A kind of medicine composition for preventing and treating avian white crown disease

Abstract

The present invention provides a pharmaceutical composition for preventing and treating avian white crown disease, belonging to the technical field of chicken breeding. The pharmaceutical composition of the present invention is prepared by mixing 10 to 30 parts of Morin hydrate, 5 to 20 parts of artemisinic acid, 5 to 20 parts of quinic acid, and 5 to 20 parts of ginsenoside total saponin sulfate derivatives. It can effectively improve the phenomena of a sharp drop in egg production rate, an increase in soft-shell eggs, and an increase in mortality rate of sick laying hens in cages, so that the egg production rate, egg quality, and lean meat content can be restored to normal levels; enhance the disease resistance of poultry, reduce stress reactions caused by factors such as drastic changes in temperature and microbial invasion, and improve the immunity of animal bodies; the dosage is small and the curative effect is significant. The high-dose oral liquid of the pharmaceutical composition prepared by the present application was used to feed sick chickens, and the effective rate of treatment reached 100%, and the cure rate reached 94.00%.

Description

Technical Field

[0001] The invention relates to the technical field of chicken breeding, and in particular to a pharmaceutical composition for preventing and treating avian white crown disease. Background Art

[0002] Avian white crown disease (Leucocytozoonosis) is an intracellular parasitic protozoan disease caused mainly by Leucocytozoa carinii and Leucocytozoa sabie invading the tissue cells of blood and internal organs. Poultry and birds can be infected, and the disease is more common in summer and autumn. It is often endemic in southern my country, and even outbreaks may occur. Because the parasites invade the red blood cells of chickens and parasitize in them, temporary anemia is caused. Clinical manifestations are often pale combs, emaciation, watery white or green loose feces, decreased or even stopped egg production in adult chickens, stunted development of middle-aged chickens, and slow weight gain. Chicks aged 3 to 6 weeks have obvious symptoms and a high mortality rate. It is one of the main diseases that endanger the poultry industry.

[0003] Because chickens are in a stress-sensitive state under the intensive production model, their immune and intestinal functions are low, and their sensitivity to various pathogens is increased. Mycotoxin contamination in feed can also easily lead to an increase in the incidence and mortality rate of white crown disease in chickens. Therefore, repeated infection with white crown disease is prone to occur in caged chickens. In clinical practice, tadalafil, sulfonamides, etc. are commonly used in combination with other antibacterial and anti-inflammatory drugs to treat white crown disease in chickens. However, since sulfonamides have strong toxic and side effects on chicken kidneys, long-term repeated use will lead to serious drug resistance and drug residues, which can easily lead to reduced drug efficacy, prevention and control failures, and food safety issues. In addition, most traditional Chinese veterinary drugs commonly used in clinical practice are powders, which often have problems such as unreasonable components, excessively large particle size, poor palatability, slow release of active ingredients, low bioavailability, and slow effect, which cannot meet the development needs of modern intensive scale breeding industry.

[0004] Therefore, the development of new veterinary drug products that are highly effective, targeted, non-resistant, low-residue, easy to use, and low-priced is of great significance to the effective solution to chicken white crown disease. Summary of the invention

[0005] The object of the present invention is to provide a pharmaceutical composition for preventing and treating avian white crown disease.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a pharmaceutical composition for preventing and treating avian white crown disease.

[0008] Preferably, the pharmaceutical composition comprises the following raw materials in parts by weight: 10-30 parts of Morin hydrate, 5-20 parts of artemisinic acid, 5-20 parts of quinic acid, and 5-20 parts of sulfated derivatives of total ginsenosides.

[0009] Preferably, the preparation method of the sulfated derivatives of total ginsenosides is:

[0010] (1) precooling anhydrous pyridine and chlorosulfonic acid and mixing until a large amount of white or light yellow solid appears to obtain an esterification reagent;

[0011] (2) dissolving total ginsenosides in anhydrous formamide to obtain a ginsenoside solution;

[0012] (3) mixing the ginsenoside solution with the esterification reagent for reaction, and adjusting the pH to neutral after the reaction is completed;

[0013] (4) adding anhydrous ethanol to the product of step (3) to carry out static precipitation, taking the supernatant and adding anhydrous ether to separate the phases, and collecting the precipitated crystals.

[0014] (5) placing the crystals collected in step (4) into a dialysis bag for dialysis, recovering the non-permeated liquid in the dialysis bag, freezing and drying to obtain the sulfated derivatives of total ginsenosides.

