A compound for preventing and treating poultry bacterial diseases and its application

Abstract

The present invention discloses a compound for preventing and treating bacterial diseases in poultry and its use, relating to the technical field of veterinary medicine. The compound comprises the following components by weight: 10-20 parts of an astragalus root extract, 4-10 parts of a chestnut shell extract, and 1-5 parts of a peach leaf extract. The raw medicinal materials selected in the present invention, when used together, have the functions of clearing heat and detoxifying, replenishing qi and raising yang, providing antibacterial and anti-inflammatory effects, and enhancing immunity. It has a good preventive and therapeutic effect against common bacterial diseases in poultry, is not prone to drug resistance, and does not produce drug residues that are harmful to human health. Furthermore, the raw materials are readily available and low-cost, making it suitable for large-scale breeding.

Description

Technical Field

[0001] The present invention relates to the technical field of veterinary medicines, and in particular to a compound for preventing and treating poultry bacterial diseases and application thereof. Background Art

[0002] With the development of my country's agriculture and rural economy, large-scale poultry farming has become an important way for farmers to increase their income and become wealthy. The gradual evolution of poultry farming from free-range to large-scale operations has brought significant economic benefits to farmers and driven the rapid development of the poultry industry. Disease prevention and control are crucial in poultry farming. Bacterial diseases, such as Escherichia coli, Salmonellosis, and Staphylococcus aureus, are the most common diseases affecting poultry. Currently, the majority of drugs used to prevent and treat bacterial diseases in poultry are antibiotics and chemically synthesized antimicrobials, such as enrofloxacin, streptomycin, gentamicin, amoxicillin, and sulfonamides. With the widespread clinical use of antimicrobial drugs, drug resistance is becoming increasingly serious, with the emergence of multidrug-resistant strains and super-resistant bacteria, posing significant risks to human and animal health. Bacterial resistance has become a major concern for public health and agricultural product safety.

[0003] Faced with the increasingly severe threat of bacterial resistance and the increasingly stringent use of antibiotics, finding effective traditional Chinese medicines for bacterial control is a viable approach. my country has a long history of using herbal medicine to prevent and treat poultry diseases. Records of using herbal medicine to treat poultry diseases date back to ancient times, and numerous traditional remedies have been accumulated, with remarkable effectiveness. Herbal medicines are rich in proteins, amino acids, vitamins, trace elements, polysaccharides, organic acids, and a variety of naturally active ingredients. Adding them to feed can supplement the nutritional needs of livestock and poultry. Herbal medicines offer the potential for combination and bidirectional regulation, resulting in minimal harmful residues in animals, minimal toxic side effects, and a low risk of pathogenic microorganisms developing resistance. Astragalus membranaceus (Huangqi) has a long history of use in my country and is one of the earliest traditional Chinese medicines for tonifying the middle qi and replenishing qi. A perennial plant of the grass family, it is widely distributed and used worldwide. Due to its anti-inflammatory, antiviral, antioxidant, and immune-enhancing properties, it has long been used in my country and Southeast Asia to treat human and animal diseases. Chestnut is a plant of the Fagaceae family, a tree-like nut-like economic plant widely distributed in Asia, America, and Europe. my country ranks first in the world in both chestnut cultivation area and production. Chestnut processing produces a large amount of chestnut shells, which are primarily discarded through burning or disposal. Chestnut shells have a subtle odor and a slightly bitter and astringent taste. They have the effects of relieving adverse reactions, promoting fluid production, resolving phlegm and relieving cough, clearing heat and dispersing stagnation, and stopping bleeding. They are commonly used to treat nausea, vomiting, thirst, cough with sputum, whooping cough, mumps, scrofula, epistaxis, and bloody stools. Peach leaves contain glycosides, hydroxyl groups, and various organic acids, which have the effects of clearing heat and detoxifying, killing insects, and relieving itching. They are used to treat diseases such as malaria, carbuncles, hemorrhoids, eczema, and vaginal trichomoniasis. Currently, there are no reports of the combined use of astragalus, chestnut shells, and peach leaves for the prevention and treatment of bacterial diseases in poultry. Summary of the Invention

[0004] In view of the above-mentioned prior art, the object of the present invention is to provide a compound for preventing and treating bacterial diseases in poultry and its application.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] The first aspect of the present invention provides a compound for preventing and treating poultry bacterial diseases, comprising the following components in parts by weight: 10-20 parts of astragalus extract, 4-10 parts of chestnut shell extract and 1-5 parts of peach leaf extract.

