Application of an effective extract of a traditional Chinese medicine compound in the preparation of a drug against Eimeria tenella

Abstract

The invention discloses a compound Chinese medicine (referred as modified Gegenqinlian decoction), comprising the following medicinal materials by weight: 30-35% of Magnolia officinalis, 15-20% of Coptis chinensis, 10-15% of Licorice, 15-20% of Puerariae lobata, and 15-20% of Scutellaria baicalensis. The inventor uses the pathogen of Eimeria tenella to prepare a chicken coccidiosis model, and studies the modified Gegenqinlian decoction. The results show that the combined application of the five Chinese medicines has a good preventive and therapeutic effect on chicken coccidiosis, has a significant effect of inhibiting Eimeria tenella, and its insect resistance index is as high as 233.47, and the insect resistance effect is better than the conventional insect resistance drug diclazuril on the market. The present invention is a drug for preventing and treating chicken coccidiosis with good efficacy, no pollution, no residue, improving immunity, meat quality and body weight, and easy use, which provides a new choice for replacing chemical drugs for preventing and treating coccidiosis.

Description

Technical Field

[0001] The invention belongs to the technical field of treatment of avian Eimeria infection, and in particular relates to application of an effective extract of a traditional Chinese medicine compound in preparing a medicine for resisting Eimeria tenella. Background Art

[0002] Chicken coccidiosis is an important and common disease in the poultry industry. Since Eimier discovered chicken coccidiosis in 1870, the harm of chicken coccidiosis has gradually been taken seriously by the world. At present, intensive chicken farms have become the most suitable place for the outbreak of coccidiosis, with an incidence rate of 50%-70%, a mortality rate of 20%-30%, and up to 80% in severe cases. Among various chicken diseases, coccidiosis has the highest incidence rate, which seriously affects the development of the poultry industry, and the decline in egg production often causes greater economic losses than the death of chickens. At present, the control of coccidiosis mainly relies on chemical drugs, but the two major problems of drug resistance and drug residues caused by long-term abuse of chemical drugs are becoming more and more serious. The drug resistance of coccidia and the high incidence and mortality caused by it have always hindered the healthy development of the poultry industry. In recent years, the research on the application of Chinese herbal medicine to prevent and treat chicken coccidiosis has made certain progress, which can not only prevent and treat coccidiosis, but also solve the problem of drug residues in meat and eggs.

[0003] The traditional Chinese medicine formula Gegenqinlian decoction (GQD) is composed of kudzu root, scutellaria, coptis root, and liquorice. Oral administration of kudzu root can inhibit the activation of NF-κB by reducing the expression of Toll-like receptor 4 (TLR4) in the mucosa, downregulating cytokines such as tumor necrosis factor (TNF-α), interleukin (IL)-6, IL-1β, and IL-4, reducing myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels, increasing glutathione synthesis, alleviating oxidative stress in the colon, and relieving the degree of intestinal inflammation. Gegenqinlian decoction may downregulate the production and expression of proinflammatory cytokines and inflammatory mediators through its potential antioxidant effect, inhibiting the NF-κB signaling pathway. GQD can also be used in combination with other drugs. The combination therapy of GQD and anti-mouse PD-1 can downregulate PD-1 and increase the expression of IL-2 and IFN-γ, indicating that the combination therapy can effectively restore T cell function by inhibiting checkpoints. Magnolia officinalis has antibacterial, anti-inflammatory and antioxidant activities, and has certain therapeutic effects on asthma, gastrointestinal diseases and neurological diseases. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an application of an effective extract of a traditional Chinese medicine compound in the preparation of a drug against Eimeria tenella, thereby providing a new approach for treating chicken coccidiosis and avoiding drug resistance and drug residues caused by chemical drug treatment.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] A compound Chinese medicine comprises the following medicinal materials in weight fractions: 30-35% of magnolia bark, 15-20% of coptis root, 10-15% of liquorice root, 15-20% of kudzu root and 15-20% of scutellaria baicalensis.

[0007] The above compound Chinese medicine is used as a feed additive, and the dosage of the feed additive is 500g / ton of feed.

[0008] Application of the traditional Chinese medicine compound in preparing drugs for resisting Eimeria tenella or treating coccidiosis.

