A traditional Chinese medicine compound with anti-coccidiosis effect and a preparation method and application thereof

Abstract

The application discloses a traditional Chinese medicine compound with anti-coccidiosis effect and a preparation method and application thereof, and relates to the technical field of veterinary drugs. The traditional Chinese medicine compound comprises two or more components in radix sophorae flavescens, cusparia, spilanthes acmella, radix scutellariae and bai-cao-ceng, and the mass ratio is 1-10:1-5:1-10:1-5:1-5. In the traditional Chinese medicine compound, the components are combined together, have the effects of resisting insects, eliminating inflammation, stopping diarrhea and dysentery, stopping bleeding and resisting diseases, and have the advantages of low cost, safety, no drug resistance, environmental protection, no residue and no antibiotics, thereby providing a new idea for an antibiotic-free and green healthy breeding mode. In addition, the traditional Chinese medicine compound is simple to prepare and does not need other adjuvants, and can meet the demand of large-scale use in the breeding industry.

Description

Technical Field

[0001] This invention relates to the field of veterinary drug technology, and more specifically, to a traditional Chinese medicine compound with anticoccidial activity, its preparation method, and its application. Background Technology

[0002] Avian coccidiosis (coccidiosis in chicken) is a common and highly damaging parasitic disease in chickens, an acute epidemic parasitic disease caused by one or more coccidia. Currently, seven pathogens are recognized as causing chicken coccidiosis, with *Eimeria tenella* causing the most significant impact on the poultry industry. Different species of coccidia parasitize different parts of the chicken's intestines, and their pathogenicity varies. *Eimeria tenella*, which parasitizes the cecum, is the most pathogenic. Chickens infected with *Eimeria tenella* experience a severe decline in production performance, reduced feed conversion ratio, and clinical manifestations of bloody diarrhea, which can lead to death in severe cases.

[0003] Coccidiosis in chickens is one of the major diseases seriously threatening the development of the poultry industry, causing economic losses of over US$3 billion annually. Currently, the control of coccidiosis mainly relies on chemical drugs and antibiotics. my country has more than twenty commonly used anticoccidial drugs, but all have shown problems such as drug resistance and drug residues. Traditional Chinese medicine (TCM), due to its lower toxicity and less likelihood of inducing drug resistance, is gradually being applied to research on chicken coccidiosis. Although the disease-fighting effects of TCM products developed from natural plants are difficult to match those of antibiotics and chemically synthesized drugs, TCM greatly reduces the risk of drug resistance, avoids the emergence of superbugs, and my country has abundant and widely used TCM resources. Therefore, developing a new green, efficient, safe, and environmentally friendly anticoccidial TCM compound has significant application value in poultry production. Summary of the Invention

[0004] In order to overcome the shortcomings and deficiencies of the prior art, the purpose of this invention is to provide a traditional Chinese medicine compound with anticoccidial effect, its preparation method and application.

[0005] In this traditional Chinese medicine compound, the mass ratio of Sophora flavescens, Coptis chinensis, Portulaca oleracea, Calcined dragon bone, and scorched herbs is 1-10:1-5:1-10:1-5:1-5. The combined effects of these herbs provide antiparasitic, anti-inflammatory, antidiarrheal, hemostatic, and disease-fighting properties. Furthermore, this compound is cost-effective, safe, does not induce drug resistance, is environmentally friendly, leaves no residue, and contains no antibiotics.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] This invention provides a traditional Chinese medicine compound with anticoccal activity, comprising two or more components from Sophora flavescens, Coptis chinensis, Portulaca oleracea, Fulinggan, and Baicaoshuang, in a mass ratio of 1-10:1-5:1-10:1-5:1-5.