[0007] Preferably, the complex comprises the following components in parts by mass: 15 parts of astragalus extract, 7 parts of chestnut shell extract and 3 parts of peach leaf extract.

[0008] Preferably, the astragalus extract is obtained by the following method:

[0009] (1) Drying and crushing the root of Astragalus membranaceus and sieving it to obtain Astragalus membranaceus powder, adding water to the Astragalus membranaceus powder and mixing it, ultrasonically extracting it 2-4 times, and filtering it to obtain an aqueous extract;

[0010] (2) Anhydrous ethanol was added to the aqueous extract to adjust the alcohol concentration to 80%, and the mixture was allowed to stand. The supernatant was filtered, concentrated, and dried to obtain the astragalus extract.

[0011] Preferably, in step (2), the material-liquid ratio of astragalus powder to water is 1 g: (15-25) ml, the ultrasonic extraction time is 20-30 min, and the ultrasonic power is 150 W-200 W.

[0012] Preferably, the chestnut shell extract is obtained by the following method:

[0013] (1) drying and grinding chestnut shells and then sieving to obtain chestnut shell powder;

[0014] (2) adding water to the chestnut shell powder and boiling to obtain a mixture, adding cellulase and xylanase after the mixture is cooled, and performing microwave-assisted enzymolysis for 4-7 minutes, and then continuing the enzymolysis. After the enzymolysis is completed, the enzyme is inactivated, and the mixture is cooled to obtain an enzymolysis solution;

[0015] (3) The enzymatic hydrolyzate is centrifuged, and the supernatant is concentrated and dried to obtain a chestnut shell extract.

[0016] Preferably, in step (2), the mass ratio of chestnut shell powder to water is 1:15-20, the amount of cellulase added is 1%-2% of the mass of the mixed solution, and the amount of xylanase added is 1%-2% of the mass of the mixed solution.

[0017] Preferably, in step (2), the conditions for microwave-assisted enzymatic hydrolysis are: extraction at a microwave temperature of 50-60°C and a microwave power of 400-500W for 4-6 times, each time for 15-20 seconds, with an interval of 1 minute between each extraction; and continued enzymatic hydrolysis at a temperature of 45-55°C for 2-3 hours.

[0018] Preferably, the peach leaf extract is obtained by the following method:

[0019] Peach leaf powder and distilled water are mixed in a mass ratio of 1:10-15, heated at 80-100° C. for 10-12 hours, the upper distilled oil is collected, and the mixture is dehydrated with anhydrous sodium sulfate to obtain a peach leaf extract.

[0020] The second aspect of the present invention provides the use of the above-mentioned complex in the preparation of a product for preventing and treating bacterial diseases in poultry.

[0021] Preferably, the bacterial disease is poultry colibacillosis, poultry salmonellosis, poultry staphylococcal disease, poultry dysentery, poultry urinary tract infection or poultry Clostridium perfringens disease.

[0022] Beneficial effects of the present invention:

[0023] The pharmacology of each component in the Chinese medicine composition of the present invention is as follows:

[0024] Astragalus can replenish qi and raise yang, consolidate the exterior and stop sweating, promote diuresis and reduce swelling, promote fluid production and nourish blood, relieve stagnation and numbness, remove toxins and discharge pus, heal sores and promote tissue regeneration. It can also improve immunity, resist fatigue, and lower blood sugar. Chestnut shell has a faint smell and a slightly bitter and astringent taste. It has the effects of lowering adverse reactions, promoting fluid production, resolving phlegm and relieving cough, clearing away heat and dispersing stagnation, and stopping bleeding. It is often used to treat nausea, vomiting, diabetes, cough with sputum, whooping cough, mumps, scrofula, epistaxis, and bloody stools. Peach leaves can clear away heat and detoxify, kill insects and relieve itching, and have antibacterial and anti-inflammatory effects. It can be used for malaria, carbuncles, hemorrhoids, eczema, and vaginal trichomoniasis.

[0025] The raw medicinal materials selected by the present invention have the functions of clearing away heat and detoxifying, replenishing qi and raising yang, antibacterial and anti-inflammatory, and enhancing immunity when used together. They have a good preventive and therapeutic effect on common bacterial diseases in poultry, are not prone to drug resistance, and will not produce drug residues that are harmful to human health. In addition, the raw materials are easily available and low in cost, which is conducive to large-scale breeding and use. DETAILED DESCRIPTION

[0026] It should be noted that the following detailed descriptions are illustrative and intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present application belongs.