[0009] Drugs for fighting against Eimeria tenella or treating coccidiosis include the above-mentioned compound Chinese medicine and chemically synthesized drugs for fighting against chicken coccidiosis.

[0010] The chemically synthesized anti-coccidiosis drugs for chickens are one or more of sulfamethoxazole, sulfaquinoxaline, sulfachloropyrazine, amprolium, dinitolamide, nicarbazine, diclazuril and toltrazuril.

[0011] The above-mentioned drug against Eimeria tenella or for treating coccidiosis is used as a feed additive, and the dosage of the feed additive is 500g / ton of feed.

[0012] Chicken coccidiosis is a common intestinal disease of free-range chickens. The main symptoms of this disease are bloody stool, diarrhea, decreased growth performance and increased feed-to-meat ratio. According to the theory of traditional Chinese veterinary medicine, chicken coccidiosis is a type of disease caused by exogenous coccidiosis, which is caused by coccidiosis consuming blood and damaging body fluid, and damp heat is trapped in the spleen. Therefore, the initial symptoms of sick chickens are less food. They are in low spirits, excrete thin feces, and then there is mucus and blood in the feces. Sick chickens are thirsty and like to drink, are afraid of cold and gather together, and their combs are white, emaciated, and die of exhaustion. Since the clinical symptoms caused by chicken coccidiosis are mainly bloody diarrhea, the treatment is mainly based on insecticide, clearing heat and drying dampness. Based on this, the inventor has developed a compound Chinese medicine (referred to as Jiawei Gegen Qinlian Decoction) by adding flavors based on Gegen Qinlian Decoction, including the following medicinal materials by weight: Magnolia officinalis 30-35%, Coptis chinensis 15-20%, Licorice 10-15%, Pueraria lobata 15-20%, and Scutellaria baicalensis 15-20%. In the prescription: Magnolia officinalis and Pueraria root are heat-clearing and detoxifying drugs, which have the effects of clearing heat and detoxifying, stopping diarrhea and killing insects, and are the main drugs; Coptis chinensis and Scutellaria baicalensis clear heat and dry dampness, thicken the intestines and stop diarrhea, so they are the ministerial drugs; Licorice can reconcile all the drugs and improve the efficacy, and is the guiding drug. The inventor used the pathogen of Eimeria tenella to make a chicken coccidiosis model and studied the modified Pueraria root and Scutellaria baicalensis decoction. The results showed that the combined use of the five traditional Chinese medicines has a good preventive and therapeutic effect on chicken coccidiosis, and has a significant effect of inhibiting Eimeria tenella. Its insect resistance index is as high as 233.47, and its insect resistance effect is better than the conventional anti-insect drug diclazuril on the market.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] (1) The present invention combines the good anti-coccidiosis activity of Magnolia officinalis with the intestinal protection effect of Gegenqinlian decoction, significantly improving the problems of decreased growth performance and animal death caused by coccidiosis.

[0015] (2) The medicinal materials in the present invention are all commonly used traditional medicinal materials, and do not cause any toxic side effects to the chicken itself. It is a drug for preventing and treating chicken coccidiosis, which has good efficacy, is pollution-free, has no residue, improves immunity, meat quality and body weight, and is easy to use. It provides a new choice for replacing chemical drugs for preventing and treating coccidiosis. DETAILED DESCRIPTION

[0016] Preclinical trials

[0017] 1. Material Preparation

[0018] 1.1 Drug preparation

[0019] Modified Gegenqinlian decoction original medicine: 937.5g of Magnolia officinalis, 562.5g of Coptis chinensis, 375g of Licorice, 562.5g of Pueraria root, and 562.5g of Scutellaria baicalensis are prepared in proportion, and the processed modified Gegenqinlian decoction original medicine purchased from the pharmacy is crushed through a 60-mesh sieve as the basic raw material, and then mixed with the powdered antibacterial drug in a mixer according to the weight ratio, and mixed with the feed according to the conventional method. The drugs, dosages, and administration methods used in each embodiment are shown in Table 1.