[0008] Furthermore, the aforementioned traditional Chinese medicine compound is any one of the following:

[0009] (1) It includes Sophora flavescens, Coptis chinensis, Portulaca oleracea, Fulinggan and Baicaoshuang, in a mass ratio of 1-10:1-5:1-10:1-5:1-5 (preferably 1:1:1:1:1);

[0010] (2) It includes horsetail and purslane, with a mass ratio of 1 to 5: 1 to 10 (preferably 1: 1);

[0011] (3) Includes Sophora flavescens and Fulinggan; the mass ratio is 1-10:1-5 (preferably 1:1);

[0012] (4) It includes Sophora flavescens, Coptis chinensis, Portulaca oleracea and Fulinggan, in a mass ratio of 1-10:1-5:1-10:1-5 (preferably 1:1:1:1);

[0013] (5) It includes Sophora flavescens and Coptis chinensis in a mass ratio of 1-10:1-5 (preferably 1:1).

[0014] Among them, Sophora flavescens has the effects of clearing heat and drying dampness, killing parasites and removing accumulations, and promoting diuresis. It can also promote the function of immune cells and the production of various cytokines to enhance non-specific immune function and achieve the effect of anti-coccidia. Coptis chinensis has the effects of treating damp-heat diarrhea, jaundice, and feverish irritability. It is currently used for enteritis, infectious hepatitis, colds, carbuncles, boils, and conjunctivitis. Purslane has the effects of killing parasites, quenching thirst, clearing heat and detoxifying. It is mainly used to treat dysentery, leukorrhea, and enteritis. It can be used to prevent coccidiosis in chickens and stop diarrhea. Fulinggan has the effects of warming the middle and drying dampness, stopping vomiting and bleeding. Baicaoshuang has the effect of stopping bleeding and can help improve hemoptysis and hematemesis. The combination of these Chinese medicines has the effects of antiparasitic and anti-inflammatory, anti-dysentery and anti-diarrheal, hemostatic and anti-disease. It also has the advantages of low cost, safety without drug resistance, environmental protection without residue, and no antibiotics.

[0015] Through experimentation, the optimal formulation was determined to be a decoction of Sophora flavescens, Coptis chinensis, Portulaca oleracea, Fulinggan (a type of medicinal herb), and Baicaoshuang (another medicinal herb) in a mass ratio of 1:1:1:1:1, with the dosage form being water decoction.

[0016] The effects of the above-mentioned traditional Chinese medicine compound formula or its components on low-dose coccidiosis infection in Lingnan Kuai Da Huang chickens were tested. The results showed that the anticoccidial effect of the five components of the traditional Chinese medicine compound in a 1:1:1:1:1 ratio was the best, followed by the formula of Coptis chinensis and Portulaca oleracea in a 1:1 ratio. The single-herb formulas were less effective than the multi-herb formulas. In addition, the decoction drinking water test group reduced bloody stools caused by coccidiosis, oocyst counts, and intestinal lesions compared with the control group that did not receive any medication. At the same time, it increased the weight of chickens. The anticoccidial index was on par with the control group of chemically synthesized anticoccidial drugs. The final conclusion is that the traditional Chinese medicine compound formula with five components in a 1:1:1:1:1 ratio is the most effective.

[0017] The above-mentioned optimal Chinese herbal compound was used to conduct a high-dose coccidiosis infection effect test on fast-growing white-feathered broilers. The results showed that the Chinese herbal compound decoction drinking water experimental group was better than the control group without medication in terms of mortality, relative weight gain, feed conversion rate, coccidiosis bloody stool, intestinal lesion score and oocyst reduction rate. The anticoccidial index was slightly lower than the control group of chemically synthesized anticoccidial drugs.

[0018] This traditional Chinese medicine compound does not contain antibiotics, so it will not lead to drug resistance, will not expose a large number of livestock and poultry or even humans to the risk of infection caused by drug-resistant strains, and will not pose a threat to food safety. Therefore, this traditional Chinese medicine compound can provide new ideas for antibiotic-free, green and healthy breeding models.

[0019] Preferably, the dosage form of the traditional Chinese medicine compound is a dosage form that can be used for animal drugs, such as powder, tablet, capsule, or decoction.

