Application of Isodon japonicus in prevention and treatment of chicken infectious rhinitis

Abstract

The application provides an application of corydalis in preventing and treating chicken infectious rhinitis, and the active ingredient can be composed of corydalis alone, the composition is simple and easy to prepare, the corydalis can be used instead of antibiotics in the egg production period of the chicken, no drug residue is required, the cure rate is high, the corydalis is safe and has no toxic side effects, and the corydalis is convenient to use.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of veterinary medicine, and specifically relates to a traditional Chinese medicine composition for preventing and treating chicken infectious rhinitis with Coptis as a core formula. BACKGROUND

[0002] Chicken infectious rhinitis, an acute respiratory infectious disease commonly seen in poultry farming, is caused by Haemophilus paragallinarum and brings huge economic losses to the breeding industry. Sick chickens will show symptoms such as depression, reduced feed and water intake, respiratory inflammation, and facial swelling, and the egg laying rate of laying hens will decrease significantly, and the growth of broilers will be hindered.

[0003] At present, although sulfonamides have a good inhibitory effect on Haemophilus paragallinarum, the drugs are prone to remain in the chicken body, affecting the feed intake and egg laying rate of the chicken flock, and the use of the drugs during the egg laying period has been prohibited. Due to the advantages of being natural, multi-functional, and having low drug residue requirements, traditional Chinese medicine preparations have become a research hotspot for replacing chemical drugs. However, the existing traditional Chinese medicine preparations for preventing and treating chicken infectious rhinitis have many shortcomings. Some traditional Chinese medicine formulas have a large number of medicinal materials, and small poultry cannot ingest a sufficient dose to exert the drug effect. Some only focus on infectious rhinitis itself and lack effective solutions to complications such as poor growth of broilers and decreased egg laying rate of laying hens. Many traditional Chinese medicine formulas have a large number of components, some formulas contain more than ten medicinal materials, and the components are complex, which results in high cost and poor efficacy. These problems need to be solved to meet the demand of the breeding industry for safe and efficient drugs for preventing and treating chicken infectious rhinitis.

[0004] Coptis is mainly used for the treatment of human diseases in the traditional Chinese medicine field, and has the functions of clearing heat and drying dampness, purging fire and detoxifying, and is commonly used for the treatment of damp-heat diarrhea, jaundice and other diseases. SUMMARY

[0005] The present application aims to provide a traditional Chinese medicine composition for preventing and treating chicken infectious rhinitis with Coptis as the only active ingredient, which can replace antibiotics during the egg laying period of chickens, has the advantages of no drug residue, simple preparation, high cure rate, safety, no toxic side effects, and easy use, and can effectively improve the problems of poor growth of broilers and decreased egg laying rate of laying hens caused by chicken infectious rhinitis. The specific technical solutions are as follows:

[0006] 1. Traditional Chinese medicine composition formula: the traditional Chinese medicine composition of the present application is composed of Coptis or Coptis and Dracaena angustifolia.

[0007] 2. Dosage form: the dosage form is a mixture, a powder, a granule or a decoction.

[0008] 3. Preparation method: the present application further provides a preparation method of the traditional Chinese medicine composition, which is to grind and mix the medicinal materials to prepare a powder, or to extract the medicinal materials with water to prepare a decoction.

[0009] 4. Pharmaceutical use: the application of Mahonia in the preparation of medicines for preventing and treating chicken infectious rhinitis.

[0010] Compared with the prior art, the present application has the following beneficial technical effects:

[0011] The present application selects medicinal materials with antibacterial effect for screening and research, and finds that Mahonia or the combination of Mahonia and Nandina domestica has a significant effect in treating chicken infectious rhinitis, which not only effectively inhibits Haemophilus paragallinarum, but also relieves clinical symptoms, improves the resistance of chicken population, and improves the growth condition and egg production performance of laying hens. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 0 points for the clinical symptoms of chicken infectious rhinitis;

[0013] Figure 2 1 point for the clinical symptoms of chicken infectious rhinitis;

[0014] Figure 3 2 points for the clinical symptoms of chicken infectious rhinitis;

[0015] Figure 4 3 points for the clinical symptoms of chicken infectious rhinitis;

[0016] Figure 5 The chicken farm condition for chicken infectious rhinitis. DETAILED DESCRIPTION

[0017] The specific embodiments of the present application will be further explained and described in combination with specific examples.

