Anticoccidial plant extract, additive and use thereof

Abstract

This invention relates to the field of feed technology, specifically to an anticoccidial plant extract, additives, and their applications. The anticoccidial plant extract of this invention is an aqueous extract of a combination of plant raw materials, including *Dichroa febrifuga*, *Portulaca oleracea*, *Cirsium japonicum*, *Platycladus orientalis* leaves, *Paeonia lactiflora*, *Astragalus membranaceus*, and licorice. This invention also proposes additives containing this plant extract, including feed additives and drinking water additives. This invention uses natural plant extracts to prevent and treat coccidiosis in poultry, exhibiting green and safe characteristics. Through the synergistic effect of the extracted components, it improves the growth performance of poultry infected with coccidia and reduces the number of coccidia oocysts in feces.

Description

Technical Field

[0001] This invention relates to the field of feed technology, specifically to an anticoccidial plant extract, additive, and its application. Background Technology

[0002] Avian coccidiosis is currently one of the most prevalent parasitic diseases in the global poultry industry. It causes significant economic losses to the poultry industry annually, and its control is becoming increasingly complex. This disease is caused by the apical complex parasite *Eimeria*. *Eimeria* parasites invade the gastrointestinal tract of poultry, damaging intestinal epithelial cells and easily disrupting the mucosal barrier, leading to anemia, emaciation, slow growth, secondary bacterial enteritis, and in severe cases, death.

[0003] Currently, coccidiosis is mainly controlled by administering antibiotics, but improper use of antibiotics can lead to drug resistance in poultry, affect human health, and cause environmental pollution. While antibiotics are the first choice for most commercial broiler farms, drug resistance has become a global problem. With years of continuous use of single drugs, Eimeria strains in multiple regions worldwide have developed resistance to these drugs. To slow the development of resistance, rotation and combination therapy have become standard practices, requiring strict adherence to drug withdrawal periods.

[0004] Live attenuated vaccines are widely used in the prevention and control of coccidia in broilers and laying hens. Live attenuated vaccines induce immunity against specific coccidia strains in poultry by feeding chicks a vaccine mixture (or spraying). While effective, they present challenges such as feeding difficulties, immune failure (due to stress or drug interference), and limited protection against precocious coccidia strains. Subunit vaccines, as a hot research area for next-generation vaccines, offer high safety, but have not yet replaced live vaccines in large-scale production due to high production costs, complex immunization procedures (requiring multiple immunizations or adjuvants), and limited protective efficacy.

[0005] In order to meet the demand for antibiotic-free products in livestock and poultry production, it is necessary to provide an anticoccidial plant extract that is free of drug residues and has proven efficacy. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides an anticoccidial plant extract that not only meets the demand for antibiotic-free products in livestock and poultry production, but also overcomes the drawbacks of inconvenient mixing and operation and difficulty in controlling uniformity due to its good water solubility.

[0007] This invention proposes an anticoccidial plant extract, wherein the anticoccidial plant extract is an aqueous extract of a plant raw material composition, the plant raw material composition including Dichroa febrifuga, Portulaca oleracea, Cirsium japonicum, Platycladus orientalis leaves, Paeonia lactiflora, Astragalus membranaceus and Glycyrrhiza uralensis.

[0008] Optionally, the plant raw material composition includes: 50-100 parts by weight of *Dichroa febrifuga*, 50-100 parts by weight of *Portulaca oleracea*, 50-100 parts by weight of *Cirsium japonicum*, 50-150 parts by weight of *Platycladus orientalis* leaves, 50-100 parts by weight of *Paeonia lactiflora*, 50-100 parts by weight of *Astragalus membranaceus*, and 50-100 parts by weight of licorice; or alternatively, the plant raw material composition includes: 100 parts by weight of *Dichroa febrifuga*, 100 parts by weight of *Portulaca oleracea*, 100 parts by weight of *Cirsium japonicum*, 150 parts by weight of *Platycladus orientalis* leaves, 50 parts by weight of *Paeonia lactiflora*, and *Astragalus membranaceus*. 50 parts by weight of licorice; optionally, the plant raw material composition includes: 50 parts by weight of Dichroa febrifuga, 50 parts by weight of Portulaca oleracea, 50 parts by weight of Cirsium japonicum, 50 parts by weight of Platycladus orientalis leaves, 100 parts by weight of Paeonia lactiflora, 100 parts by weight of Astragalus membranaceus, and 100 parts by weight of licorice; optionally, the plant raw material composition includes: 80 parts by weight of Dichroa febrifuga, 80 parts by weight of Portulaca oleracea, 80 parts by weight of Cirsium japonicum, 100 parts by weight of Platycladus orientalis leaves, 80 parts by weight of Paeonia lactiflora, 80 parts by weight of Astragalus membranaceus, and 80 parts by weight of licorice.