[0020] In terms of dosage forms for animal medication, the most common methods are feed mixing and drinking water administration. Medications are often added as powders or decoctions to feed or water containers for direct consumption by animals. Powders are primarily used as feed additives, while decoctions are mainly used for prevention and treatment in clinical settings.

[0021] When the traditional Chinese medicine compound is a decoction, its preparation method is to weigh each part of the traditional Chinese medicine raw materials, crush and soak them, extract them by ultrasonic extraction in a water bath, centrifuge them, and then concentrate them by rotary evaporation, and finally make up the volume to obtain the final product.

[0022] The above-mentioned decoction preparation process is simple, requires low-cost raw materials, and does not contain hormones or antibiotics. It conforms to the future industry trend of reducing and replacing antibiotics and environmental protection, and has significant practical significance for green and healthy breeding models.

[0023] Preferably, the Chinese herbal raw materials are pulverized and then soaked in 5 to 20 times their volume of water for 30 to 120 minutes; more preferably, they are soaked in 10 times their volume of water for 30 minutes.

[0024] Preferably, the water bath ultrasound is performed at 50-60°C for 30-60 minutes; more preferably, it is performed at 50°C for 45 minutes.

[0025] Preferably, the extraction time is 60 to 120 minutes; more preferably, it is 120 minutes.

[0026] Preferably, the rotary evaporation process is carried out according to conventional procedures to prepare a concentration of 0.4–2 g / mL.

[0027] This invention also seeks protection for the use of the above-mentioned traditional Chinese medicine compound in the preparation of feed or feed additives or drinking water additives.

[0028] Preferably, the amount of the traditional Chinese medicine compound added is 0.05% to 0.2% (preferably 0.2%) of the drinking water volume;

[0029] Specifically, the traditional Chinese medicine compound can fight coccidiosis, prevent and / or treat coccidiosis in chickens.

[0030] Furthermore, the anticoccidial agent is an anticoccidial agent against Eimeria tenella.

[0031] Furthermore, the aforementioned chicken coccidiosis is caused by Eimeria tenella.

[0032] The present invention has the following advantages and effects compared with the prior art:

[0033] The traditional Chinese medicine compound of this invention, when combined, possesses antiparasitic, anti-inflammatory, antidiarrheal, hemostatic, and disease-fighting effects. It also boasts advantages such as low cost, safety with no drug resistance, environmental friendliness with no residue, and antibiotic-free composition, providing a new approach for antibiotic-free, green, and healthy aquaculture models. Furthermore, the preparation of this traditional Chinese medicine compound is simple, requiring no other adjuvants, and can meet the needs of large-scale use in the aquaculture industry. Attached Figure Description

[0034] Figure 1 This refers to the bloody stool condition of chickens in groups S1-S8 of Example 1.

[0035] Figure 2 This refers to the bloody stool condition of chickens in groups S9-S16 of Example 1.

[0036] Figure 3 This refers to the bloody stool condition of chickens in groups S17-S18 of Example 1.

[0037] Figure 4 These are the cecal lesion results of chickens in groups S1-S6 of Example 1.

[0038] Figure 5 The results are the cecal lesions of chickens in groups S7-S14 of Example 1.

[0039] Figure 6 The results are the cecal lesions of chickens in groups S15-S18 of Example 1.

[0040] Figure 7 This refers to the bloody stool condition of broiler chickens in groups SⅠ-SⅣ in Example 2.

[0041] Figure 8 This is the result of cecal lesions in broiler chickens in groups SⅠ-SⅣ of Example 2. Detailed Implementation

[0042] The present invention will be further described in detail below with reference to embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto. Unless otherwise stated, the raw materials used in the embodiments of the present invention are conventionally purchased raw materials.

[0043] Example 1: Effect of a traditional Chinese medicine compound on low-dose coccidiosis infection in Lingnan fast-growing yellow chickens.