[0027] In order to enable those skilled in the art to more clearly understand the technical solution of the present application, the technical solution of the present application will be described in detail below with reference to specific embodiments.

[0028] The test materials used in the examples of the present invention are all conventional test materials in the art and can be purchased through commercial channels.

[0029] The strains used in the experiment: Escherichia coli was purchased from the China Center for Type Culture Collection with the accession number CCTCC AB 2012886; Salmonella sp. was purchased from the China Center for Type Culture Collection with the accession number CCTCC CB 20082511; Staphylococcus aureus was purchased from the China Center for Type Culture Collection with the accession number CCTCC CB 20081848; Shigella coli was purchased from the China Center for Type Culture Collection with the accession number CCTCC AB 91116; Proteus mirabilis was purchased from the China Center for Type Culture Collection with the accession number CCTCC GB 2008263; Clostridium perfringens was purchased from the China Center for Type Culture Collection with the accession number CCTCC GB 2008263; perfringens) was purchased from China Industrial Microorganism Culture Collection Center with the collection number CICC25011.

[0030] The cellulase used in the present invention was purchased from Shandong Pingju Biotechnology Co., Ltd., item number: 291, purity: ≥99.9%.

[0031] The xylanase used in the present invention was purchased from Shandong Pingju Biotechnology Co., Ltd., item number: 169, purity: ≥99.9%.

[0032] Example 1: Preparation of a compound for preventing and treating bacterial diseases in poultry

[0033] 1. Raw material composition (parts by weight): 10 parts of Astragalus extract, 4 parts of Chestnut shell extract, and 1 part of Peach Leaf extract;

[0034] 2. Preparation steps:

[0035] (1) Preparation of Astragalus Extract:

[0036] The root of Astragalus membranaceus is dried, crushed, and sieved to obtain Astragalus membranaceus powder; water is added to the Astragalus membranaceus powder and mixed evenly, with the solid-liquid ratio of Astragalus membranaceus powder to water being 1g:20ml; ultrasonic extraction is performed four times, with the ultrasonic extraction time being 25 minutes and the ultrasonic power being 200W; and filtration is performed to obtain an aqueous extract; anhydrous ethanol is added to the aqueous extract to adjust the alcohol concentration to 80%, the extract is allowed to stand, and the supernatant is filtered, concentrated, and dried to obtain an Astragalus membranaceus extract.

[0037] (2) Preparation of chestnut shell extract:

[0038] The chestnut shells were dried, crushed, and sieved to obtain chestnut shell powder; water was added to the chestnut shell powder and boiled to obtain a mixture, wherein the mass ratio of chestnut shell powder to water was 1:18; cellulase and xylanase were added after the mixture was cooled, wherein the amount of cellulase added was 1.5% of the mass of the mixture, and the amount of xylanase added was 1.5% of the mass of the mixture; microwave-assisted enzymatic hydrolysis was performed for 6.5 minutes, and the conditions for microwave-assisted enzymatic hydrolysis were as follows: 6 extractions for 15 seconds each time at a microwave temperature of 55° C. and a microwave power of 500 W, with an interval of 1 minute between each extractions; and then enzymatic hydrolysis was continued at a temperature of 50° C. for 3 hours. After the enzymatic hydrolysis was completed, the enzyme was inactivated and the mixture was cooled to obtain an enzymatic hydrolyzate; the enzymatic hydrolyzate was centrifuged, and the supernatant was concentrated and dried to obtain a chestnut shell extract;

[0039] (3) Preparation of peach leaf extract:

[0040] Peach leaf powder and distilled water were mixed in a mass ratio of 1:12, heated at 85°C for 10 h, the upper distilled oil was collected, and the extract was obtained after dehydration with anhydrous sodium sulfate;

[0041] (4) Astragalus extract, chestnut shell extract, and peach leaf extract are weighed and mixed uniformly according to mass to obtain a complex for preventing and treating poultry bacterial diseases.

[0042] Example 2: Preparation of a compound for preventing and treating bacterial diseases in poultry

[0043] Raw material composition (parts by weight): 15 parts of astragalus extract, 7 parts of chestnut shell extract, 3 parts of peach leaf extract;

[0044] The preparation method is the same as that of Example 1 to prepare a compound for preventing and treating bacterial diseases in poultry.