[0020] Table 1 Drugs and products of Examples 1-4

[0021]

[0022] The nicarbazine used in the examples is a conventional anticoccidial drug nicarbazine premix (Zhejiang Huineng Biotechnology Co., Ltd., 25%).

[0023] 1.2 Chicks: One-day-old laying hens, provided by Beijing Agricultural College, were kept in a special animal house that had been disinfected and free of coccidia; the chicken cages, feed troughs, and water troughs were strictly disinfected, and the chickens were free to eat and drink purified water.

[0024] 1.3 Feed: provided by Da Bei Nong Feed Factory. Example 1 was used as a blank control group without using any anticoccidial drugs.

[0025] 2. Experimental Methods

[0026] 2.1 Trial Grouping

[0027] The experiment selected 2000 chickens from the same batch and randomly divided them into 4 treatments, each with 5 replicates, each with 100 chickens, and the same feeding method was adopted. They were respectively Example 1 Group (basic diet), Example 2 Group (1 ton of basic diet with 3 kg of the original medicine of Gegenqinlian Decoction added), Example 3 Group (1 ton of basic diet with 3 kg of the original medicine of Gegenqinlian Decoction added and 125 g of nicarbazine added) and Example 4 Group (125 g of nicarbazine added to the basic diet).

[0028] 2.2 Feeding and management

[0029] Before the test, the chicken house was cleaned and disinfected. Test period: from 1 day to 28 days old.

[0030] Feed intake and body weight were calculated every 7 days, and diarrhea and bloody stools were recorded every day from the 7th to the 14th day, and every 7 days after the 14th day.

[0031] Immunization procedure: follow the existing immunization procedure of the chicken farm.

[0032] 3. Prevention comparison test

[0033] One-day-old native chickens were randomly picked as the subjects of the preventive comparison test and were randomly divided into 4 groups. After feeding for 7 days, the mental state, feed intake, feces, etc. of the chickens were observed and recorded every day. Finally, the weight gain and feed-to-meat ratio of each group of chicks were calculated. The results are shown in Table 2.

[0034] Table 2 Feed conversion rate FCR of Examples 1-4

[0035]

[0036] As shown in Table 2, the feed-to-meat ratio of Example 2 is 0.09 lower than that of Example 1 (basic diet group), indicating that the original medicine of the modified Gegenqinlian decoction can promote the growth of chicks and does not cause any toxic side effects on the growth of chicks.

[0037] 4. Treatment comparison trials

[0038] 4.1 Test materials

[0039] 4.1.1 Drug preparation

[0040] The preparation method of each group of experimental drugs refers to 1.1, and the dosage and administration method are shown in Table 3.

[0041] The dinitolamide used in the examples is a conventional anticoccidial drug dinitolamide premix (Guangzhou Huixin Animal Pharmaceutical Co., Ltd., 25%), and the diclazuril used is a conventional anticoccidial drug diclazuril premix (Shandong Xundakang Animal Pharmaceutical Co., Ltd., 0.5%).

[0042] 4.1.2 Chicken coccidian oocysts: Sporulated oocysts of Eimeria tenella (donated by China Agricultural University) were first passaged and rejuvenated in two-week-old chickens. The cecum was collected and the oocysts were collected. When the sporulation rate reached more than 80% at 28°C, the oocysts were counted and stored in 2.5% potassium dichromate solution in a 4°C refrigerator. They were used within one month.

[0043] 4.1.3 Chicks: One-day-old chicks are kept in a dedicated animal room that has been disinfected and fumigated in advance to ensure that the animal room, chicken cages, water troughs and feed troughs are thoroughly disinfected and free of coccidia infection. They are fed with feed and water that have been sterilized by high temperature disinfection and are free to eat. One week before the animal coccidiosis model is created, the clinical symptoms of the chickens are carefully observed and their feces are tested for coccidia oocysts. Weak chickens and chickens that may be infected with coccidia are eliminated, and the remaining chickens are kept for later use.

[0044] 4.1.4 Feed: does not contain any anticoccidial drugs or various antibiotics.

[0045] 4.2 Test methods

[0046] Grouping: The remaining 10-day-old chicks were weighed one by one, and the chicks with large weight differences were eliminated. After screening, the chicks with uniform weight were randomly divided into 10 groups and fed the same feeding method.