[0009] Optionally, excipients are also added to the anticoccidial plant extract; Optionally, the excipients include one or more of the following: excipients, wetting agents, moisture-proofing agents, flavoring agents, preservatives, and antioxidants; optionally, the excipients are selected from one or more of the following: sugars and water-soluble starches. Optionally, the water-soluble starch is selected from one or more of pregelatinized starch, dextrin, and cyclodextrin; the sugar carrier is selected from sucrose.

[0010] Optionally, the anticoccidial plant extract is prepared using the following steps: Step 1: Weigh the plant raw material composition and extract it with water to obtain an aqueous extract; Step 2: Combine and concentrate the aqueous extracts to obtain a concentrated solution; Step 3: Add a water-soluble carrier to the concentrate, and then dry and pulverize to obtain the anticoccidial plant extract.

[0011] Optionally, in step 1, the mass ratio of the plant material composition to water is 1:10 to 1:12; Optionally, the steam pressure during extraction is 0.2 to 0.4 MPa, the number of extractions is 2 to 4, and the extraction time is 1 to 2 hours each time.

[0012] Optionally, in step 2, the relative density of the concentrate is 1.09 to 1.15; Optionally, the concentration temperature is 45 ~ 55℃ and the pressure is 0.2 ~ 0.4 MPa.

[0013] Optionally, in step 3, the mass ratio of the concentrate to the water-soluble excipient is 1:1.5 to 2.5.

[0014] Optionally, the drying temperature is 55-85℃ and the drying time is 10-24 hours.

[0015] Optionally, the material can be pulverized to pass through an 80-100 mesh sieve.

[0016] This invention proposes a feed additive containing the above-mentioned anticoccidial plant extract; optionally, the amount of feed additive added per ton of feed is 200 to 500 grams.

[0017] This invention proposes a drinking water additive containing the above-mentioned anticoccidial plant extract; optionally, the amount of the drinking water additive added per ton of water is 250 to 500 grams.

[0018] This invention proposes the application of the above-mentioned anticoccidial plant extract in the preparation of drugs, feed additives or drinking water additives for the treatment or prevention of coccidiosis in poultry.

[0019] The technical solution provided by the embodiments of the present invention has the following advantages compared with the prior art: This invention uses natural plants as raw materials to prepare an anticoccidial plant extract, which is green and safe. Through the synergistic effect of each extracted component, it significantly improves the growth performance of poultry infected with coccidia and reduces the number of coccidia oocysts in feces.

[0020] The plant extract for coccidiosis of this invention has good water solubility and can be used as a feed additive or drinking water additive, overcoming the shortcomings of laborious feed mixing and difficulty in controlling uniformity. Detailed Implementation

[0021] To better understand the above-mentioned objectives, features, and advantages of the present invention, the solutions of the present invention will be further described below. It should be noted that, unless otherwise specified, the embodiments of the present invention and the features thereof can be combined with each other.

[0022] Many specific details are set forth in the following description in order to provide a full understanding of the invention, but the invention may also be practiced in other ways different from those described herein; obviously, the embodiments in the specification are only some embodiments of the invention, and not all embodiments.

[0023] The objective of this invention is achieved through the following technical solution: an anticoccidial plant extract, which is an aqueous extract of a plant raw material composition including *Dichroa febrifuga*, *Portulaca oleracea*, *Cirsium japonicum*, *Platycladus orientalis* leaves, *Paeonia lactiflora*, *Astragalus membranaceus*, and licorice. This invention, through research, has discovered that although *Dichroa febrifuga* has been reported to have anticoccidial effects in poultry, its effect when used alone is limited. Therefore, this invention, through in-depth research using various traditional Chinese medicines with water-soluble active ingredients, has obtained a plant extract with significantly enhanced efficacy, significantly superior to the effect of using *Dichroa febrifuga* alone. In particular, the applicant found that the combined use of *Dichroa febrifuga*, *Portulaca oleracea*, *Platycladus orientalis* leaves, and *Cirsium japonicum* can significantly improve the anticoccidial effect in poultry. The addition of *Paeonia lactiflora*, *Astragalus membranaceus*, and licorice further clears heat and dries dampness, replenishes qi and nourishes blood, thereby enhancing the body's resistance and eliminating dampness and killing parasites.