[0015] Preferably, the volume ratio of chlorosulfonic acid to anhydrous pyridine in step (1) is 1:1.5-4.0.

[0016] Preferably, the mass volume ratio of the total ginsenosides and anhydrous formamide mixed in step (2) is 8 to 10:140.

[0017] Preferably, in step (3), the mixing volume ratio of the esterification reagent to the ginsenoside solution is 1:1.5-3.0, the temperature of the mixing reaction is 40-50°C, and the time is 2-4h.

[0018] Preferably, the standing time in step (4) is 2 to 3 hours.

[0019] Preferably, the molecular weight cut-off of the dialysis bag in step (5) is 500 to 1000 Da.

[0020] Preferably, the pharmaceutical composition can be prepared as a solution, a water-soluble powder or a granule.

[0021] The pharmaceutical composition for preventing and treating avian white crown disease provided by the present invention is based on the causes and clinical characteristics of chicken white crown disease, with insecticidal blood-tonifying, anti-inflammatory and antibacterial, and immunomodulatory treatment principles, and effective monomers of traditional Chinese medicine are selected for combined use. It is prepared by mixing a certain weight portion of mulberry yellow hydrate, artemisinic acid, quinic acid, and sulfated ginseng total saponin derivatives. The pharmaceutical composition has an ideal insecticidal and antidiarrheal effect, a short treatment time, can reduce the use of sulfonamide drugs and antibiotics, and has no drug withdrawal period. It can effectively improve the phenomena of a sharp drop in the egg production rate of caged sick laying hens, an increase in soft-shell eggs, and an increase in the mortality rate, so that the egg production rate, egg quality, and lean meat content return to normal levels, enhance the disease resistance of poultry, reduce the stress response caused by factors such as drastic changes in temperature and microbial invasion, and improve the immunity of animal bodies. It has the characteristics of small dosage and significant therapeutic effect.

[0022] The effective rate of treating white crown disease in chickens by feeding the pharmaceutical composition prepared by the present application reached 89.67%, and the cure rate reached 84.67%. Compared with tadalafil, the effective rate increased by 11.10%, and the cure rate increased by 19.25%. The effective rate of treating white crown disease in chickens by feeding the high-dose pharmaceutical composition oral liquid prepared by the present application reached 100%, and the cure rate reached 94.00%. Compared with tadalafil, the effective rate increased by 8.5%, and the cure rate increased by 11%. DETAILED DESCRIPTION

[0023] The invention provides a pharmaceutical composition for preventing and treating avian white crown disease. The pharmaceutical composition comprises the following raw materials in parts by weight: 10 to 30 parts of mulberry yellow hydrate, 5 to 20 parts of artemisinic acid, 5 to 20 parts of quinic acid, and 5 to 20 parts of sulfated ginsenoside derivatives.

[0024] In the present invention, the pharmaceutical composition preferably comprises the following raw materials in parts by weight: 20 parts of Morin hydrate, 15 parts of artemisinic acid, 15 parts of quinic acid, and 10 parts of sulfated total ginsenoside derivatives.

[0025] In the present invention, the mulberry yellow indole hydrate (Lot#M813301, analytical grade AR) was purchased from Shanghai McLean Biochemical Technology Co., Ltd.; the artemisinic acid (Lot#C15H2202, ≧98%) was purchased from Guangdong Wengjiang Chemical Reagent Co., Ltd.; the quinic acid (Lot#D817144, C10532503, ≧98%) was purchased from Shanghai McLean Biochemical Technology Co., Ltd.

[0026] In the present invention, the preparation method of the sulfated derivatives of total ginsenosides is as follows:

[0027] (1) precooling anhydrous pyridine and chlorosulfonic acid and mixing until a large amount of white or light yellow solid appears to obtain an esterification reagent;

[0028] (2) dissolving total ginsenosides in anhydrous formamide to obtain a ginsenoside solution;

[0029] (3) mixing the ginsenoside solution with the esterification reagent for reaction, and adjusting the pH to neutral after the reaction is completed;

[0030] (4) adding anhydrous ethanol to the product of step (3) to carry out static precipitation, taking the supernatant and adding anhydrous ether to separate the phases, and collecting the precipitated crystals;

[0031] (5) placing the crystals collected in step (4) into a dialysis bag for dialysis, recovering the non-permeated liquid in the dialysis bag, freezing and drying to obtain the sulfated derivatives of total ginsenosides.

[0032] In the present invention, the precooling temperature of the anhydrous pyridine is preferably 0°C to 4°C, more preferably 0°C.