[0047] Preparation of animal coccidia model: In Example 7-15, chicks were orally administered 2.5*10 sporulated oocysts of Eimeria tenella. 5 Each bird was fed with the corresponding mixed medicine on the same day.

[0048] On the 4th day after modeling, bloody stools began to appear, and the amount of bloody stools was recorded. All chicks were dissected 7.5 days after modeling to observe the degree of cecal lesions. Finally, the relative weight gain and feed conversion rate of each group of chicks and the anticoccidial index (ACI) were calculated. The experimental groups are shown in Table 3.

[0049] Table 3 Formulations and products of Examples 6-15

[0050]

[0051] The morbidity, cure rate and mortality of each group of native chickens were observed and recorded. The efficacy determination method and standard anticoccidial index (ACI) were calculated according to the formula recommended by Merck Company of the United States: ACI = (relative weight gain rate + survival rate) - (oocyst value + lesion value); relative weight gain rate = (weight gain of the test group ÷ weight gain of the blank control group) × 100%; survival rate = (number of surviving chicks in each group ÷ total number of chicks in each group) × 100%. Lesions were scored according to five levels, as shown in Table 4.

[0052] Table 4 Lesion scores

[0053]

[0054] The efficacy is judged as excellent when ACI ≥ 180, good when 160 ≤ ACI ≤ 180, moderate when 140 ≤ ACI ≤ 160, poor when 120 ≤ ACI ≤ 140, and ineffective when ACI ≤ 120. The oocyst value is converted from the number of oocysts per gram (OPG) in the cecal contents.

[0055] 5. Experimental Results

[0056] Example 6: The chicks were not inoculated with sporulated oocysts of Eimeria tenella, and they were in good spirits, ate normally, and had no bloody stools.

[0057] Example 7 On the second day after inoculation with sporulated oocysts of Eimeria tenella, the chicks showed symptoms such as depression, standing still, and loss of appetite; on the fourth day, obvious bloody stools began to appear, but no deaths occurred, and the bloody stools gradually decreased.

[0058] The chickens of Examples 8-15 were autopsied, and the cecum was taken out. The anticoccidial index of each group was calculated according to the lesion score, relative weight gain rate, survival rate and oocyst value. The results are shown in Table 5.

[0059] Table 5 Evaluation of anticoccidial effect

[0060]

[0061] From the data in Table 5, it can be seen that the therapeutic effect of the drug of the present invention on coccidiosis in chicks is significantly better than that of the Western medicine treatment group. It can be seen from Examples 6-11 that the dinitolamide group has no obvious effect alone, and the anticoccidial effect is significantly improved after being used in combination with the present invention; compared with Examples 6-7 and 12-15, it can be seen that in this experiment, at a lower dose equivalent to 125 mg per kilogram of feed in the monomer formula, the modified Gegenqinlian Decoction showed a certain anticoccidial effect. Compared with the control group, the bloody stool discharge of the chickens in the drug administration group was significantly reduced; the lesion score was also significantly reduced compared with the control group. Moreover, the monomer formula can significantly increase the weight of chickens and improve the growth performance of broilers. In addition, the anticoccidial index of dinitolamide and the present invention only reached 233.47 at the highest, showing a high-efficiency anticoccidial effect.

Claims

1. A compound Chinese medicine, characterized in that The invention is composed of the following medicinal materials in the following weight fractions: 30-35% of magnolia bark, 15-20% of coptis root, 10-15% of liquorice root, 15-20% of kudzu root and 15-20% of scutellaria root.

2. Use of the Chinese medicinal compound according to claim 1 in preparing drugs for resisting Eimeria tenella or treating coccidiosis.

3. A drug for treating Eimeria tenella or coccidiosis, characterized in that It comprises the compound Chinese medicine as claimed in claim 1 and a chemically synthesized anti-chicken coccidioides drug.

4. The drug for treating Eimeria tenella or coccidiosis according to claim 3, characterized in that The chemically synthesized anti-chicken coccidiostat is one or more of sulfamethazine, sulfaquinoxaline, sulfachloropyrazine, amprolium, dinitolamide, nicarbazine, diclazuril and toltrazuril.

Images