[0024] In one embodiment, the plant material composition includes: 50-100 parts by weight of Dichroa febrifuga, 50-100 parts by weight of Portulaca oleracea, 50-100 parts by weight of Cirsium japonicum, 50-150 parts by weight of Platycladus orientalis leaves, 50-100 parts by weight of Paeonia lactiflora, 50-100 parts by weight of Astragalus membranaceus, and 50-100 parts by weight of Glycyrrhiza uralensis.

[0025] In one embodiment, the plant material composition comprises the following raw materials: 50-100 parts by weight of Dichroa febrifuga, 50-100 parts by weight of Portulaca oleracea, 50-100 parts by weight of Cirsium japonicum, 50-150 parts by weight of Platycladus orientalis leaves, 50-100 parts by weight of Paeonia lactiflora, 50-100 parts by weight of Astragalus membranaceus, and 50-100 parts by weight of Glycyrrhiza uralensis.

[0026] In one embodiment, the plant material composition comprises: 100 parts by weight of *Dichroa febrifuga*, 100 parts by weight of *Portulaca oleracea*, 100 parts by weight of *Cirsium japonicum*, 150 parts by weight of *Platycladus orientalis* leaves, 50 parts by weight of *Paeonia lactiflora*, 50 parts by weight of *Astragalus membranaceus*, and 50 parts by weight of *Glycyrrhiza uralensis*. In one embodiment, the plant material composition consists of the above-mentioned parts by weight of the raw materials.

[0027] In one embodiment, the plant material composition comprises: 50 parts by weight of *Dichroa febrifuga*, 50 parts by weight of *Portulaca oleracea*, 50 parts by weight of *Cirsium japonicum*, 50 parts by weight of *Platycladus orientalis* leaves, 100 parts by weight of *Paeonia lactiflora*, 100 parts by weight of *Astragalus membranaceus*, and 100 parts by weight of *Glycyrrhiza uralensis*. In one embodiment, the plant material composition consists of the above-mentioned parts by weight of the raw materials.

[0028] In one embodiment, the plant material composition comprises: 80 parts by weight of *Dichroa febrifuga*, 80 parts by weight of *Portulaca oleracea*, 80 parts by weight of *Cirsium japonicum*, 100 parts by weight of *Platycladus orientalis* leaves, 80 parts by weight of *Paeonia lactiflora*, 80 parts by weight of *Astragalus membranaceus*, and 80 parts by weight of licorice. In one embodiment, the plant material composition consists of the above-mentioned parts by weight of the raw materials.

[0029] As can be seen from the above implementation scheme, in the plant raw material composition of the present invention, the use of Dichroa febrifuga, Portulaca oleracea, Cirsium japonicum and Platycladus orientalis leaves in a weight ratio of 1:1:1:1.2~1.5, using a large amount of inexpensive herbs—Portulaca oleracea, Cirsium japonicum and Platycladus orientalis leaves—can not only improve the effect against poultry coccidiosis, but also control costs.

[0030] In one embodiment, the anticoccidial plant extract is further supplemented with excipients, including one or more of excipients, wetting agents, moisture-proofing agents, flavoring agents, preservatives, and antioxidants. The excipients are selected from one or more of sugars and water-soluble starches. The carrier is selected from sugars or water-soluble starches; the sugar is sucrose; the water-soluble starch may be one or more of pregelatinized starch, dextrin, and cyclodextrin.

[0031] In one embodiment, the anticoccidial plant extract is prepared using the following steps: Step 1: Weigh the plant raw material composition and extract it with water to obtain an aqueous extract; Step 2: Combine and concentrate the aqueous extracts to obtain a concentrated solution; Step 3: Add a water-soluble carrier to the concentrate, and then dry and pulverize to obtain the anticoccidial plant extract.

[0032] In one embodiment, in step 1, the mass ratio of the plant material composition to water is 1:10 to 1:12.

[0033] In one embodiment, in step 1, the pressure of the water vapor during extraction is 0.2 to 0.4 MPa, the number of extractions is 1 to 4, and the extraction time is 1 to 2 hours each time.

[0034] In one embodiment, in step 2, the relative density of the concentrate is 1.09 to 1.15.

[0035] In one embodiment, in step 2, the concentration temperature is 45 ~ 55°C and the pressure is 0.2 ~ 0.4 MPa.

[0036] In one embodiment, in step 3, the mass ratio of the concentrate to the water-soluble excipient is 1:1.5 to 2.5.