[0033] In the present invention, the mixing of anhydrous pyridine and chlorosulfonic acid is preferably carried out in a three-necked flask equipped with a stirring device and a condensing device.

[0034] In the present invention, the three-necked flask is preferably placed in an ice-salt water bath.

[0035] In the present invention, the temperature of the ice-salt bath is preferably -10°C to 0°C, more preferably -10°C.

[0036] In the present invention, the temperature at which the anhydrous pyridine and chlorosulfonic acid are mixed is -10°C to 0°C, preferably -10°C.

[0037] In the present invention, the volume ratio of chlorosulfonic acid to anhydrous pyridine is 1:1.5-4.0, preferably 1:2.5.

[0038] In the present invention, chlorosulfonic acid is added dropwise into anhydrous pyridine under stirring, and the stirring speed is 200-400 rpm, preferably 300 rpm.

[0039] In the present invention, the chlorosulfonic acid is added dropwise within 30 to 40 minutes, preferably within 40 minutes, until a large amount of white or light yellow solid appears, and the esterification reagent is prepared. The esterification reagent is generally prepared on the spot, and the esterification reagent can be sealed and stored at -20°C for short-term use.

[0040] In the present invention, the mass volume ratio of the total ginsenosides and anhydrous formamide is 8 to 10:140, preferably 10:140.

[0041] In the present invention, the ginsenosides are mixed with anhydrous formamide and stirred at 20° C. for 25 to 35 minutes, preferably 30 minutes, to obtain a ginsenoside solution.

[0042] In the present invention, the total ginsenosides are purchased from Jilin Hongjiu Biotechnology Company, with a purity greater than 90%.

[0043] In the present invention, the volume ratio of the esterification reagent to the ginsenoside solution is 1:1.5-3.0, preferably 1:2.

[0044] In the present invention, the temperature of the mixed reaction of the esterification reagent and the ginsenoside solution is 40-50°C, preferably 45°C; the time is 2-4h, preferably 2h. The mixed reaction needs to be stirred continuously, and the stirring speed is preferably 200-400rpm, more preferably 300rpm.

[0045] In the present invention, after the reaction is completed, the reaction product is preferably cooled to 20° C. and adjusted to neutrality with 2 mol / L NaOH in an ice bath.

[0046] In the present invention, the amount of anhydrous ethanol added is 2 to 4 times the volume of the reaction product of step (3), preferably 3 times the volume.

[0047] In the present invention, the standing time is 2 to 3 hours, preferably 2.5 hours.

[0048] In the present invention, after standing, the supernatant is preferably obtained by centrifugal separation, the rotation speed of the centrifuge is preferably 2500-3500 rpm, more preferably 3000 rpm, and the time of the centrifugation is preferably 8-12 min, more preferably 10 min.

[0049] In the present invention, the standing time after adding anhydrous ether for phase separation is preferably 2 to 3 hours, more preferably 2.5 hours.

[0050] In the present invention, the crystals precipitated after standing are preferably separated by centrifugation, the rotation speed of the centrifuge is preferably 2500-3500 rpm, more preferably 3000 rpm, and the time of the centrifugation is preferably 8-12 min, more preferably 10 min.

[0051] In the present invention, the process of static precipitation of anhydrous ethanol and phase separation of anhydrous ether is preferably repeated 2 to 4 times, more preferably 3 times, until no crystals are precipitated in the aqueous phase.

[0052] In the present invention, the molecular weight cut-off of the dialysis bag is 500-1000 Da, preferably 500 Da.

[0053] In the present invention, the dialysis is preferably performed with tap water for 3 days and then with deionized water for 1 day.

[0054] In the present invention, the pharmaceutical composition can be prepared as a solution, a water-soluble powder, or a granule.

[0055] In the present invention, the solution is a drug dissolved in water. Preferably, it includes an oral dosage form. This dosage form is mixed more evenly and can be put into drinking water to dissolve the drug, disperse it evenly, and provide it for animals to drink. This dosage form is aimed at the characteristics of sick chickens that have reduced appetite (even loss of appetite), increased body temperature and like to drink water, and can ensure that sick chickens can obtain enough drugs.

[0056] In the present invention, the pharmaceutical composition is in powder form, has good water solubility, and is stable in nature. It can be put into drinking water as a water-soluble powder to dissolve the drug, evenly disperse it, and provide it for animals to drink. This dosage form has a good effect on the prevention and treatment of diseases in chicken groups. In the present invention, the powder is a drug dosage form in which the drug is ground into powder and can be mixed in feed or dissolved in water and fed to animals. This dosage form has relatively stable quality and is widely used. It is mostly used in feed as an additive, as a clinical preventive and therapeutic drug, and can also be directly used in the feed production process.

