Application of biscoumarin to resistance to eimeria infection

Abstract

The invention discloses application of dicoumarin to resistance to eimeria infection, and belongs to the technical field of application of dicoumarin. The invention not only discloses that the biscoumarin can effectively inhibit the reproduction of the eimeria on an in-vitro eimeria cell culture model, but also discloses application of the biscoumarin in preparation of the eimeria infection resisting medicine or feed. According to the application, research finds that dicoumarin has an obvious inhibition effect on eimeria for the first time, the effect can be observed on an in-vitro eimeria cell culture model, and an in-vivo infection model of an animal with chicken infected with eimeria is verified. The application of the dicoumarin to the field of parasitic infection resistance is beneficial to expanding the medicinal value of the dicoumarin.

Description

Technical Field

[0001] The present invention relates to the technical field of the application of dicoumarol, and specifically relates to the application of dicoumarol in the preparation of drugs or feeds for preventing and treating Eimeria infection. Background Art

[0002] Infection with Eimeria causes coccidiosis in chickens, which is the most common protozoal disease in chickens and causes great harm to the chicken farming industry. Coccidiosis has always been one of the main diseases to be controlled and treated in chicken production. Especially after the large-scale popularization and application of intensive chicken farming methods, it has become a common and frequently-occurring disease, among which Eimeria tenella is the most serious. Anticoccidial drugs can be said to be one of the drugs that must be added to feeds. However, when chemical drugs are used for a long time to prevent and treat chicken coccidiosis, the problem of drug resistance of coccidia is becoming increasingly serious. More than 20 drug-resistant strains of coccidia have been discovered globally, and the drug resistance rate in some areas is as high as over 80%. Problems such as coccidia drug resistance and drug residues have made the research of anticoccidial drugs enter a bottleneck stage, and there is an urgent need for new anticoccidial drugs to be put on the market.

[0003] Dicoumarol, also known as spoilt sweet clover factor, is an inhibitor of vitamin K epoxide reductase complex subunit 1. It mainly exists in spoiled sweet clover. During the spoilage process of alfalfa, its components react to form dicoumarol. Existing research shows that its main function is anticoagulation. By inhibiting the synthesis of vitamin K-dependent coagulation factors (coagulation factors II, VII, IX, X) in the liver, it interferes with the coagulation process, prevents the formation and expansion of thrombi, and is commonly used to treat and prevent thromboembolic diseases.

[0004] The molecular formula of dicoumarol is C 19 H 12 O6, with a molecular weight of 336.29 and a CAS number of: 66-76-2. Its chemical structure is shown as follows

[0005]

[0006] As a drug for preventing thrombus formation, preventing deep vein thrombosis, preventing pulmonary embolism, preventing complications of myocardial infarction, preventing complications of cerebral infarction, reducing the thrombus risk of atrial fibrillation, and preventing thrombus formation after cardiac valve replacement, dicoumarol has attracted much attention in recent years. However, there is currently no report on the anti-Eimeria effect of dicoumarol, and it is impossible to infer whether it has an anti-Eimeria effect based on the known properties of dicoumarol. Summary of the Invention

[0007] The technical problem to be solved by the present invention is how to propose a new use of dicoumarol.

[0008] The present invention solves the above technical problem by the following technical means:

[0009] The first aspect of the present invention provides the use of dicoumarol as an inhibitor of Eimeria, and the structural formula of the dicoumarol is as follows:

[0010]

[0011] The second aspect of the present invention provides the use of dicoumarol in the preparation of a drug or feed for preventing Eimeria infection.

[0012] Preferably, the feed includes complete formulated feed, concentrated feed, premix / premixed feed, concentrate mixture or mixed feed.

[0013] The third aspect of the present invention provides an anti-Eimeria product, the active ingredient of which includes dicoumarol.

[0014] Preferably, the product includes a drug or a feed additive.

[0015] Preferably, the product is a drug and further includes a pharmaceutically acceptable carrier.

[0016] "Pharmaceutically acceptable" means a non-toxic material that does not reduce the activity of the active ingredient. Such pharmaceutically acceptable buffers, carriers or excipients are well known in the art (see Remington’s Pharmaceutical Sciences, 18th Edition, edited by A.R. Gennaro, Mack Publishing Company (1990) and handbook of Pharmaceutical Excipients, 3rd Edition, edited by A. Kibbe, Pharmaceutical Press (2000)).