[0037] In one embodiment, in step 3, the drying temperature is 55-85°C, the drying time is 10-24 hours, and the pulverization is passed through an 80-100 mesh sieve.

[0038] The extract prepared by the above method has good solubility. After 1 gram of plant extract is dissolved in 100 mL of water, there is no precipitate, no insoluble particles, and no black charred foreign matter, which can fully meet the requirements for administration via drinking water.

[0039] In one embodiment, the anticoccidial plant extract is prepared using the following steps: 1. Remove impurities from Changshan, Portulaca oleracea, Cirsium japonicum, Platycladus orientalis leaves, Paeonia lactiflora, Astragalus membranaceus, and Glycyrrhiza uralensis. Mix them together, rinse twice with clean water, add 10 to 12 times the amount of water, and extract twice with a steam pressure of 0.2 to 0.4 MPa for 1 to 1.5 hours each time. 2. Combine the extracts and concentrate them at 50℃ and 0.2 ~ 0.4 MPa until the relative density of the concentrate is 1.09 ~ 1.15; 3. Mix the concentrated solution with the water-soluble carrier at a ratio of 1:1.5 to 1:2.5, dry at 75 to 80°C under normal pressure for 10 to 12 hours, pulverize and pass through an 80 to 100 mesh sieve to obtain the anticoccidial plant extract.

[0040] This invention also proposes a feed additive containing the aforementioned anticoccidial plant extract. The amount of the feed additive added per ton of feed can be 200-500 grams.

[0041] This invention also proposes a drinking water additive containing the aforementioned anticoccidial plant extract. The amount of the anticoccidial feed additive added per ton of water can be 250-500 grams.

[0042] Example 1 1. Weigh the following plant raw material composition according to the proportions: 100 parts by weight of Dichroa febrifuga, 100 parts by weight of Portulaca oleracea, 100 parts by weight of Cirsium japonicum, 150 parts by weight of Platycladus orientalis leaves, 50 parts by weight of Paeonia lactiflora, 50 parts by weight of Astragalus membranaceus, and 50 parts by weight of Glycyrrhiza uralensis. Remove impurities, rinse twice with clean water, add 10 times the amount of water, and extract twice for 1 hour each time. 2. Combine the two extracts and concentrate them at 50℃ and 0.2 MPa until the relative density is 1.15. 3. Mix the concentrated liquid with sucrose at a ratio of 1:2.5, dry at 75°C for 12 hours, and pulverize into 80-100 mesh to obtain the anticoccidial plant extract.

[0043] Example 2 1. Weigh the following plant raw material composition according to the proportions: 50 parts by weight of Dichroa febrifuga, 50 parts by weight of Portulaca oleracea, 50 parts by weight of Cirsium japonicum, 50 parts by weight of Platycladus orientalis leaves, 100 parts by weight of Paeonia lactiflora, 100 parts by weight of Astragalus membranaceus, and 100 parts by weight of Glycyrrhiza uralensis. Remove impurities, rinse twice with clean water, add 12 times the amount of water, and extract twice for 1.5 hours each time. 2. Combine the two extracts and concentrate them at 50℃ and 0.3 MPa until the relative density is 1.09. 3. Mix the concentrated liquid with starch dextrin at a ratio of 1:1.5, dry at 80°C for 10 hours, and pulverize into 80-100 mesh to obtain the anticoccidial plant extract.

[0044] Example 3 1. Weigh the following plant raw material composition according to the proportions: 80 parts by weight of Dichroa febrifuga, 80 parts by weight of Portulaca oleracea, 80 parts by weight of Cirsium japonicum, 100 parts by weight of Platycladus orientalis leaves, 80 parts by weight of Paeonia lactiflora, 80 parts by weight of Astragalus membranaceus, and 80 parts by weight of Glycyrrhiza uralensis. Remove impurities, rinse twice with clean water, add 10 times the amount of water, and extract twice for 1.2 hours each time. 2. Combine the two extracts and concentrate them at 50℃ and 0.4 MPa until the relative density is 1.12. 3. Mix the concentrated liquid with a mixture of sucrose and starch dextrin at a ratio of 1:2.5, with the ratio of sucrose to starch dextrin being 1:4; dry at 80℃ for 8 hours, and pulverize into 80-100 mesh to obtain the anticoccidial plant extract.