[0045] Example 3: Preparation of a compound for preventing and treating bacterial diseases in poultry

[0046] Raw material composition (parts by weight): 20 parts of astragalus extract, 10 parts of chestnut shell extract, and 5 parts of peach leaf extract;

[0047] The preparation method is the same as that of Example 1 to prepare a compound for preventing and treating bacterial diseases in poultry.

[0048] Comparative Example 1

[0049] The astragalus extract prepared by the method of Example 1 is used alone as a drug for preventing and treating bacterial diseases in poultry.

[0050] Comparative Example 2

[0051] The chestnut shell extract prepared by the method of Example 1 is used alone as a drug for preventing and treating bacterial diseases in poultry.

[0052] Comparative Example 3

[0053] The peach leaf extract prepared by the method of Example 1 is used alone as a drug for preventing and treating bacterial diseases in poultry.

[0054] Test Example 1: In vitro antibacterial test

[0055] Referring to the step 1.2.2 of the literature "Study on the antibacterial activity of Chuanxiong, dried ginger and Andrographis paniculata extracts and their combination" (Zhao Qing et al. Plant Protection, 2022), experiments were conducted on the complexes for preventing and treating poultry bacterial diseases prepared in Examples 1-3 and the drugs for preventing and treating poultry bacterial diseases of Comparative Examples 1-3.

[0056] The test bacteria species were Escherichia coli, Salmonella sp., Staphylococcus aureus, Shigella coli, Proteus mirabilis, and Clostridium perfringens.

[0057] Frozen bacterial strains were revived at room temperature, and a small number of colonies were picked with an inoculating needle and placed in the center of a PDA plate. The plate was sealed and inverted in an incubator at 25°C and 73% humidity without illumination. The composites for preventing and treating poultry bacterial diseases prepared in Examples 1-3 and the Chinese herbal extracts of Comparative Examples 1-3 were first dissolved in dimethyl sulfoxide (DMSO) and then diluted with 1‰ Tween water to a 500 mg / L solution. 1 mL of each solution was pipetted and added to 9 mL of PDA medium to create a drug-containing plate with a final concentration of 50 mg / L. After activation, Escherichia coli, Salmonella, Staphylococcus aureus, Shigella, Proteus mirabilis, and Clostridium perfringens were punched with a 4 mm diameter bacterial cake and placed on the drug-containing plate, three cakes per plate arranged in an equilateral triangle. Each treatment was repeated three times, with no drug added as a blank control. After culturing each treatment in a (24±1)℃ incubator for 72h, the colony expansion diameter of each treatment was measured and the inhibition rate was calculated.

[0058] The calculation formula of the antibacterial rate is as follows:

[0059] R=(D0-D) / (D0)×100%

[0060] Where: R is the inhibition rate, D0 is the colony diameter of the control group (mm), and D is the colony diameter of the treatment group (mm).

[0061] 2. Test results:

[0062] Table 1 Antibacterial rates of the complexes of Examples 1-3 and the extracts of Comparative Examples 1-3

[0063]

[0064] As can be seen from Table 1, the present invention combines astragalus extract, chestnut shell extract, and peach leaf extract to achieve an inhibition rate of over 85% against Escherichia coli, Salmonella, Staphylococcus aureus, Shigella, Proteus mirabilis, and Clostridium perfringens. This combination can prevent and treat poultry colibacillosis, poultry salmonellosis, poultry staphylococcal disease, poultry dysentery (caused by Shigella), poultry urinary tract infection (caused by Proteus mirabilis), and poultry Clostridium perfringens. The present invention combines astragalus extract, chestnut shell extract, and peach leaf extract to achieve a synergistic bactericidal effect.

[0065] Test Example 2: Clinical Trial

[0066] Four hundred healthy, 30-day-old reed chickens were selected for the study. Each chicken received an intramuscular injection of 0.1 ml of Escherichia coli and 0.1 ml of Salmonella. Clinical symptoms included lethargy, loss of appetite, ruffled feathers, and recumbency; severe diarrhea with grayish-white, watery feces; and fecal contamination around the anus. Two days later, 250 chickens displaying similar symptoms were randomly divided into five groups of 50 birds each.