[0057] In the present invention, the pharmaceutical composition is preferably further added with auxiliary materials.

[0058] In the present invention, when the pharmaceutical composition is made into an oral liquid, each 100 mL is equivalent to 1 g of the original mixed crude drug; after the various component drugs are evenly mixed, they become powdered drugs, which can be used as powder mixing materials, and each 1 g is equivalent to 1 g of the original mixed crude drug.

[0059] In the present invention, the pharmaceutical preparation is used in the prevention and treatment of white crown disease in chickens, and can be used in drinking water or mixed with feed. The added dosage is 1 part of oral liquid preparation 100mL / L drinking water (v / v), or 2g powder / 5kg chicken feed (w / w), and 3 to 5 days is a course of treatment.

[0060] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0061] Example 1

[0062] Preparation method steps of ginsenosides by sulfate modification

[0063] (1) Anhydrous pyridine precooled at 0°C is placed in a three-necked flask equipped with a stirring device and a condensing device, and placed in an ice-salt bath at -10°C. Chlorosulfonic acid is added dropwise to the anhydrous pyridine over 40 minutes, while the reactants are rapidly stirred at 300 rpm until a large amount of white or light yellow solids appear. After the dropwise addition reaction is completed, the ice bath is removed and the mixture is allowed to stand for 30 minutes to obtain an esterification reagent.

[0064] (2) Accurately weigh 10 g of ginsenosides (Jilin Hongjiu Biotechnology Co., Ltd., purity greater than 90%), dissolve it in 140 mL of anhydrous formamide at 20° C. using a magnetic stirrer, stir thoroughly for 30 min, and mix to obtain a ginsenoside solution.

[0065] (3) Pour the ginsenoside solution into the flask containing the esterification reagent, stir in a constant temperature water bath at 45°C for 2 h, and rotate at 300 rpm. After the reaction is completed, take out the flask, cool the reaction product to 20°C, and adjust the pH to neutral with 2 mol / L NaOH in an ice bath at 0°C.

[0066] (4) Add 3 times the volume of anhydrous ethanol to the product of step (3) for precipitation, let it stand for 2.5 hours, then centrifuge at 3000 rpm for 10 minutes, take the supernatant and add anhydrous ether equal to the supernatant for phase separation, let it stand for 2.5 hours, then centrifuge at 3000 rpm for 10 minutes, repeat the precipitation and phase separation process 3 times until no crystals are precipitated in the aqueous phase. Collect the separated precipitate and put it into a 500Da dialysis bag, dialyze it with tap water for 3 days, and dialyze it with deionized water for 1 day. Recover the non-permeated liquid in the dialysis bag, freeze and dry it to obtain the sulfate modified derivative.

[0067] According to the above preparation method, this example designed 9 treatment schemes based on the volume ratio of anhydrous pyridine and chlorosulfonic acid (1:1.5, 1:2.5, 1:4.0), the reaction temperature of ginseng total saponins and esterification reagent (45, 60, 85°C), and the reaction time (2h, 3h, 4h) using a three-factor three-level orthogonal experiment, obtained 9 derivatives, and named the 9 different conditions in the order of 1 to 9.

[0068] After the experiment, the yield, sulfate content and sulfate substitution degree of each group of sulfate-modified derivatives were measured, and the results are shown in Table 1. From Table 1, it can be seen that the yield, sulfate content and sulfate substitution degree of sulfate-modified ginsenoside derivative 6 are the best. It is used in the following examples.

[0069] Table 1 Effects of reaction temperature and time, chlorosulfonic acid-pyridine ratio on sulfation of total ginsenosides

[0070]

[0071]

[0072] Note: Yield (%) = product mass / total saponin mass × 100%.

[0073] Example 2

[0074] Each portion of the pharmaceutical composition for treating chicken white crown disease includes: 1g of Morin hydrate, 2g of artemisinic acid, 1g of quinic acid, and 2g of sulfated derivatives of total ginsenosides of ginseng.

[0075] After the drugs are mixed evenly, they are packaged in heat-sealed small aluminum foil bags (6.0 g / bag), labeled to obtain a drug composition powder, and stored at 4° C. for future use.

[0076] Oral solution: 1g of powder, 10mL of anhydrous ethanol (solubilizer and stabilizer), 2mL of benzyl alcohol (antibacterial agent), and sterile water to make up to 100mL. (Each 100mL of oral solution is equivalent to 1g of the original mixed herbal medicine).