[0044] 1. Preparation of experimental materials

[0045] Traditional Chinese Medicine Compound 1: Take 10g each of the following herbs (Sophora flavescens, Coptis chinensis, Portulaca oleracea, Calcined Osmanthus fragrans, and Calcium affine) in a mass ratio of 1:1:1:1:1, totaling 50g. Crush the herbs and extract using a decoction. Soak in 10 times the volume of water for 30 minutes (500mL), then sonicate at 50℃ for 45 minutes. After two hours of extraction, mix well, collect the supernatant, centrifuge at 2500rpm for 5 minutes, and concentrate by rotary evaporation. Repeat the ultrasonic extraction process with 10 times the volume of water, and mix the supernatants from both extractions. Perform rotary evaporation on the above extract to obtain a final extract concentration of 1g / mL in 50mL.

[0046] The traditional Chinese medicine formula was prepared by combining the components of traditional Chinese medicine compound formula 1 into 14 groups, either as single herbs or as separate formulas. 10g of each single herb was weighed and prepared simultaneously according to the above preparation method, finally obtaining 50mL of extract with a concentration of 0.2g to 0.8g / mL.

[0047] Conventional anticoccidial drugs: Three-character coccidiostat powder (30% sulfachlorpromazine sodium) was provided by Guangzhou Huixin Biotechnology Co., Ltd.; Baiqiuqing (2.5% toltrazuril) was produced by Bayer AG, Germany.

[0048] 2. Experimental animals and feed

[0049] One-day-old Lingnan fast-growing yellow chickens were purchased from Zhiwei Livestock Co., Ltd.; 161Q complete broiler feed, which does not contain any anticoccidial drugs or antibiotics, was purchased from Baiyun District Experimental Feed Factory.

[0050] 3. Test methods and judgment criteria

[0051] 3.1 Experimental Design and Grouping

[0052] Chicks were raised to 12 days of age. After weighing each chick, weak or overweight chicks were removed, and healthy chicks with an individual weight difference within 10g were selected and randomly divided into 18 groups of 15 chicks each. Groups S1-S14 were treated with different formulations of traditional Chinese medicine to induce parasites, while groups S15 and S16 were control groups for two sensitive drugs. A control group (S17) without treatment and a control group (S18) without treatment and no treatment were also established. The traditional Chinese medicine compound treatment groups (S1-S14) were administered the medicine via drinking water starting at 12 days of age, while the sensitive drug groups (S15 and S16) were administered the medicine via drinking water at 14 days of age. At 14 days of age, each chick in the S1-S16 drug treatment groups and the control group (S17) was orally inoculated with 1.0 × 10¹² ppm. 4 Sporulated oocysts of Eimeria tenella GD strain were collected. All experimental chickens had free access to feed and water. They were euthanized on day 7 post-infection for lesion assessment (see Table 1 for details). The sporulated oocysts of Eimeria tenella GD strain (i.e., Eimeria tenella Guangdong strain) were preserved by the Animal Health Institute of Guangdong Academy of Agricultural Sciences.

[0053] Table 1 Experimental Design Grouping

[0054]

[0055] Note: A = Sophora flavescens, B = Coptis chinensis, C = Portulaca oleracea, D = Fulonggan, E = Baicaoshuang.

[0056] 3.2 Criteria for assessing the anticoccidial index

[0057] 3.3.1 Score for bloody stool

[0058] Fecal scoring was performed according to the method of Suo Xun (1997). On the fifth day after infection, the proportion of bloody stool was observed, photographed and recorded, and bloody stool was scored: For Eimeria tenella infection, within a given time range of 12-24 hours, 0 points indicated 100% bloodless stool, +1 point indicated 25% bloody stool, +2 points indicated 50% bloody stool, +3 points indicated 75% bloody stool, and +4 points indicated 100% bloody stool.

[0059] 3.3.2 Relative weight gain rate

[0060] At the beginning and end of the experiment, the weight of the chickens was measured, and the average weight gain and relative weight gain rate were calculated. Relative weight gain rate = (weight gain rate of the experimental group / weight gain rate of the experimental group without medication or parasite treatment) × 100%.

