Application of Chinese mahonia stem in prevention and treatment of infectious rhinitis of chicken

Abstract

According to the application of the Chinese mahonia stems to prevention and treatment of the infectious rhinitis of the chickens, active ingredients can be composed of single Chinese mahonia stems, the components are simple and easy to prepare, and the Chinese mahonia stems can replace antibiotics to be used in the egg laying period of the chickens and are free of drug residue requirements, high in cure rate, safe, free of toxic and side effects and convenient to use.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary medicines, and particularly relates to a traditional Chinese medicine composition with Chinese mahonia as a core formula for preventing and treating infectious rhinitis in chickens. Background Art

[0002] Infectious rhinitis (ICR), a common acute respiratory infection in poultry farming, is caused by Haemophilus paragallinarum and results in significant economic losses for the poultry industry. Affected birds experience depression, reduced food and water intake, respiratory inflammation, and facial swelling. Egg production in laying hens decreases significantly, and growth in growing chickens is stunted.

[0003] Although sulfonamides are currently effective against Haemophilus paragallinarum, they tend to leave residues in chickens, affecting feed intake and egg production, and are therefore prohibited during the laying period. Traditional Chinese medicine preparations, due to their natural properties, versatility, and low residue requirements, have become a hot research alternative to chemical drugs. However, existing traditional Chinese medicine preparations for the prevention and treatment of infectious rhinitis in chickens have numerous shortcomings. Some traditional Chinese medicine formulas contain a wide variety of medicinal ingredients, making it difficult for small birds to consume sufficient doses to achieve their efficacy. Some traditional Chinese medicine formulas focus solely on the infectious rhinitis itself, lacking effective solutions for complications such as poor growth in growing chickens and decreased egg production in laying hens. Many traditional Chinese medicine formulas contain numerous ingredients, some containing more than a dozen herbs, resulting in complex compositions, high costs, and poor efficacy. These issues urgently need to be addressed to meet the livestock industry's demand for safe and effective treatments for infectious rhinitis in chickens. In traditional Chinese medicine, Chinese magnolia vine is mainly used to treat human diseases. Its functions are to clear away heat and dampness, purge fire and detoxify. It is commonly used to treat damp-heat diarrhea, jaundice and other diseases. Summary of the Invention

[0004] The present invention aims to provide a traditional Chinese medicine composition for preventing and treating infectious rhinitis in chickens, using Chinese mahonia as the sole active ingredient. This composition can replace antibiotics during the laying period of chickens and has the advantages of being free of drug residues, simple to prepare, having a high cure rate, being safe and free of toxic side effects, and being easy to use. Furthermore, it can effectively alleviate problems such as poor growth in growing chickens and decreased egg production in laying hens caused by infectious rhinitis. The specific technical solution is as follows: 1. Formula of Chinese medicine composition: The Chinese medicine composition of the present invention is composed of Mahonia lactiflora or Mahonia lactiflora and Nandina domestica.

[0005] 2. Dosage form: The dosage form is mixture, powder, granules or decoction.

[0006] 3. Preparation method: The present invention further provides a method for preparing the Chinese medicine composition, which comprises the following steps: first, crushing and mixing the medicinal materials to form a powder; and second, extracting the medicinal materials with water to prepare the powder.

[0007] 4. Pharmaceutical use: Use of Chinese magnolia vine in the preparation of drugs for preventing and treating infectious rhinitis in chickens.

[0008] Compared with the prior art, the present invention has the following beneficial technical effects: The present invention selects medicinal materials with antibacterial effects for screening and research, and finds that Mahonia laurel or a combination of Mahonia laurel and Nandina domestica has a significant effect in treating infectious rhinitis in chickens. It can not only effectively inhibit Haemophilus paragallinarum, but also alleviate clinical symptoms, enhance the resistance of chickens, and improve the growth condition of chickens and the egg-laying performance of laying hens. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 0 points for clinical symptoms of chicken infectious coryza; Figure 2 1 point for clinical symptoms of infectious coryza in chickens; Figure 3 2 points for clinical symptoms of chicken infectious coryza; Figure 4 3 points for clinical symptoms of chicken infectious coryza; Figure 5 The chicken farm conditions for infectious coryza in chickens. DETAILED DESCRIPTION

[0010] The specific implementation of the present invention is further explained below with reference to specific examples.