[0017] Preferably, the pharmaceutically acceptable carrier includes, but is not limited to, diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, disintegrants.

[0018] Preferably, the drug is made into a pharmaceutically acceptable dosage form.

[0019] Preferably, the dosage form includes tablets, capsules, granules, pills, dripping pills, syrups, powders, powders for external use, suppositories, drops, emulsions, solutions or suspensions.

[0020] Preferably, the administration method of the drug is group administration.

[0021] Preferably, the product is a feed additive, and its addition amount in the feed is 1-600 mg / kg.

[0022] Preferably, the addition amount is 10-200 mg / kg.

[0023] Preferably, the addition amount is 20 to 50 mg / kg.

[0024] The beneficial effects of the present invention are as follows:

[0025] (1) Through research, the present invention first discovers that dicoumarol has an obvious inhibitory effect on Eimeria, which can be observed in the cell culture model of Eimeria in vitro and is verified by the animal infection model of chickens infected with coccidia.

[0026] (2) At the feed addition dose of 20 to 50 mg / kg, the anticoccidial index (ACI) of dicoumarol can reach high efficiency (180), showing excellent anti-Eimeria effect and having no toxic effect on animals. In view of this discovery, the application field of dicoumarol is developed, providing a solution for the development of new anticoccidial drugs.

[0027] (3) Dicoumarol can effectively inhibit the development and proliferation of Eimeria tenella in the cell culture model of Eimeria tenella in vitro. Animal experiments further confirm that dicoumarol has a good anti-coccidial infection effect and can be used for the development of new anti-coccidial infection drugs or feeds.

[0028] (4) In the present invention, MDBK cells are cultured in DMEM medium containing 10% fetal bovine serum in vitro. When the cells grow to a confluence state of 80% or more, the extracted sporozoites of Eimeria tenella are inoculated into MDBK cells. After 4 h, the medium is discarded and washed 3 times with PBS to remove the sporozoites that have not invaded the cells. Then, DMEM medium containing 0.1 μg / ml, 0.5 μg / ml, 1 μg / ml, 3 μg / ml, 5 μg / ml, 10 μg / ml dicoumarol is added, and 3 replicates are set for each concentration. At the same time, a blank control group, a 0.1 μg / ml diclazuril control group, and an inoculated control group are set. It is placed in an incubator at 42 °C containing 5% CO2 and cultured continuously. After 48 h of culture, the medium is discarded and washed 3 times with PBS, and the total RNA of the samples in each well is extracted. The expression level of the Et actin gene of the surviving Eimeria tenella in each drug treatment group is detected by RT-qPCR to evaluate the effect of dicoumarol on coccidial proliferation in the in vitro coccidial culture model. The results show that the inhibition rate of the 0.1 μg / ml diclazuril treatment group on coccidia reaches 82.372%, proving that the culture model works well. The inhibition rates of each concentration group of dicoumarol on coccidia are between 17.73% and 77.36%, showing a good anti-Eimeria tenella effect.

[0029] (5) In the present invention, dicoumarol is made into a powder together with acceptable excipients, added to the feed, mixed evenly, and fed to chickens according to the daily feed amount. The addition amount of dicoumarol is 0.1 - 60 g of dicoumarol per 100 kg of feed (i.e., 1.0 - 600 mg / kg of feed), and at the same time, the feed containing 1 mg / kg of diclazuril and the drinking water containing 15 mg / L of 7-hydroxycoumarin are used as the control group. It was found that when the addition dose was 20 - 50 mg / kg, through the experimental model of chicken infected with coccidia in vivo, it was confirmed that the anti-coccidial index (ACI) of dicoumarol could reach high efficiency (180), having excellent effects against Eimeria tenella, and being safe and non-toxic to the fed animals.

[0030] Based on the above experimental results, both in vitro and in vivo studies have shown that dicoumarol can effectively inhibit the proliferation of coccidia and can be used for chickens to resist coccidial infection. Brief Description of the Drawings

[0031] Figure 1 It is a diagram showing the effect of dicoumarol on the proliferation of Eimeria in the in vitro culture model of Eimeria in Example 1 of the present invention; in the figure, the abscissa is the concentration of the compound dicoumarol, and the ordinate is the inhibition rate of the compound dicoumarol on Eimeria; in the figure, * represents P < 0.01, ** represents P < 0.005, *** represents P < 0.0005, and **** represents P < 0.0001.