[0045] Example 4 The anticoccidial plant extracts prepared in Examples 1-3 were tested: 1. Particle size determination Double-layer sieve separation method: Using a 16-mesh analytical sieve and a 30-mesh analytical sieve, the vibrating sieve is horizontally rotated for 3 minutes, with tapping occurring during sieving. The weight of the material remaining on the sieve is then measured. 100% of the material must pass through the 16-mesh analytical sieve, and the percentage of material remaining on the 30-mesh analytical sieve should be ≤10%. From Examples 1-3, one bag of 10 grams is taken from each example.

[0046] The results of particle size determination are shown in Table 1: Table 1: As shown in Table 1, 100% of the crude extracts from natural plants passed through a 16-mesh sieve and 100% passed through a 30-mesh sieve.

[0047] 2. Determination of water solubility of extract Take 1 gram of the anticoccidial plant extract prepared in Examples 1-3, add it to 100 mL of water, stir for 2 min, let stand for 15 min, and observe the solubility. The solution should be slightly turbid with no precipitate, insoluble particles, or black charred foreign matter at the bottom. The test results are shown in Table 2.

[0048] Table 2: The experimental results above show that the prepared anticoccidial plant extract has good water solubility.

[0049] 3. Determination of loss on drying Take 5 grams of each of the crude natural plant extracts prepared in Examples 1-3, spread them evenly in a drying weighing bottle, dry at 103-105℃ for 4 hours, cool in a desiccator, weigh, and calculate the moisture content based on the weight loss. The moisture content should be ≤10%. The results of the drying loss determination are shown in Table 3.

[0050] Table 3: The experimental results above show that the moisture content of the obtained natural plant crude extract powder is all <10%.

[0051] Example 5 1. Treatment experiment on artificially infected chickens with coccidiosis under normal feeding and management conditions.

[0052] Preparation of infected oocysts: Mixed oocysts were collected from the blood feces of chicks raised in chicken farms using conventional methods. They were placed in petri dishes containing 2.5% potassium dichromate solution and cultured in a water bath at 28°C until the sporulation rate reached 90%. The sporulated oocysts were then collected and stored at 4°C for later use.

[0053] Experimental Design Groups: Forty healthy AA commercial broiler chickens, randomly selected at 10 days of age and raised to 14 days of age, were randomly divided into four groups of 10 each: a negative control group, a positive control group, a diclazuril group, and the group described in Example 1. On the second day after grouping, the chickens were administered medication and orally infected with coccidial sporulated oocysts. All chickens were kept in isolation with free access to feed and water. After oocyst infection, the morbidity and mortality of each group were observed and recorded daily, and bloody stools were scored. On the eighth day post-infection, all chickens were weighed, euthanized, and dissected for lesion scoring and oocyst count.

[0054] Negative control group: Not infected with sporulated coccidia oocysts, no medication required; Positive control group: infected with sporulated coccidia oocysts (number of infected oocysts 5 × 10⁶) 4 (each bird), no medication required; Diclazuril group and Example 1 group: infected with sporulated coccidia oocysts (number of infected oocysts 5 × 10⁻⁶) 4 On the same day, administer drinking water containing 1 mg / L diclazuril and 0.05% crude extract from Example 1, twice daily.

[0055] Therapeutic efficacy evaluation indicators: Bloody stool scoring is an indicator for assessing the occurrence and severity of coccidiosis. Stool scores: 0 points, no blood in the stool; 1 point, a small amount of blood in the stool; 2 points, a significant amount of blood in the stool, otherwise normal; 3 points, a significant amount of blood in the stool, the rest being white mucus or 2 / 3 blood in the stool, otherwise normal; 4 points, entirely bloody stool, or 2 / 3 blood in the stool, the rest being white mucus.

[0056] Cecal lesion scoring and lesion value: 0 points, no visible pathological changes in the intestine; 1 point, scattered hemorrhages on the cecal wall, no thickening of the intestinal wall, and normal contents; 2 points, thickening of the cecal wall, with a small amount of bleeding and exudation; 3 points, cecal enlargement, diffuse intestinal bleeding; 4 points, severe intestinal bleeding or presence of intestinal core, thickening and enlargement of the cecal wall.

[0057] Lesion value = average lesion score of the chicken intestines in each group × 10.

[0058] OPG: After the experimental chickens were euthanized, the intestinal contents were collected, mixed, and the number of oocysts per gram of feces (OPG) was counted using the MeMastei method.

[0059] Oocyte ratio: Oocyte ratio = Number of ovulations in each experimental group / Number of ovulations in the positive control group × 100%.

[0060] Oocyte count: When the ovum ratio is 0%~1%, the ovum count is 0; when it is 2%~25%, it is 5; when it is 26%~50%, it is 10; when it is 51%~75%, it is 20; when it is 76%~100%, it is 40.