[0011] Example Example 1: Take 1000g of Chinese magnolia vine, crush it, and pass it through a 100-mesh sieve to obtain the product.

[0012] Example 2: Take 1000g of Chinese magnolia vine, add 10 times the amount of water, decoct twice, each time for 1.5 hours, combine the filtrates, and concentrate to contain the crude drug amount of 1g / mL.

[0013] Example 3: Take 500g of Chinese magnolia vine and 500g of Nandina domestica, add 10 times the amount of water, decoct twice, each time for 1.5 hours, combine the filtrates, and concentrate to contain the crude drug amount of 1g / mL.

[0014] Comparative Example 1: Take 1000g of Nandina domestica, add 10 times the amount of water, decoct twice, each time for 1.5 hours, combine the filtrate, and concentrate to contain the crude drug amount of 1g / mL.

[0015] Comparative Example 2: 1000g of Achyranthes bidentata was added with 10 times the amount of water and decocted twice, each time for 1.5 hours. The filtrates were combined and concentrated to contain 1g / mL of crude drug.

[0016] Comparative Example 3: Add 10 times the amount of water to 1000g of Tripterygium wilfordii, decoct twice, each time for 1.5 hours, combine the filtrates, and concentrate to contain 1g / mL of crude drug.

[0017] Comparative Example 4: 1000g of Acanthopanax acanthopanax bark was added with 10 times the amount of water, and the mixture was decocted twice, each time for 1.5 hours. The filtrates were combined and concentrated to contain 1g / mL of crude drug.

[0018] Experimental Example 1 The in vitro anti-Haemophilus influenzae effects of Example 2, Example 3, Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4 were observed.

[0019] In this test, drugs of different concentrations were mixed and dissolved in a liquid culture medium, and bacteria were inoculated. The minimum inhibitory concentration (MIC) of the embodiment was determined based on the growth of the bacteria.

[0020] Experimental materials: Example 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4; Haemophilus paragallinarum type B strain; TSB culture medium; diluent; 37° C. incubator.

[0021] Experimental methods: Prepare a Haemophilus paragallinarum culture medium: Prepare TSB medium by weighing 30g of Tryptic Soy Broth (TSB) and 10g of yeast extract powder. Add 945mL of purified water and shake thoroughly to dissolve. Autoclave at 121°C for 15 minutes. After cooling, add 50mL of inactivated newborn calf serum and 5mL of filter-sterilized 1% NAD solution and shake thoroughly to dissolve. Pick a single Haemophilus paragallinarum colony, add it to the culture medium, and incubate at 37°C for 24 hours.

[0022] Preparation of culture medium containing the example: dilute the drug with physiological saline in multiple ratios to prepare test solutions of different concentrations. Take 2.5 mL of the test solution of each dilution and add it to a test tube containing 2.5 mL of double-concentration TSB culture medium to dilute it to 1:256.

[0023] 20 μl of the bacterial suspension was inoculated into a test tube containing the culture medium of the example as a test group sample.

[0024] The same method was used to inoculate a test tube without the example to serve as a positive control sample.

[0025] Take 2 test tubes containing culture medium and treat them with normal saline instead of drug as negative control group samples.

[0026] The samples of the example group, the positive control group and the negative control group were placed in a 37° C. incubator and cultured for 24 hours. The results were observed. If the positive control group showed obvious turbidity, the culture was stopped and the results were observed.

[0027] Judging Method: When the positive control tube had bacterial growth (turbidity) and the negative control tube had no bacterial growth (transparent), the lowest concentration of the example group with no bacterial growth was the MIC of the sample to the tested bacteria.

[0028] Experimental results: Table 1. Minimum inhibitory concentration (MIC) of different drugs against Haemophilus paragallinarum

[0029] The results showed that the negative control and the positive control were established, and Examples 2 and 3, Comparative Examples 1, 2, 3 and 4 all had in vitro antibacterial effects against Haemophilus paragallinarum, and Nandina domestica or Mahonia macrophylla + Nandina domestica had the best in vitro antibacterial effects.