[0061] 3.3.3 Survival rate

[0062] Record the number of dead chickens in each group, perform necropsy to determine the cause of death, and calculate the survival rate. Survival rate = (number of surviving chickens at the end of the experiment / number of chickens in the experimental group) × 100%.

[0063] 3.3.4 Lesion Value

[0064] Chickens were slaughtered on the 7th day after infection. The intestinal lesion score of each chicken was calculated according to the lesion scoring method designed by Johnson and Reid (1970), and the lesion score was converted into a lesion value.

[0065] Lesion scoring: (When lesions on both sides of the cecum are inconsistent, the more severe side shall be used as the score):

[0066] 0 points, no visible lesions;

[0067] 1 point, the cecal wall has a few scattered petechiae, the intestinal wall is not thickened, and the contents are normal;

[0068] 2 points, numerous lesions, obvious blood in the cecal contents, slightly thickened cecal wall, normal contents;

[0069] 3 points, the cecum contains a large amount of blood or a cecal core (blood clots or grayish-white, cheese-like banana-shaped masses), the cecal wall is significantly thickened, and the cecum contains little feces;

[0070] 4 points. Chickens that die from coccidiosis are also given 4 points, as their cecum is swollen due to being filled with a large amount of blood or intestinal contents, and the intestinal contents may or may not contain fecal matter.

[0071] Lesion score (0-40) = Average lesion score (0-4) of each experimental group × 10.

[0072] The reduction rate of lesion score (RLS) = (average lesion score of the control group without drug treatment - average lesion score of the drug treatment group) / average lesion score of the control group without drug treatment * 100%.

[0073] The lesion score reduction rate (RLS) evaluation criteria are as follows: RLS <= 30% in the treatment group is considered to be drug-resistant, 31-49% is considered to be partially drug-resistant, and >= 50% is considered to be sensitive.

[0074] 3.3.5 Oocyte count and relative ovum yield (ROP)

[0075] The fecal oocyst count was performed using the McMaster counting method to determine the number of oocysts per gram of feces (OPG), and the oocyst value was calculated based on Table 2.

[0076] Relative oocyst yield (ROP) = (average oocyst yield in the drug-treated group / average oocyst yield in the untreated control group) × 100%. ROP ≥ 15% indicates drug resistance; ROP < 15% indicates no drug resistance.

[0077] Table 2. Conversion between number of ovules and ovule value

[0078]

[0079] 3.3.6 Anticoccidial Index (ACI)

[0080] According to the calculation formula of Merck & Co., Inc. in the United States, that is:

[0081] ACI is calculated as follows: ACI = (relative weight gain rate + survival rate) × 100 - (lesion value + oocyst value).

[0082] Pharmacodynamic judgment criteria: ACI > 180 belongs to highly effective anticoccidial drugs; 160 < ACI < 180 belongs to moderately effective anticoccidial drugs; 120 < ACI < 160 belongs to low - effective anticoccidial drugs; ACI < 120 is ineffective against coccidia.

[0083] 4 Test results

[0084] 4.1 Clinical symptoms

[0085] After the challenged chickens were infected with sporulated oocysts, they gradually showed reduced feed intake and poor spirit. On the 5th day after infection, some drug - treated groups and the non - drug - treated challenged control group had varying degrees of bloody feces excretion, reduced water intake, which was more severe on the 5th and 6th days. When sacrificed and dissected on the 7th day after challenge, cecal lesions, varying degrees of bleeding or blood clots were observed, and no lesions were observed in other organs; the non - drug - treated and non - challenged control group had no bloody feces, and the feed intake and water intake were normal.

[0086] 4.2 Results of bloody feces scoring and cecal lesions

[0087] Based on the observation results of bloody feces on the 5th day after challenge, the bloody feces conditions of the drug - treated challenged groups S5, S6, and S7 showed +1 point; the bloody feces condition of the non - drug - treated challenged control group S17 showed +2 points; the bloody feces condition of other drug - treated groups and the non - drug - treated and non - challenged control group S18 showed 0 point. The results of bloody feces are shown in Figures 1-3 , and the results of cecal lesions are shown in Figures 4-6 .