[0032] Figure 2 It is a diagram showing the pathological effect of dicoumarol on the ceca of chickens infected with Eimeria after treatment in Example 2 of the present invention;

[0033] Figure 3 is Figure 2 the corresponding enlarged local view. Detailed Embodiments

[0034] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention. Unless otherwise defined, the professional terms used below have the same meaning as understood by those of ordinary skill in the art.

[0035] The test materials, reagents, etc. used in the following embodiments can be obtained from commercial channels or prepared by known methods without special instructions.

[0036] For those without specific technologies or conditions noted in the examples, they can all be carried out according to the technologies or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, for the quantitative tests in the following examples, more than three repeated experiments are set, and the results are averaged.

[0037] Example 1:

[0038] This example provides the anti - Eimeria effect of dicoumarol in an in vitro Eimeria infection model, specifically as follows:

[0039] I. Experimental materials:

[0040] (1) Drugs: Dicoumarol (≥98%) and diclazuril (≥98%) were purchased from Aladdin Biochemical Technology Co., Ltd.

[0041] (2) Oocysts: Sporulated oocysts of Eimeria tenella were preserved by the Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, and were rejuvenated in coccidia - free chicks before use.

[0042] (3) Cells: MDBK cells (i.e., bovine kidney cells) were preserved by the Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences.

[0043] II. Experimental methods:

[0044] In this example, the anti - Eimeria tenella effect of dicoumarol was evaluated using an Eimeria tenella - infected MDBK cell culture model. The specific evaluation steps include:

[0045] (1) Digest MDBK cells that have grown to a confluence state of over 80% with 0.25% trypsin digestion solution, plate them in a 12 - well cell culture plate at a cell concentration of 4×10 5 cells / well, and culture them in an incubator at 37°C with 5% CO2 for 24 h.

[0046] (2) When the cells in the 12 - well culture plate grow to a confluence state of 80% or more, inoculate 12×10 4 sporozoites / ml into each well. After 4 h, discard the culture medium, wash 3 times with PBS to wash away the suspended sporozoites, and add DMEM medium with 2% fetal bovine serum to each well.

[0047] (3) Experimental grouping:

[0048] The drug - to - be - detected group (adding dicoumarol) was set with 6 drug concentration gradients, specifically 0.1 μg / ml, 0.5 μg / ml, 1 μg / ml, 3 μg / ml, 5 μg / ml, and 10 μg / ml, and 3 replicates were set for each concentration;

[0049] The diclazuril treatment group (added diclazuril) was set with 4 drug concentration gradients, specifically 0.05 μg / ml, 0.1 μg / ml, 0.5 μg / ml, and 1 μg / ml, and 3 replicates were set for each concentration;

[0050] Blank control group: Referring to the volumes of the test drug and the solvent DMSO of diclazuril added in the test drug group and the diclazuril treatment group, an equal volume of DMSO and medium were added. This medium was DMEM medium with 2% fetal bovine serum, and 3 replicates were set.

[0051] (4) It was placed in an incubator at 42 °C with 5% CO2 and continued to be cultured. After 48 h of culture, the medium was discarded, washed 3 times with PBS, the total RNA of the samples in each well was extracted, and the expression of the actin gene of Eimeria tenella was detected by RT-qPCR. The effect of each test drug on the proliferation of Eimeria tenella was represented by the expression level of the actin gene.

[0052] Real-time qPCR primers are as follows:

[0053] Primers for specifically amplifying the actin of Eimeria tenella,

[0054] EtActin-F: 5’-CACCACCGCCGAGAAAGA-3’ (SEQ ID NO.1),

[0055] EtActin-R: 5’-GAACAACATTGCCGTAGAGG-3’ (SEQ ID NO.2);

[0056] Primers for specifically amplifying the actin of host cells,

[0057] Bactin-F: 5’-GGATGAGGCTCAGAGCAAGAGA-3’ (SEQ ID NO.3),

[0058] Bactin-R: 5’-TCGTCCCAGTTGGTGACGAT-3’ (SEQ ID NO.4).

[0059] Using the ΔΔC T method to calculate the effect of each test drug on the growth and development of Eimeria tenella at the cell culture level. The Real-time qPCR reaction system is shown in Table 1, and the Real-time qPCR reaction program is shown in Table 2.