[0061] Relative weight gain rate (%) = (Average weight gain of experimental group / Average weight gain of control group) × 100% Anticoccidial Index (ACI): Calculated according to Merck's method, ACI = (Relative weight gain + Survival rate) × 100 - (Lesion value + Oocyst value). ACI is a comprehensive indicator for evaluating the efficacy of anticoccidial drugs. The anticoccidial efficacy of a drug is determined based on ACI. An ACI > 180 is considered a highly effective anticoccidial drug; an ACI of 160-180 is considered a moderately effective anticoccidial drug; an ACI of 120-160 is considered a poorly effective anticoccidial drug; and an ACI < 120 is considered an ineffective anticoccidial drug.

[0062] Efficacy evaluation criteria: The effectiveness of anticoccidial drugs in controlling coccidiosis was evaluated based on stool blood scores. A total stool blood score below 5 points was considered highly effective, 6-10 points moderately effective, 11-15 points lowly effective, and above 16 points ineffective.

[0063] The experimental results are shown in Table 4 below: Table 4: As shown in Table 4, the ACI values ​​for the Example 1 group and the diclazuril group were 182.04 and 184.18, respectively, indicating that they are highly effective anticoccidial drugs (ACI > 180). The blood stool scores for the Example 1 group and the diclazuril group were 5 and 4, respectively, indicating that while the Example 1 group and diclazuril could not completely prevent coccidiosis, they could significantly reduce the severity of the disease in chickens. Compared to the positive control group with a survival rate of only 70%, the chicks in the Example 1 group all survived the challenge, achieving a 100% survival rate. The Example 1 group also showed a significant weight gain effect.

[0064] Example 6 Results of comparative trials on the efficacy of treatment for rabbit coccidiosis Experimental animals and grouping Twenty-four 45-day-old White rabbits, weighing 745±115g, were selected for the experiment. These rabbits had previously experienced a severe natural coccidiosis infection (examination of feces by the saturated saline flotation method showed a large number of coccidia oocysts in each rabbit). They were fed in cages that had been sterilized by flame spraying and observed for 3 days before being randomly divided into 3 groups.

[0065] Traditional Chinese medicine control group: Changshan was extracted by water and prepared into plant extracts according to the examples; Group I is Example 1, with materials mixed at a ratio of 1kg / 2000kg; Group II served as the control group for traditional Chinese medicine, which was mixed into feed at a ratio of 1 kg / 2000 kg. Group III consisted of rabbits with a mixture of sulfamethoxypyrimidine and TMP at a ratio of 5:1, mixed with feed at a rate of 1500 mg / kg. Group IV served as the positive control group.

[0066] Eight rabbits were kept in each group, individually in separate cages. After the experiment began, the drug was mixed into the feed in a certain proportion to make pelleted feed. The rabbits ate the feed continuously for 21 days and had free access to water.

[0067] Result Judgment Indicators Clinical health status and necropsy observation: Daily records were kept of the rabbits' appetite, mental state, and fecal condition. If any rabbits died during the experiment, they were immediately dissected for pathological observation. After the experiment, the rabbits were euthanized for further lesion observation.

[0068] Weight gain: Each animal was weighed on days 0, 7, 14, and 21 of the experiment.

[0069] Feed conversion ratio (feed to weight ratio): The calculation formula is: Feed conversion ratio = Average daily feed consumption per group / Average daily weight gain per group.

[0070] Oocyte count (OPG): Feces were collected from each rabbit on days 0, 7, 14, and 21 of the experiment, and the number of oocysts per gram of feces was calculated using the Memaster's method. If the oocyst density was too high, it was serially diluted before counting.

[0071] Drug efficacy assessment: Through clinical and anatomical observations, using OPG, daily weight gain, and feed conversion ratio as indicators, t-tests were performed on the drug group and the positive control group to determine the therapeutic effect of the experimental drug.

[0072] Test results Clinical health status and mortality rate: Rabbits in the positive control group gradually developed decreased appetite, reduced food intake, and lethargy during the experiment, followed by constipation, then diarrhea, and finally, worsening diarrhea (some accompanied by abdominal distension), with even worse appetite leading to complete loss of appetite, scanty feces, emaciation, and symptoms such as screaming and opisthotonus before death. Deaths began on day 6. In the positive control group, a total of 5 rabbits died by day 11, with a mortality rate of 62.5% (5 / 8). By the end of the experiment, the surviving rabbits had rough, emaciated fur and stunted growth. Rabbits in Example 1 and the rabbit ball-strengthening group showed a gradual increase in appetite from day 3 of the experiment. Throughout the experiment, no abnormalities were observed in food intake, feces, or mental state; they grew rapidly, and no deaths occurred.