[0030] Experimental Example 2 Clinical trials on the prevention and treatment of infectious coryza in chickens Cleaning and disinfection of experimental animal rooms: Use formaldehyde fumigation for disinfection, 250g potassium permanganate and 500mL formaldehyde, mixed fumigation for 24 hours; ventilate for 24 hours after disinfection.

[0031] SPF chickens were weighed and grouped: 260 SPF chickens over 30 days old were collected, and those with large weight differences and weak chicks were eliminated. The remaining 240 SPF chickens were divided into 8 groups, with 30 chickens in each group. The weights of the groups were weighed and grouped, and the weight differences between the groups were adjusted to less than 3%, and the data were recorded.

[0032] Preparation of bacterial culture medium TSA plates: Weigh 40 g of Tryptic Soy Agar (TSA) medium, add 845 ml of purified water, shake thoroughly, and heat until fully dissolved. Autoclave at 121°C for 15 min. Cool to approximately 60°C, add 50 mL of healthy, sterile newborn calf serum, 100 mL of yeast extract, and 5 mL of filter-sterilized 1% NAD solution. Shake thoroughly, and pour into a sterile dish to make a plate.

[0033] TSB medium: Weigh 30 g of Tryptic Soy Broth (TSB) medium and 10 g of yeast extract powder, add 945 mL of purified water, shake well to dissolve, and sterilize by autoclaving at 121°C for 15 min. After cooling, add 50 mL of inactivated newborn calf serum and 5 mL of filter-sterilized 1% NAD solution, and shake well to obtain TSB medium.

[0034] Standards for the preparation and control of potent toxins Take freeze-dried Avibacterium paragallinarum type B basic seeds, streak inoculate TSA plates, and culture in a constant temperature incubator at 37°C for 16 to 24 hours. Pick several typical colonies, inoculate them into 100 to 200 mL of TSB medium, and culture at 37°C for 12 to 24 hours. Harvest; dilute 50 times with sterile saline for virus attack and set aside.

[0035] Storage and usage period: Store below -70℃ and the usage period shall not exceed 7 days.

[0036] Model establishment and grouping: Example 1; Comparative Examples 1-4; Model control group; Blank control group. Except for the blank control group, the challenge dose of each group was 1×10 7 cfu / 0.2mL / mouse, the challenge site was intraorbital sinus injection, and the blank control group was injected with normal saline.

[0037] Disease determination criteria: If any of the following conditions are met, the disease is considered to be present: ① discharge of clear to serous viscous secretions from the nostrils, tearing; ② swelling of the infraorbital sinus and surrounding areas on one or both sides of the face. If there are no symptoms, the score is 0. If there are symptoms, the severity of the clinical symptoms of the diseased chickens is observed by comparing them with the blank group. Swelling of the eyes and mucous nasal discharge are scored from mild to severe as 1, 2, or 3 (see Figure 1-4 ).

[0038] After the challenge, the corresponding gavage medication was started for 7 days to ensure that the amount of drug solution administered to each group was consistent.

[0039] After the drug administration, each group was given sufficient food and water.

[0040] Test results: Table 2. Effects of each group on preventing and treating infectious coryza in chickens

[0041] Note: The same letters indicate no significant difference (P≥0.05), and different letters indicate significant difference (P<0.05). The results show that after 7 days of drug use, Examples 2 and 3 have the best effect on reducing swelling caused by Haemophilus paragallinarum in chickens, especially Example 3, which has significant differences compared with the model group. Compared with the solution of the present invention, Comparative Examples 1-4 do not show better effects. It can be seen that medicinal materials with better or worse in vitro antibacterial effects than Example 2 may not necessarily have the effect of effectively treating infectious rhinitis in chickens when taken orally.

[0042] Experimental Example 3 The effect of preventing and treating infectious coryza in chickens of Example 1 was verified by the following cases.