[0088] 4.3 Results of drug sensitivity test

[0089] When the water extracts of different formulations of traditional Chinese medicine compound 1 were administered by drinking water at a dose of 2000 ppm and used to infect with a low dose of Eimeria tenella GD strain, the results showed that: the ACI index of group S14 performed the best (179.68), and the relative oocyst production (ROP) value was the lowest (36.23%), and the reduction rate of lesion score (RLS) value was the highest (52.78%), showing a moderately effective anticoccidial effect; the ACI index of group S9 (173.70) was the second, followed by group S8 (166.36), group S13 (166.06), group S6 (161.26), and group S3 (161.11), all showing a moderately effective anticoccidial effect. For other groups, ACI was lower than 160, and both ROP and RLS were lower than the sensitive values, being judged as low - effective or ineffective. The sensitive drug control groups S15 and S16 showed significant improvement in all indicators and had a moderately high - potency anticoccidial effect. The results are shown in Table 3 for details.

[0090] Table 3. Results of drug susceptibility testing

[0091]

[0092]

[0093] Note: A = Sophora flavescens, B = Coptis chinensis, C = Portulaca oleracea, D = Fulonggan, E = Baicaoshuang.

[0094] 5. Conclusion

[0095] 5.1 The control group without drug administration and the control group without drug administration and without drug administration were established in this experiment; the sensitive drug control group was sensitive to the oocyst yield and lesion score of this strain, and could achieve a medium-to-high efficacy of anticoccidial treatment.

[0096] 5.2 The drug in this study was subjected to anticoccidial animal drug sensitivity tests at the recommended dosage concentration. The results showed that the traditional Chinese medicine compound S14 group (Sophora flavescens: Coptis chinensis: Portulaca oleracea: Fulinggan: Baicaoshuang = 1:1:1:1:1) showed the best anticoccidial effect against Eimeria tenella GD strain at a drinking water dose of 2000 ppm, with the best ROP and RLS values. The next best anticoccidial effect was shown in the traditional Chinese medicine decomposed formula S9 group (Coptis chinensis: Portulaca oleracea = 1:1), followed by S8, S13, S6, and S3 groups, all showing moderate anticoccidial effects. The other groups were judged to be ineffective or have an ACI below 160 and ROP and RLS below the sensitivity value.

[0097] Example 2: Effect of a traditional Chinese medicine compound on high-dose coccidiosis infection in fast-growing white-feathered broiler chickens.

[0098] 1. Preparation of experimental materials

[0099] Traditional Chinese Medicine Compound 1: 50g of each of the following ingredients were weighed in a mass ratio of 1:1:1:1:1: 50g of each. The ingredients were pulverized and extracted using a water decoction. The mixture was soaked in 10 times its volume of water for 30 minutes (500mL), then sonicated in a 50℃ water bath for 45 minutes. After two hours of extraction, the mixture was stirred, and the supernatant was centrifuged at 2500rpm for 5 minutes. The supernatant was then concentrated by rotary evaporation. The precipitate after centrifugation was treated with 10 times its volume of water and sonicated once more. The supernatants from both extractions were mixed. The extract was then subjected to rotary evaporation to obtain a final extract concentration of 1g / mL in 50mL.

[0100] Conventional anticoccidial drugs: Three-character coccidial powder (30% sulfachlorpromazine sodium) is provided by Guangzhou Huixin Biotechnology Co., Ltd.

[0101] 2. Experimental animals and feed

[0102] One-day-old fast-growing white-feathered broiler chickens (also known as: 818 broiler hybrid chickens) were purchased from Wens Foodstuff Group's Sanhuang Farm; 161Q complete broiler feed, which does not contain any anticoccidial drugs or antibiotics, was purchased from Wens Foodstuff Group's experimental feed plant.