[0018] EXAMPLE

[0019] Example 1: Take 1000g of Mahonia, crush, and pass through a 100 mesh screen to obtain.

[0020] Example 2: Take 1000g of Mahonia, add 10 times the amount of water, decoct twice for 1.5 hours each time, combine the filtrate, and concentrate to contain 1g / mL of crude drug to obtain.

[0021] Example 3: Take 500g of Mahonia and 500g of Nandina domestica, add 10 times the amount of water, decoct twice for 1.5 hours each time, combine the filtrate, and concentrate to contain 1g / mL of crude drug to obtain.

[0022] Comparative Example 1: Take 1000g of Nandina domestica, add 10 times the amount of water, decoct twice for 1.5 hours each time, combine the filtrate, and concentrate to contain 1g / mL of crude drug to obtain.

[0023] Comparative Example 2: Take 1000g of Gynostemma pentaphyllum, add 10 times the amount of water, decoct twice for 1.5 hours each time, combine the filtrate, and concentrate to contain 1g / mL of crude drug to obtain.

[0024] Comparative Example 3: 1000g of bitter herbs were boiled twice with 10 times the amount of water for 1.5 hours each time. The filtrates were combined and concentrated to a concentration of 1g / mL of raw herbs.

[0025] Comparative Example 4: 1000g of Acanthopanax senticosus bark, add 10 times the amount of water, decoct twice, 1.5 hours each time, combine the filtrates, and concentrate to a crude drug content of 1g / mL.

[0026] Experimental Example 1

[0027] The in vitro anti-Haemophilus effects of Examples 2, 3, 1, 2, 3, and 4 were observed.

[0028] In this experiment, different concentrations of drugs were mixed and dissolved in liquid culture medium, bacteria were inoculated, and the lowest inhibitory concentration (MIC) of the embodiment was determined by the growth of the bacteria.

[0029] Experimental materials: Example 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4; Haemophilus paragallinarum strain B; TSB medium; diluent; 37℃ incubator.

[0030] Experimental methods:

[0031] Preparation of Haemophilus paragallinarum bacterial culture. Culture medium preparation: Weigh 30g of Tryptic Soy Broth (TSB) medium and 10g of yeast extract, add 945mL of purified water, shake thoroughly to dissolve, autoclave at 121℃ for 15min, cool, add 50mL of inactivated newborn calf serum and 5mL of filtered sterilized 1% NAD solution, shake thoroughly to obtain the culture medium. Pick a single Haemophilus paragallinarum colony and add it to the culture medium, incubate at 37℃ for 24h.

[0032] Preparation of culture medium containing the example: The drug was serially diluted with physiological saline to prepare test solutions of different concentrations. 2.5 mL of each dilution of test solution was added to a test tube containing 2.5 mL of double-concentration TSB culture medium and diluted to 1:256.

[0033] Take 20 μL of bacterial suspension and inoculate it into a test tube containing the culture medium of the example to serve as the experimental group sample.

[0034] The same method was used to inoculate test tubes without the example samples as positive control samples.

[0035] Take two test tubes containing culture medium and treat them with physiological saline instead of the drug to serve as negative control samples.

[0036] The samples from the example group, the positive control group, and the negative control group were placed in an incubator at 37°C and cultured for 24 hours. The results were observed. The positive control group showed obvious turbidity, so the culture was stopped and the results were observed.

[0037] Evaluation method:

[0038] When the positive control tube shows bacterial growth (turbidity) and the negative control tube shows no sterile growth (clearness), the lowest concentration at which no sterile growth occurs in the example group is the MIC of the sample against the test bacteria.