[0060] Table 1 Real-time qPCR reaction system

[0061]

[0062]

[0063] Table 2 Real-time qPCR reaction program

[0064]

[0065] Evaluation index: Obtain ΔC of the experimental group and the control group by RT-qPCR T = C T [Et actin] - C T [βactin], ΔΔC T = ΔC T[exp] -ΔC T[ref] , and finally obtain the anti-Eimeria tenella effect of the drug to be screened:

[0066]

[0067] III. Experimental results:

[0068] Calculate the inhibitory effects of the drug to be detected and diclazuril treatment on Eimeria tenella respectively. Both coumarin and diclazuril treatment have obvious inhibitory effects on the reproduction of Eimeria tenella (Table 3). The results show that both coumarin and diclazuril have good anti-Eimeria tenella effects.

[0069] Table 3 Inhibitory effects of each test drug on coccidia reproduction at the cellular level

[0070]

[0071] The effect of coumarin on the proliferation and development of Eimeria tenella in the in vitro Eimeria tenella culture model is as Figure 1 shown. In the figure, the abscissa is the concentration of the compound coumarin, and the ordinate is the inhibition rate of the compound coumarin on the reproduction of Eimeria tenella. According to Figure 1 it can be seen that as the concentration of coumarin increases, the inhibitory effect of coumarin on the reproduction of Eimeria tenella increases. The effect of coumarin is concentration-dependent and has good activity against the development and reproduction of Eimeria tenella.

[0072] Example 2:

[0073] This example studied the safety range of coumarin in chickens, as follows:

[0074] I. Experimental materials:

[0075] (1) Chicks: 10-day-old Sanhuang broilers that are healthy and have similar weights.

[0076] (2) Preparation of coumarin-containing feed: Accurately weigh coumarin and add it to the blank feed step by step according to the designed dose, and prepare 10 kg of medicated feed for standby.

[0077] (3) Blank feed: The brooding feed was customized by Shanghai Wangyuan Feed Technology Co., Ltd. and did not contain any anticoccidial drugs.

[0078] (4) Coccidia oocysts: Sporulated oocysts of Eimeria tenella, preserved by the Shanghai Veterinary Research Institute, were rejuvenated in coccidia-free chicks before use.

[0079] II. Experimental methods:

[0080] (1) Grouping: 100 Sanhuang broilers were raised to 10 days old, weighed one by one, and weak chicks and those with too large body weight were eliminated. The remaining chicks were grouped using the random grouping method with excel software into 8 groups, with 10 chicks in each group, and the body weights of each group of chickens were adjusted to be approximately equal.

[0081] (2) Treatment: Chicks in groups 1 and 5 were fed normal feed every day; chicks in groups 2 - 4 and 6 - 8 were fed different doses of coumarin starting from 11 days old until 17 days old; chicks in groups 5 - 8 were orally administered 3×10 sporulated oocysts of Eimeria tenella per chick at 14 days old; the medicated feed was continuously fed for 7 days. The grouping of the experimental chickens and the dosage of the drug are shown in Table 4. 4 per chick; the medicated feed was continuously fed for 7 days. The grouping of the experimental chickens and the dosage of the drug are shown in Table 4.

[0082] Table 4 Experimental grouping

[0083]

[0084] (3) Result statistics:

[0085] Daily observation: Observe and record the mental state, feeding situation, drinking water situation, feather state, fecal characteristics, etc. of the chickens every day, as well as whether there are abnormal behaviors or deaths.

[0086] Body weight monitoring: Weigh the chickens in each group before and after feeding the medicated feed and record the weight changes.

[0087] Pathological examination: After the experiment, all the chickens were euthanized, and the pathological changes such as the appearance, size, color, and texture of the main organs such as the heart, liver, spleen, lungs, and kidneys were observed by dissection.

[0088] III. Experimental results:

[0089] Chicks in groups 1 - 4 that were not infected with Eimeria tenella were fed dicoumarol at various doses (200, 400, and 600 mg / kg of feed). None of the chicks died, but their feed intake and water consumption decreased, and there were signs of feather pecking and listlessness. Among the chicks in groups 5 - 8 that were infected with coccidia, those fed a high dose of dicoumarol (100 mg / kg of feed) had a large amount of bloody stools, feather pecking, and a mortality rate as high as 100%. Chicks fed medium and low doses (20 and 50 mg / kg of feed) did not die, but their feed intake decreased, and their weight gain rate was lower compared to the blank group. The specific results are shown in Table 5.