[0103] 3. Test methods and judgment criteria

[0104] Chicks were raised to 12 days of age. After weighing each chick, weak or overweight chicks were removed, and healthy chicks with an individual weight difference within 10g were selected and randomly divided into 4 groups of 20 chicks each. Group SⅠ was treated with a traditional Chinese medicine compound 1 for worm treatment, Group SⅡ was the control group for conventional drugs, and there were also a control group (SⅢ) without drug treatment and a control group (SⅣ) without drug treatment or worm treatment (i.e., blank control group SⅣ). Drug treatment began in all experimental groups at 12 days of age. Group SⅠ received the drug through drinking water throughout the day, while Group SⅡ received the conventional drug through drinking water according to the instructions. At 14 days of age, each chick in all experimental groups and the control group without drug treatment was orally inoculated with 1.0 × 10¹² gram of traditional Chinese medicine compound ... 5 The tender Eimeria coccidia sporulated oocysts were observed. All chickens in the experimental groups had free access to feed and water until the end of the experiment on day 7, as detailed in Table 4.

[0105] Table 4 Experimental Design Grouping

[0106]

[0107] The test indicators were determined as follows:

[0108] (1) Score for bloody stool

[0109] Fecal scoring was performed according to the method of Suo Xun (1997). On the fifth day after parasite treatment, the proportion of bloody stool was observed, photographed and recorded, and bloody stool was scored: 0 points indicated 100% bloodless stool, +1 point indicated 25% bloody stool, +2 points indicated 50% bloody stool, +3 points indicated 75% bloody stool, and +4 points indicated 100% bloody stool.

[0110] (2) Relative weight gain rate

[0111] At the beginning and end of the experiment, the weight of the chickens was measured, and the average weight gain and relative weight gain rate were calculated. Relative weight gain rate = (weight gain rate of the experimental group / weight gain rate of the experimental group without medication or parasite treatment) × 100%.

[0112] (3) Survival rate

[0113] Record the number of dead chickens in each group, perform necropsy to determine the cause of death, and calculate the survival rate. Survival rate = (number of surviving chickens at the end of the experiment / number of chickens in the experimental group) × 100%.

[0114] (4) Pathological value

[0115] Chickens were sacrificed on the 7th day after infection. According to the lesion scoring method designed by Johnson and Reid (1970), the intestinal lesions of each chicken were scored, and the lesion scores were converted into lesion values.

[0116] Lesion score: (When the lesions in the two ceca are inconsistent, the more severe side is taken as the standard):

[0117] 0 points, no gross lesions;

[0118] 1 point, there are a few scattered petechiae on the cecal wall, the intestinal wall is not thickened, and the contents are normal;

[0119] 2 points, the number of lesions is more, the cecal contents are significantly bloody, the cecal wall is slightly thickened, and the contents are normal;

[0120] 3 points, there is a large amount of blood in the cecum or there is a cecal core (blood clot or grayish-white cheesy banana-shaped mass), the cecal wall is significantly thickened, and the feces content in the cecum is less;

[0121] 4 points, the cecum is enlarged due to being filled with a large amount of blood or cecal core, the cecal core contains fecal residue or not, and chickens that died of coccidiosis are also scored 4 points.

[0122] Lesion value (0 - 40) = average lesion score (0 - 4) of each experimental group × 10.

[0123] Reduction rate of lesion score (RLS) = (average lesion score of the non-drug challenged control group - average lesion score of the drug-administered challenged group) / average lesion score of the non-drug challenged control group * 100%.

[0124] Judgment criteria for the reduction rate of lesion score (RLS): When RLS <= 30% in the drug-treated group, it is judged to have drug resistance; when 31 - 49%, it has partial drug resistance; when >= 50, it is sensitive.

[0125] (5) Oocyst value

[0126] The fecal oocyst count was performed using the McMaster counting method to obtain the number of oocysts per gram of feces (OPG) in each group, and the oocyst value was obtained according to Table 2.

[0127] (6) Anti-coccidial index (ACI)

[0128] According to the calculation formula of Merck Company in the United States, that is:

[0129] ACI = (relative weight gain rate + survival rate) × 100 - (lesion value + oocyst value) is calculated.