[0039] Experimental results:

[0040] Table 1. Minimum inhibitory concentration (MIC) of different drugs against Haemophilus paragallinarum

[0041]

[0042] The results showed that the negative and positive controls were valid. Examples 2 and 3, and comparative examples 1, 2, 3, and 4 all had in vitro antibacterial effects against Haemophilus paragallinarum. Nandina domestica or Mahonia japonica + Nandina domestica had the best in vitro antibacterial effect.

[0043] Experimental Example 2

[0044] Clinical trials for the prevention and treatment of infectious coryza in chickens

[0045] Cleaning and disinfection of the experimental animal room: Formaldehyde fumigation was used, with 250g of potassium permanganate and 500mL of formaldehyde mixed and fumigated for 24 hours; ventilation was carried out for 24 hours after disinfection.

[0046] SPF chickens were weighed and grouped: 260 SPF chickens over 30 days old were removed, and the remaining 240 SPF chickens were divided into 8 groups of 30 chickens each. The weights of the groups were weighed and adjusted so that the weight difference between the groups was less than 3%, and the data were recorded.

[0047] Preparation of bacterial culture medium

[0048] TSA plates: Weigh 40g of Tryptic Soy Agar (TSA) medium, add 845ml of purified water, shake well and heat until fully dissolved, autoclave at 121℃ for 15min, cool to about 60℃, add 50mL of healthy sterile newborn calf serum, 100mL of yeast extract and 5mL of filtered sterile 1% NAD solution, shake well and pour into sterile Petri dishes to make plates.

[0049] TSB medium: Weigh 30g of Tryptic Soy Broth (TSB) medium and 10g of yeast extract, add 945mL of purified water, shake well to dissolve, autoclave at 121℃ for 15min, cool, add 50ml of inactivated newborn calf serum and 5mL of filtered sterilized 1% NAD solution, shake well to obtain the medium.

[0050] Standards for the Preparation and Control of Highly Toxic Substances

[0051] Take freeze-dried B-type avian bacillus paragallinarum seed, streak it onto a TSA plate, and incubate it at 37°C for 16–24 hours. Pick several typical colonies and inoculate them into 100–200 mL of TSB medium. Incubate at 37°C for 12–24 hours and harvest. Dilute the medium 50 times with sterile physiological saline for challenge and use.

[0052] Storage and usage period: Store below -70℃, and use within 7 days.

[0053] 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 for each group was 1×10⁻⁶. 7 CFU / 0.2 mL / animal, the infection site was the infraorbital sinus, and the blank control group was injected with physiological saline.

[0054] Disease diagnosis criteria: Symptoms are defined as meeting one of the following criteria: ① Nasal discharge ranging from clear to serous-viscous secretions, and tearing; ② Swelling of the infraorbital sinus and surrounding areas on one or both sides of the face. If asymptomatic, a score of 0 is given. If symptoms are present, the severity of clinical symptoms is observed by comparing the affected chickens with the control group. Eye swelling and mucous nasal discharge are scored from mild to severe as 1, 2, and 3 (see [link to relevant documentation]). Figures 1-4 ).

[0055] After the challenge, the corresponding medication was administered via gavage for 7 days to ensure that the amount of medication administered to each group was consistent.

[0056] After the drug was administered, each group was given sufficient food and water.

[0057] Experimental results:

[0058] Table 2. Efficacy of each group in preventing and controlling infectious coryza in chickens

[0059]

[0060] Note: Same letters indicate no significant difference (P≥0.05), different letters indicate significant difference (P<0.05).

[0061] The results showed that after 7 days of drug use, Examples 2 and 3 had the best effect on reducing swelling in chickens caused by Haemophilus paragallinarum infection, especially Example 3, which showed a significant difference compared with the model group. Compared with the solution of the present invention, Comparative Examples 1-4 did not show better effects. It can be seen that drugs with better or worse in vitro antibacterial effects than Example 2 may not necessarily have the same effect as oral treatment of infectious coryza in chickens.

[0062] Experimental Example 3

[0063] The following case studies verify the effectiveness of Example 1 in preventing and treating infectious coryza in chickens.