[0090] Table 5 Drug safety in chickens of each group

[0091]

[0092] In the toxicity test of dicoumarol in chickens, when 10 - day - old chicks were not infected with Eimeria tenella and fed drugs containing dicoumarol, no chicks died, but there were signs of feather pecking and bleeding. When the feed contained 400 mg / kg of dicoumarol, there were no such phenomena, indicating good safety. After the chicks were infected with coccidia at 14 days old and the feed contained 50 mg / kg of dicoumarol, no chicks died and the relative weight gain rate was normal, showing good safety. When dicoumarol is used as an anticoccidial drug, the dose should not exceed 50 mg / kg.

[0093] Example 3:

[0094] This example provides the effect and effective dose of dicoumarol against Eimeria tenella infection in chickens, as follows:

[0095] I. Experimental materials:

[0096] (1) Preparation of dicoumarol - containing feed: 2 g and 5 g of dicoumarol were respectively mixed with 100 kg of blank feed step by step, and finally feeds containing 20 mg / kg and 50 mg / kg of dicoumarol were obtained.

[0097] (2) Control drugs: The conventional anticoccidial drug diclazuril and 7 - hydroxycoumarin reported in the literature (patent for invention 201310471797.9) were used. Diclazuril was produced by Inner Mongolia Qihui Pharmaceutical Co., Ltd., and 7 - hydroxycoumarin was purchased from Aladdin Biochemical Technology Co., Ltd.

[0098] (3) Oocysts: Sporulated oocysts of Eimeria tenella were preserved by the Shanghai Veterinary Research Institute and were rejuvenated in coccidia - free chicks before use.

[0099] (4) Chicks: Sanhuang broiler chickens, provided by Shanghai Fuji Biotechnology Co., Ltd., were raised in a disinfected dedicated animal house; the chicken coops and utensils used were strictly disinfected, and they were allowed to feed freely and drink pure water; before the experiment, the chicks were observed for any clinical symptoms and the feces were examined for coccidia oocysts for 2 consecutive days, and then they were set aside for use.

[0100] (5) Blank feed: The starter feed was customized by Shanghai Wangyuan Feed Technology Co., Ltd. and did not contain any anticoccidial drugs.

[0101] II. Experimental methods:

[0102] (1) Grouping: 200 Sanhuang broiler chickens were raised to 10 days old according to the experimental grouping and treatment, weighed one by one, and the weak chicks and those with too large body weights were eliminated. The remaining chicks were grouped using the random grouping method with excel software into 6 groups, with 30 chicks in each group, and the body weights of the chicks in each group were appropriately adjusted to be approximately equal.

[0103] (2) Treatment: Except for the first group (blank control) that was fed the feed normally every day, the chicks in the second to fifth groups were orally administered 3×10 sporulated oocysts of Eimeria tenella per chick at one time; on the day of inoculation, the third to sixth groups were fed with anticoccidial drugs for 7 days respectively. The grouping of the experimental chickens and the usage and dosage of the drugs are shown in Table 6. 4 per chick; on the day of inoculation, the third to sixth groups were fed with anticoccidial drugs for 7 days respectively. The grouping of the experimental chickens and the usage and dosage of the drugs are shown in Table 6.

[0104] Table 6 Experimental grouping

[0105]

[0106] (3) Result statistics:

[0107] The mental state, feed intake, feces condition, etc. of the chicken flock were observed and recorded every day; the dead chicks were weighed and necropsied. If the death was caused by Eimeria tenella infection, the lesion score was recorded as +4 points; all the chicks were weighed one by one and necropsied on the 7th day after infection, the cecal lesions were scored, the ceca of each group were collected and fixed, and pathological sections were made by H.E. staining; finally, the weight gain and feed conversion rate of the chicks in each group were calculated.

[0108] Method and standard for judging drug efficacy The Anticoccidia Indexes (ACI) were calculated according to the formula recommended by Merck & Co., Inc. of the United States:

[0109] ACI = (relative weight gain rate + survival rate) - (oocyst value + lesion value);

[0110] Relative weight gain rate (%) = (weight gain of the experimental group ÷ weight gain of the blank control group) × 100%;

[0111] Survival rate (%) = (number of surviving chicks in each group ÷ total number of chicks in each group) × 100%.