[0073] Table 5 shows the excretion of coccidia oocysts after drug administration and the analysis of changes in coccidia oocyst excretion.

[0074] Table 5: Note: The unit for counting coccidia is 10. 4 per g.

[0075] Table 5 shows that four fecal examinations were conducted during the experiment. OPG statistics indicated that the excretion of coccidia oocysts in rabbits in Example 1 group, the traditional Chinese medicine control group, and the rabbit coccidia-specific drug group decreased sharply after one week of treatment, with fecal OPG levels as low as 12.8 ± 0.62 × 10⁻⁶. 4 10.71 ± 0.63 × 10⁻⁶ cells / g 4 0.74±0.26×10⁻⁶ cells / g 4 The OPG level was significantly lower than that of the control group rabbits, indicating a better inhibitory effect on OPG expulsion. After one week of continuous treatment, fecal OPG showed a sustained decrease, and in the later stages of the experiment, the vast majority of rabbits did not expel any OPG. At the beginning of the experiment, the fecal OPG in the positive control group reached 18.65 ± 9.40 × 10⁻⁶. 4 The fecal OPG level was 34.86 ± 19.40 × 10⁻⁶ cells / g, while after one week it reached 34.86 ± 19.40 × 10⁻⁶ cells / g. 4 The OPG level decreased in the later stages due to the rabbits' own immunity to coccidia. This indicates that the experimental rabbits were severely infected with coccidia naturally.

[0076] Table 6 shows the daily weight gain after medication.

[0077] Table 6: Note: The values ​​in parentheses are the number of rabbits in each group at the time of weighing; weight unit: g / day / rabbit.

[0078] As shown in Table 6, the average daily weight gain of rabbits in each group during the experiment, from largest to smallest, was as follows: Rabbit Coccidiosis Relief Group (24.0g), Example 1 Group (22.7g), and Traditional Chinese Medicine Control Group (20.4g). Compared with the positive control group (4.8g), the average daily weight gain of each group increased by 400%, 373%, and 325%, respectively. Statistical analysis showed that the differences in average daily weight gain between Example 1 Group, Rabbit Coccidiosis Relief Group, and the positive control group were extremely significant (p < 0.01); however, there was no significant difference between Example 1 Group and Rabbit Coccidiosis Relief Group (p > 0.05). This indicates that the composition obtained in Example 1 and Rabbit Coccidiosis Relief Group can effectively protect rabbits from weight loss caused by coccidiosis infection. During days 8–21 of the experiment, rabbits in the positive control group experienced slower weight gain and weight loss, with an average daily weight loss of 1.6g from days 8 to 14.

[0079] Table 7 shows the feed conversion rate after medication.

[0080] Table 7: Note: The values ​​in parentheses represent the number of rabbits in each group at the time of weighing; "-" indicates negative growth and a negative feed conversion rate.

[0081] As shown in Table 7, the feed conversion ratio (feed to weight ratio) of each group during the entire experiment, from smallest to largest, is as follows: Rabbit Ball Spirit Group 2.83:1, Traditional Chinese Medicine Control Group 2.97:1, Example 1 Group 2.95:1. Compared with the positive control group 7.96:1, the feed conversion ratio of each drug group increased by 181.3%, 168.0%, and 169.8%, respectively.

[0082] Autopsy results: In rabbits that died within 11 days prior to drug administration, the lesions were mainly in the small intestine, showing intestinal bloating, congestion and hemorrhage of the small intestinal wall, thickening of the intestinal wall, narrowing of the intestinal lumen, excessive mucus in the intestinal lumen, catarrhal inflammation of the intestinal mucosa, and loose, unformed feces. Some rabbits showed necrotic lesions in the intestinal wall, and grayish-white nodules on the jejunal mucosa; the urine was yellow with a lot of flocculent material, and the rabbits were extremely emaciated after death. When all rabbits were euthanized and observed 21 days after the experiment, no gross lesions were found in the intestines of any group. However, in the positive control group, yellowish-white nodules were found in the liver and gallbladder, the gallbladder was slightly enlarged, and coccidia oocysts were visible under microscopic examination.

[0083] The above experimental results show that the crude extract prepared in Example 1 safely and significantly inhibits the production and excretion of coccidia oocysts, inhibits the endogenous development of coccidia, reduces intestinal lesions, reduces the mortality rate of young rabbits, and has the effects of increasing weight and improving feed efficiency.