[0064] To verify the clinical efficacy of this invention against infectious coryza in chickens over a large area, an efficacy verification experiment of Example 1 was conducted in a poultry farm infected with infectious coryza. Because laying hens cannot be treated with sensitive antibiotics, the experiment was divided into Example 1 and a negative control group. The efficacy was compared after 7 days of administration.

[0065] Materials and methods

[0066] Experimental animals: A chicken farm in Shandong Province where infectious coryza was occurring.

[0067] Test drug: Example 1.

[0068] Experimental method: Diagnosis of infectious coryza in chickens was made based on clinical symptoms, necropsy findings, and bacterial isolation and identification. The researchers randomly divided the sick chickens into two groups inside the building, with each group receiving water independently.

[0069] Diagnostic criteria for infectious coryza in chickens:

[0070] Clinical symptoms:

[0071] Sick chickens exhibit symptoms such as nasal mucosal inflammation, runny nose, eyelid edema, difficulty breathing, wheezing, and strange coughing sounds. They may also shake their heads and produce green, watery droppings. Some affected chickens show the typical unilateral facial edema characteristic of this disease. (See attached image) Figure 5 As shown.

[0072] Post-mortem changes:

[0073] Caseous exudate was observed in the sinuses, and mucus or pus accumulated in the trachea. Some chickens had mild pneumonia. White pus drained from the swollen eyelids when the surface was lifted. Chickens with severe facial swelling had a pale yellow gelatinous exudate under the skin of their faces. There was no food accumulation in the digestive tract, and the carcasses were extremely emaciated. No obvious lesions were found in the liver, heart, kidneys, bursa of Fabricius, and digestive tract.

[0074] Bacterial isolation and identification:

[0075] Secretions from the eyes, nasal cavity, and infraorbital sinus of diseased chickens were collected, smeared, and examined under a microscope after Gram staining. Gram-negative cocci were found, which were pleomorphic and occasionally filamentous, with capsules around the bacteria.

[0076] Pathogen detection: RT-PCR / PCR method was used to detect pathogens.

[0077] The test results are shown in Table 3:

[0078] Table 3. Pathogen detection results

[0079]

[0080] Note: "+" indicates a positive result; "-" indicates a negative result.

[0081] The results showed that the pathogen detection was positive for HPG-B type.

[0082] Methods for determining treatment effectiveness: feed intake, egg production rate of laying hens, and morbidity rate (morbidity rate is determined by random sampling).

[0083] The criteria for determining the disease are: obvious swelling of the chicken's face or a viscous fluid discharge from the nostrils. The determination of whether a chicken is sick is performed by the same person to avoid the influence of subjective factors.

[0084] Experimental results:

[0085] Table 4. Regional clinical efficacy of Example 1 against infectious coryza in chickens

[0086]

[0087] Note: 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).

[0088] 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 reduced, wheezing and strange coughing sounds were greatly reduced, and the incidence of disease was significantly reduced, all of which were significantly improved compared with the control group.

[0089] The current clinical approach to preventing and treating infectious coryza in chickens commonly involves the use of antibiotics, but sensitive antibiotics are contraindicated during the egg-laying period. This invention uses a pure traditional Chinese medicine preparation to replace antibiotics, which can be used during the egg-laying period and shows a significant improvement compared to the control group. It fills the gap of having no available treatment for infectious coryza in laying hens during the egg-laying period, providing a more effective solution for the prevention and treatment of infectious coryza in chickens.

[0090] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of 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 protection scope of the present invention.

Claims

1. The application of Mahonia japonica and Nandina domestica in the preparation of drugs for infectious coryza in chickens, characterized in that, The mass ratio of Mahonia japonica to Nandina domestica is 1:

1.

2. The application according to claim 1, characterized in that: Its dosage forms include compound preparations, powders, granules, or decoctions.

3. A drug for treating infectious coryza in chickens, characterized in that, The active ingredients consist of Mahonia japonica and Nandina domestica, with a mass ratio of Mahonia japonica to Nandina domestica of 1:

1.

4. The drug according to claim 3, characterized in that, Its dosage forms include compound preparations, powders, granules, or decoctions.

Images