[0112] The lesions were scored on a five - point scale:

[0113] ① Without oocysts and with normal ceca, scored 0 points;

[0114] ② With oocysts, slightly thickened cecal mucosa, with a small amount of scattered bleeding or a small amount of bloody intestinal contents, scored +1 point;

[0115] ③ With oocysts, thickened cecal mucosa, with obvious bleeding or obvious bloody intestinal contents, scored +2 points;

[0116] ④ With oocysts, thickened cecal mucosa, with a large number of blood clots or a bloody intestinal core, scored +3 points;

[0117] ⑤ Chicks died due to coccidiosis or had a large number of oocysts, the ceca appeared soy - sauce - colored (or there were punctate necrotic foci in the middle of the small intestine, and the mucosal surface was scarlet), the intestinal tube was significantly enlarged, and the contents formed an obvious bloody intestinal core, scored +4 points.

[0118] Lesion value = average lesion score of each group × 10.

[0119] The oocyst value was converted from the number of oocysts per gram of feces (OPG) in the cecal contents.

[0120] The criteria for judging the drug effect were: ACI < 120 was ineffective, 120 - 160 was low - effective, 160 - 180 was medium - effective, and above 180 was high - effective.

[0121] III. Experimental results:

[0122] (1) Observation of clinical symptoms: In the control group (Group 2) inoculated with sporulated oocysts of Eimeria tenella, on the 2nd day, the experimental chickens showed listlessness and a decrease in feed intake. On the 5th day, obvious bloody stools appeared and deaths occurred. In Groups 3 and 4, the appetite decreased slightly on the 3rd day after inoculation with sporulated oocysts of Eimeria tenella. In Group 1, the appetite and mental state were good during the experiment.

[0123] (2) The anti - coccidial indices of Groups 1 - 6 were calculated respectively, and the results are shown in Table 7. The anti - coccidial indices of Groups 5 and 6 (dicoumarol group) were higher than those of the control drug Group 4 (7 - hydroxycoumarin group), and the anti - coccidial index (ACI) of dicoumarol could reach 180, showing good anti - Eimeria tenella effect.

[0124] Table 7 Anti - coccidial indices of each group

[0125]

[0126] (4) After fixing the chicken ceca and making pathological sections, pathological changes such as mucosal integrity, degree of inflammation, and number of worms in the cecal tissues of each group were observed under an optical microscope as Figure 2 andFigure 3 As shown, the results indicate that: the dicoumarol group significantly alleviated cecal mucosal damage, reduced inflammatory cell infiltration, and significantly inhibited the development of coccidia, with a decrease in the number of worms. Although the 7-hydroxycoumarin group had a certain effect, the effect was not as significant as that of dicoumarol. Typical coccidia lesions, mucosal exfoliation, and a large number of oocysts were observed in the positive control group, while the dicoumarol group was close to the structure of the negative control group, confirming that dicoumarol can effectively relieve the cecal pathological damage caused by coccidia infection.

[0127] In summary, the embodiments of the present invention disclose that dicoumarol can be used to resist the infection of Eimeria tenella in chickens, expanding the uses of dicoumarol and developing new anti-coccidial drugs at the same time. The embodiments of the present invention have conducted cell-level tests and animal tests, and the results show that dicoumarol has a significant effect of inhibiting Eimeria tenella. The dicoumarol provided by the embodiments of the present invention can be used to prepare anti-Eimeria tenella drugs.

[0128] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of dicoumarol as an inhibitor of Eimeria, characterized in that, The structural formula of dicoumarol is as follows:

2. Use of dicoumarol in the preparation of a medicament or feed for preventing Eimeria infection.

3. An anti-Eimeria product, characterized in that, Its active ingredient includes dicoumarol.

4. The anti-Eimeria product according to claim 3, characterized in that, The product includes a medicament or a feed additive.

5. The anti-Eimeria product according to claim 4, wherein When the product is a medicament, it further includes a pharmaceutically acceptable carrier.

6. The anti-Eimeria product according to claim 4, wherein The pharmaceutically acceptable carrier includes a diluent, a binder, a surfactant, a wetting agent, an adsorbent carrier, a lubricant, a filler, and a disintegrant.

7. The anti-Eimeria product according to claim 4, characterized in that, The medicament is made into a pharmaceutically acceptable dosage form, and the dosage form includes tablets, capsules, granules, pills, dripping pills, syrups, powders, powders for external use, suppositories, drops, emulsions, solutions or suspensions.

8. The anti-Eimeria product according to claim 4, wherein When the product is a feed additive, its addition amount in the feed is 1-600 mg / kg.

9. The anti-Eimeria product according to claim 8, wherein The addition amount is 10-200 mg / kg.

10. The anti-Eimeria product according to claim 8, characterized in that, The addition amount is 20-50 mg / kg.

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