[0084] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the present invention is not to be limited to the embodiments described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. An anticoccidial plant extract, characterized in that, The anticoccidial plant extract is an aqueous extract of a plant raw material composition, which includes Dichroa febrifuga, Portulaca oleracea, Cirsium japonicum, Platycladus orientalis leaves, Paeonia lactiflora, Astragalus membranaceus, and Glycyrrhiza uralensis.

2. The anticoccidial plant extract according to claim 1, characterized in that, The plant raw material composition includes: 50-100 parts by weight of Dichroa febrifuga, 50-100 parts by weight of Portulaca oleracea, 50-100 parts by weight of Cirsium japonicum, 50-150 parts by weight of Platycladus orientalis leaves, 50-100 parts by weight of Paeonia lactiflora, 50-100 parts by weight of Astragalus membranaceus, and 50-100 parts by weight of Glycyrrhiza uralensis. Optionally, the plant material composition includes: 100 parts by weight of Dichroa febrifuga, 100 parts by weight of Portulaca oleracea, 100 parts by weight of Cirsium japonicum, 150 parts by weight of Platycladus orientalis leaves, 50 parts by weight of Paeonia lactiflora, 50 parts by weight of Astragalus membranaceus, and 50 parts by weight of Glycyrrhiza uralensis. Optionally, the plant material composition includes: 50 parts by weight of Dichroa febrifuga, 50 parts by weight of Portulaca oleracea, 50 parts by weight of Cirsium japonicum, 50 parts by weight of Platycladus orientalis leaves, 100 parts by weight of Paeonia lactiflora, 100 parts by weight of Astragalus membranaceus, and 100 parts by weight of Glycyrrhiza uralensis. Optionally, the plant material composition includes: 80 parts by weight of Dichroa febrifuga, 80 parts by weight of Portulaca oleracea, 80 parts by weight of Cirsium japonicum, 100 parts by weight of Platycladus orientalis leaves, 80 parts by weight of Paeonia lactiflora, 80 parts by weight of Astragalus membranaceus, and 80 parts by weight of Glycyrrhiza uralensis.

3. The anticoccidial plant extract according to claim 1, characterized in that, The anticoccidial plant extract also contains excipients; Optionally, the excipients include one or more of the following: excipients, wetting agents, moisture-proofing agents, flavoring agents, preservatives, and antioxidants; Optionally, the excipient is selected from one or more of sugars and water-soluble starches; Optionally, the water-soluble starch is selected from one or more of pregelatinized starch, dextrin, and cyclodextrin; the sugar is selected from sucrose.

4. The anticoccidial plant extract according to claim 1, characterized in that, The anticoccidial plant extract was prepared using the following steps: Step 1: Weigh the plant raw material composition and extract it with water to obtain an aqueous extract; Step 2: Combine and concentrate the aqueous extracts to obtain a concentrated solution; Step 3: Add a water-soluble carrier to the concentrate, and then dry and pulverize to obtain the anticoccidial plant extract.

5. The anticoccidial plant extract according to claim 4, characterized in that, In step 1, the mass ratio of the plant material composition to water is 1:10 to 1:12; Optionally, the steam pressure during extraction is 0.2 to 0.4 MPa, the number of extractions is 1 to 4, and the extraction time is 1 to 2 hours each time.

6. The anticoccidial plant extract according to claim 4, characterized in that, In step 2, the relative density of the concentrate is 1.09 to 1.15; optionally, the concentration temperature is 45 to 55°C and the pressure is 0.2 to 0.4 MPa.

7. The anticoccidial plant extract according to claim 4, characterized in that, In step 3, the mass ratio of the concentrate to the water-soluble excipient is 1:1.5 to 2.5; Optionally, the drying temperature is 55-85°C, and the drying time is 10-24 hours; Optionally, the pulverization is performed by passing the material through an 80-100 mesh sieve.

8. A feed additive, characterized in that, It contains the anticoccidial plant extract as described in any one of claims 1 to 7; optionally, the amount of the feed additive added per ton of feed is 200 to 500 grams.

9. A drinking water additive, characterized in that, It contains the anticoccidial plant extract as described in any one of claims 1 to 7; optionally, the amount of the drinking water additive added per ton of water is 250 to 500 grams.

10. The use of the anticoccidial plant extract as described in any one of claims 1 to 7 in the preparation of a medicament, feed additive, or drinking water additive for the treatment or prevention of coccidiosis in poultry.

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