[0043] In order to verify the regional clinical therapeutic effect of the present invention on infectious coryza in chickens, the effect verification experiment of Example 1 was carried out in a farm infected with infectious coryza in chickens; because laying hens cannot use sensitive antibiotics, the experiment was divided into Example 1 and a negative control group, and the effects were compared after 7 days of administration.

[0044] Materials and methods Experimental animals: A chicken farm in Shandong where infectious coryza occurred.

[0045] Test drug: Example 1.

[0046] Experimental method: The diagnosis of infectious coryza in chickens was made based on clinical symptoms, autopsy changes and bacterial isolation and identification. The experimenters randomly divided the sick chickens into two groups in the whole building, and the water line of each group was independently supplied.

[0047] Diagnostic criteria for infectious coryza in chickens: Clinical symptoms: The diseased chickens showed symptoms of nasal mucosal inflammation, runny nose, eyelid edema, difficulty breathing, snoring and strange coughing, sometimes shaking their heads, and green loose stools. Some of the diseased chickens showed the typical unilateral facial edema of the disease, as shown in the following Figure 5 shown.

[0048] Autopsy changes: Cheesy exudates were observed in the sinuses, mucus or pus accumulated in the trachea, and some chickens had mild pneumonia. White pus was seen when the swollen eyelids were pried open. Severely swollen chickens had a yellowish, jelly-like exudate under their facial skin. No food was found in the digestive tract, and the carcasses were extremely emaciated. There were no obvious lesions in the liver, heart, kidneys, bursa of Fabricius, or digestive tract.

[0049] Bacterial isolation and identification: Secretions from the diseased corns, nasal cavity, and infraorbital sinus were collected, smeared, and Gram-stained for microscopic examination. Gram-negative cocci were found, and they were pleomorphic, occasionally fibrillar, and had capsules around the bacteria.

[0050] Pathogen detection: RT-PCR / PCR method is used to detect pathogens.

[0051] The test results are shown in Table 3: Table 3. Pathogen detection results

[0052] Note: “+” means positive; “-” means negative The results showed that the pathogen detection result was HPG-B positive.

[0053] Methods for determining treatment effectiveness: feed intake, egg production rate of laying hens, and morbidity rate (morbidity rate is determined by random sampling).

[0054] The criteria for determining whether a chicken is sick are obvious swelling on the face or a sticky discharge from the nostrils. The same person should be responsible for determining whether a chicken is sick to avoid subjective factors.

[0055] Experimental results: Table 4. The regional clinical effect of Example 1 on infectious coryza in chickens

[0056] Note: The same letters indicate no significant difference in the same column (P≥0.05); different letters indicate significant difference in the same column (P<0.05).

[0057] The results in Table 2 show that the effect of Example 1 is significantly better than that of the negative control group, indicating that after treatment with Example 1, the chickens' appetite increased, clinical symptoms were significantly alleviated, snoring and strange coughing sounds were greatly reduced, and the incidence rate was significantly reduced, which were all significantly improved compared with the control group.

[0058] Compared to the current common clinical approach to preventing and treating infectious rhinitis in chickens, which uses antibiotics but is contraindicated during the laying period for sensitive antibiotics, the present invention uses a pure traditional Chinese medicine preparation instead of antibiotics. This preparation can be used during the laying period, achieving significant improvements compared to the control group. This addresses the lack of available treatment options for infectious rhinitis in laying hens, providing a more effective solution for the prevention and treatment of infectious rhinitis in laying hens.

[0059] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

Claims

1. Application of Mahonia in the preparation of medicine for infectious rhinitis in chickens.

2. The use according to claim 1, characterized in that: Its dosage form is mixture, powder, granule or decoction.

3. A drug for treating infectious rhinitis in chickens, characterized in that: The active ingredient is Mahonia.

4. The drug according to claim 3, characterized in that The active ingredient also contains Nandina domestica.

5. The drug according to claim 3, characterized in that The active ingredients are composed of Mahonia and Nandina domestica.

6. The drug according to claims 3-5, characterized in that Its dosage form is mixture, powder, granule or decoction.

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