrot, dilute it with water for free drinking, twice a day for 7 consecutive days.

[0093] Positive drug group: Doxycycline hydrochloride, administered orally at a dose of 10 mg / kg body weight, once daily for 7 consecutive days.

[0094] 4.3 Criteria for Evaluating Therapeutic Effect

[0095] Cure: Clinical symptoms completely disappear, mental state is good, appetite returns to normal, joint swelling subsides, activity is normal, and the pathogen test for Staphylococcus aureus turns negative.

[0096] Effective: Clinical symptoms significantly improved, mental state improved, appetite basically restored, and etiological examination showed Staphylococcus aureus to be negative or the bacterial load was significantly reduced.

[0097] Ineffective: Clinical symptoms show no significant improvement or worsen, and etiological examination remains positive.

[0098] Overall effective rate = (number of cured cases + number of effective cases) / total number of cases × 100%.

[0099] 4.4 Test Results

[0100] The statistical results of the efficacy of each group after 7 days of treatment are shown in Table 2.

[0101] Table 3 Comparison of clinical treatment effects among groups

[0102] Example 1 Group 20 17 19 1 85.0% 95.0% Comparative Example 1 20 8 13 7 40.0% 65.0% Positive drug group 20 15 18 2 75.0% 90.0%

[0103] As shown in Table 3, the total effective rate of the treatment group in Example 1 of this invention was 95.0%, and the cure rate was 85.0%, which was comparable to the efficacy of the positive drug doxycycline hydrochloride group (both had a total effective rate of 95.0%, P>0.05), and significantly better than that of the control group 1 (total effective rate 65.0%, P<0.01). It is noteworthy that no toxic side effects occurred in the Example 1 group during treatment, while two parrots in the positive drug group experienced mild gastrointestinal reactions such as loss of appetite. During a two-week follow-up after drug withdrawal, none of the 17 cured parrots in the Example 1 group relapsed, while one parrot in the positive drug group relapsed. This indicates that the herbal composition of this invention has a stable and lasting therapeutic effect, and superior safety.

[0104] Industrial applicability:

[0105] The traditional Chinese medicine composition of this invention has a simple preparation process, low cost, and is suitable for industrial production. This composition can be used to produce veterinary drug preparations for treating Staphylococcus psittaci infections, offering significant economic and social benefits.

[0106] The above description is merely a preferred embodiment of the present invention and does not limit the scope of the patent. Any equivalent structural or procedural modifications made based on the content of this specification, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of this invention.

[0107] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for treating Staphylococcus psittaci infection, characterized in that: It is made from the following raw materials in parts by weight: honeysuckle 10-20 parts, forsythia 8-16 parts, scutellaria baicalensis 6-12 parts, coptis chinensis 4-8 parts, dandelion 10-18 parts, wild chrysanthemum 6-12 parts, violet 8-14 parts, and licorice 3-8 parts.

2. The traditional Chinese medicine composition for treating Staphylococcus psittaci infection according to claim 1, characterized in that: It is made from the following raw materials in parts by weight: honeysuckle 12-18 parts, forsythia 10-14 parts, scutellaria baicalensis 8-10 parts, coptis chinensis 5-7 parts, dandelion 12-16 parts, wild chrysanthemum 8-10 parts, violet 10-12 parts, and licorice 4-6 parts.

3. The traditional Chinese medicine composition for treating Staphylococcus psittaci infection according to claim 2 is characterized in that: It is made from the following raw materials in parts by weight: honeysuckle 15 parts, forsythia 12 parts, scutellaria baicalensis 9 parts, coptis chinensis 6 parts, dandelion 15 parts, wild chrysanthemum 9 parts, viola yedoensis 12 parts, and licorice 6 parts.

4. A method for preparing a traditional Chinese medicine composition for treating Staphylococcus psittaci infection as described in any one of claims 1-3, characterized in that: Includes the following steps: Step (1): Weigh each raw material according to the weight parts and clean it to remove impurities; Step (2): Mix the above-mentioned medicinal materials and add water at a ratio of 6-12 times the total weight of the medicinal materials, and soak for 30-60 minutes; Step (3): Decoction and extraction twice. The first decoction is for 1-2 hours and the second decoction is for 0.5-1 hour. Combine the decoctions, filter, and obtain the extract and dregs. Step (4): Concentrate the filtrate to a thick paste with a relative density of 1.10-1.25 (measured at 60℃).

5. The method for preparing a traditional Chinese medicine composition for treating Staphylococcus psittaci infection according to claim 4, characterized in that: In step (3), add 10 times the amount of water for the first decoction and decoct for 1.5 hours; add 8 times the amount of water for the second decoction and decoct for 45 minutes.

6. The method for preparing a traditional Chinese medicine composition for treating Staphylococcus psittaci infection according to claim 4, characterized in that: Step (4) further includes drying, pulverizing, and granulating the viscous paste to produce powder or granules.

7. The method for preparing a traditional Chinese medicine composition for treating Staphylococcus psittaci infection according to claim 4, characterized in that: Add a pharmaceutically acceptable preservative to the thick paste obtained in step (4), sterilize it, and then package it into an oral liquid.

8. The use of the traditional Chinese medicine composition according to any one of claims 1-3 in the preparation of a medicament for treating Staphylococcus psittaci infection.