[0130] Pharmacodynamic judgment criteria: ACI > 180 belongs to highly effective anti-coccidial drugs; 160 < ACI < 180 belongs to medium-effective anti-coccidial drugs; 120 < ACI < 160 belongs to low-effective anti-coccidial drugs; ACI < 120 is ineffective against coccidia.

[0131] 4. Test Results

[0132] 4.1 Clinical symptoms

[0133] After infection with sporulated oocysts, chickens in the challenge test gradually showed symptoms such as reduced feed intake and decreased mental state. On the 5th day post-infection, both the drug-treated group and the control group without treatment showed varying degrees of bloody feces and reduced water intake, which became more severe on the 5th and 6th days. On the 7th day after treatment, the chickens were euthanized and autopsied, revealing cecal lesions with varying degrees of hemorrhage or blood clots. No lesions were observed in other organs. The control group without treatment did not have bloody feces and had normal feed and water intake.

[0134] 4.2 Stool hemorrhage score and cecal lesion results

[0135] On the fifth day after the parasite treatment, the results of stool blood observation showed that the compound experimental group (SⅠ) and the conventional drug control group (SⅡ) showed a score of +1 for stool blood; the no-drug parasite treatment group (SⅢ) showed a score of +3 for stool blood; and the blank control group (SⅣ) showed a score of 0 for stool blood. The stool blood results are shown in the attached table. Figure 7 The results of the cecal lesions are shown in Figure 8 .

[0136] 4.3 Results of drug sensitivity test

[0137] The anticoccidial index (ACI) of each group was calculated, and the results are shown in Table 5. Among the tested drugs, the ACI of Compound 1 (SⅠ) at 2000 ppm drinking water was 140.30, slightly lower than that of the conventional drug control group (SⅡ). Furthermore, its relative weight gain, survival rate, and oocyst count were all superior to those of the untreated control group (SⅢ). This indicates that Compound 1 has a good anti-Eimeria tenella effect under high-dose Eimeria tenella infection, but its anticoccidial effect is slightly lower than that of the conventional anticoccidial drug, Three-Character Coccidia Powder.

[0138] Table 5. Results of drug susceptibility testing

[0139]

[0140] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A traditional Chinese medicine compound with anticoccidial activity, characterized in that: It is made from Sophora flavescens, Coptis chinensis, Portulaca oleracea, Fulinggan (a type of medicinal herb), and Baicaoshuang (another medicinal herb), in a mass ratio of 1:1:1:1:

1.

2. The traditional Chinese medicine compound according to claim 1, characterized in that: The dosage form of the traditional Chinese medicine compound is powder, tablet, capsule or decoction.

3. The method for preparing the traditional Chinese medicine compound with anticoccidial activity according to any one of claims 1 to 2, characterized in that: When the dosage form of the traditional Chinese medicine compound is a decoction, its preparation method is to weigh each part of the traditional Chinese medicine raw materials, crush and soak them, extract them by ultrasonic extraction in a water bath, centrifuge them, and then concentrate them by rotary evaporation, and finally make up the volume to obtain the final product.

4. The preparation method according to claim 3, characterized in that: After the Chinese herbal raw materials are pulverized, they are soaked in 5 to 20 times their volume of water for 30 to 120 minutes. The conditions for the water bath ultrasound are: ultrasound at 50-60°C for 30-60 minutes. The extraction time is 60–120 min; The rotary evaporation process is carried out according to conventional procedures, and the concentration is prepared to 0.4–2 g / mL.

5. The use of the traditional Chinese medicine compound with anticoccidioidomy effect as described in any one of claims 1 to 2 in the preparation of a drug for at least one of anticoccidioidomy, prevention of coccidiosis in chickens, and treatment of coccidiosis in chickens.

6. The application according to claim 5, characterized in that: The anticoccidial agent is effective against Eimeria tenella; The coccidiosis in chickens mentioned refers to coccidiosis caused by Eimeria tenella.

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