[0067] Group 1 was treated with the poultry bacterial disease prevention and treatment complex prepared in Example 1 of the present application. Groups 2-4 were treated with the poultry bacterial disease prevention and treatment drugs of Comparative Examples 1-3. The fifth group served as the control group. Treatment methods included administering 1g of the drug per liter of water, administered as a single dose, for 5 consecutive days. The control group received no medication. Following treatment, the chickens were observed for 7 consecutive days. Clinical changes and mortality in each group were recorded, and the overall efficacy and cure rate were calculated. The results are shown in Table 2.

[0068] Clinical efficacy judgment criteria:

[0069] Effective: After the trial, the chickens' disease symptoms, spirits and appetite improved significantly;

[0070] Cure: After the experiment, the symptoms of the sick chickens disappeared, their spirits improved, their appetites increased, and they completely recovered to normal.

[0071] Ineffective: Symptoms appeared during the trial and did not disappear after the trial ended, or corresponding symptoms appeared and died during the experiment.

[0072] Cure rate = number of cured cases / 50×100%

[0073] Total effective rate = (number of cured cases + number of effective cases) / 50 × 100%

[0074] Table 2 The therapeutic effects of different drugs on chicken Escherichia coli and Salmonella infection

[0075]

[0076] As can be seen from Table 2, the effectiveness of the compound for preventing and treating poultry bacterial diseases prepared by the present invention in preventing and treating chicken Escherichia coli and Salmonella infections is 98%, which is 92% higher than that of the blank control group, and has excellent prevention and treatment effects on chicken Escherichia coli and Salmonellosis.

[0077] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A compound for preventing and treating bacterial diseases in poultry, characterized in that: The invention is composed of the following components in parts by mass: 10-20 parts of astragalus extract, 4-10 parts of chestnut shell extract and 1-5 parts of peach leaf extract; The astragalus extract is obtained by the following method: (1) Dry and crush the root of Astragalus membranaceus and sieve it to obtain Astragalus membranaceus powder. Add water to the Astragalus membranaceus powder and mix well. Ultrasonic extraction is performed 2-4 times. Filter and obtain a water extract. The material-liquid ratio of Astragalus membranaceus powder to water is 1g: (15-25)ml. The ultrasonic extraction time is 20-30min and the ultrasonic power is 150W-200W. (2) Adding anhydrous ethanol to the aqueous extract to adjust the alcohol concentration to 80%, allowing the mixture to stand, filtering the supernatant, concentrating, and drying to obtain the astragalus extract; The chestnut shell extract is obtained by the following method: (1) Drying and grinding chestnut shells and then sieving to obtain chestnut shell powder; (2) adding water to the chestnut shell powder and boiling the mixture to obtain a mixture, adding cellulase and xylanase after the mixture is cooled, and performing microwave-assisted enzymolysis for 4-7 minutes, and then continuing the enzymolysis. After the enzymolysis is completed, the enzyme is inactivated, and the mixture is cooled to obtain an enzymolysis solution; (3) centrifuging the enzymatic hydrolyzate, concentrating the supernatant, and drying the supernatant to obtain a chestnut shell extract; The peach leaf extract is obtained by the following method: Peach leaf powder and distilled water are mixed in a mass ratio of 1:10-15, heated at 80-100° C. for 10-12 hours, the upper distilled oil is collected, and the mixture is dehydrated with anhydrous sodium sulfate to obtain a peach leaf extract.

2. The composite according to claim 1, characterized in that The complex is composed of the following components in parts by mass: 15 parts of astragalus root extract, 7 parts of chestnut shell extract and 3 parts of peach leaf extract.

3. The composite according to claim 1, characterized in that The mass ratio of chestnut shell powder to water is 1:15-20, the added amount of cellulase is 1%-2% of the mass of the mixed solution, and the added amount of xylanase is 1%-2% of the mass of the mixed solution.

4. The composite according to claim 1, characterized in that The conditions for microwave-assisted enzymatic hydrolysis are: microwave temperature of 50-60°C, microwave power of 400-500W, extraction 4-6 times, each time for 15-20s, each time with an interval of 1min; continued enzymatic hydrolysis temperature of 45-55°C, enzymatic hydrolysis time of 2-3h.

5. Use of the complex according to claim 1 or 2 in the preparation of products for preventing and treating bacterial diseases in poultry.

6. The use according to claim 5, characterized in that The bacterial disease is poultry colibacillosis, poultry salmonellosis, poultry staphylococcal disease, poultry dysentery, poultry urinary tract infection or poultry Clostridium perfringens disease.

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