[0077] Example 3

[0078] Each portion of the pharmaceutical composition for treating chicken white crown disease includes: 3g of Morin hydrate, 0.5g of artemisinic acid, 2g of quinic acid, and 0.5g of sulfated derivatives of total ginsenosides of ginseng.

[0079] After the drugs are mixed evenly, they are packaged in heat-sealed small aluminum foil bags (6.0 g / bag), labeled to obtain a drug composition powder, and stored at 4° C. for future use.

[0080] Oral solution: 1g of powder, 10mL of anhydrous ethanol (solubilizer and stabilizer), 2mL of benzyl alcohol (antibacterial agent), and sterile water to make up to 100mL. (Each 100mL of oral solution is equivalent to 1g of the original mixed herbal medicine).

[0081] Example 4

[0082] Each portion of the pharmaceutical composition for treating chicken white crown disease includes: 1.5g of mulberry yellow indole hydrate, 2g of artemisinic acid, 0.5g of quinic acid, and 2g of sulfated derivatives of total ginsenosides of ginseng.

[0083] After the drugs are mixed evenly, they are packaged in heat-sealed small aluminum foil bags (6.0 g / bag), labeled to obtain a drug composition powder, and stored at 4° C. for future use.

[0084] Oral solution: 1g of powder, 10mL of anhydrous ethanol (solubilizer and stabilizer), 2mL of benzyl alcohol (antibacterial agent), and sterile water to make up to 100mL. (Each 100mL of oral solution is equivalent to 1g of the original mixed herbal medicine).

[0085] Example 5

[0086] Each portion of the pharmaceutical composition for treating chicken white crown disease includes: 2g of Morin hydrate, 1.5g of artemisinic acid, 1g of quinic acid, and 1.5g of sulfated derivatives of total ginsenosides of ginseng.

[0087] After the drugs are mixed evenly, they are packaged in heat-sealed small aluminum foil bags (6.0 g / bag), labeled to obtain a drug composition powder, and stored at 4° C. for future use.

[0088] Oral solution: 1g of powder, 10mL of anhydrous ethanol (solubilizer and stabilizer), 2mL of benzyl alcohol (antibacterial agent), and sterile water to make up to 100mL. (Each 100mL of oral solution is equivalent to 1g of the original mixed herbal medicine).

[0089] Example 6

[0090] Each portion of the pharmaceutical composition for treating chicken white crown disease includes: 2g of Morin hydrate, 1.5g of artemisinic acid, 1.5g of quinic acid, and 1g of sulfated derivatives of total ginsenosides of ginseng.

[0091] After the drugs are mixed evenly, they are packaged in heat-sealed small aluminum foil bags (6.0 g / bag), labeled to obtain a drug composition powder, and stored at 4° C. for future use.

[0092] Oral solution: 1g of powder, 10mL of anhydrous ethanol (solubilizer and stabilizer), 2mL of benzyl alcohol (antibacterial agent), and sterile water to make up to 100mL. (Each 100mL of oral solution is equivalent to 1g of the original mixed herbal medicine).

[0093] Example 7

[0094] Clinical application of pharmaceutical compositions

[0095] A laying hen farm in Huadu, Guangzhou, Guangdong Province, has a total of 6 chicken houses, each with 10,000 Hailan brown laying hens, 200 days old. Starting in June 2021, sick chickens showed depression, fever, loss of appetite, drooling and shaking heads, and excreted yellow-green loose feces. Most sick chickens showed anemia, pale combs and wattles, difficulty breathing, and mild paralysis of limbs and legs in the later stage. Postmortem features: emaciated corpse, pale muscles, white combs, thin blood, and poor blood coagulation. Extensive bleeding in internal organs, the liver, kidneys, and lungs are the most serious. The liver and spleen are hemorrhagic, enlarged, brittle and fragile, with small yellow-white nodules and hemorrhages on the surface. There are a large number of blood bags in the uterus, fallopian tubes, intestinal serosa, and pancreas of laying hens, which are suspected to be the direct cause of the decline in egg production in the farm.

[0096] Etiological diagnosis was performed on the patient, blood smears or organ slides were prepared, and Wright or Gibbs stained the patient. Microscopic examination revealed schizonts and round gametocytes of varying sizes in the white blood cells, as well as purple dot-shaped schizonts free in the plasma. Bacteriological isolation and culture did not show any bacterial growth. The case was diagnosed as chicken white crown disease after comprehensive diagnosis.

[0097] 1000 symptomatic sick chickens were randomly selected, grouped and administered according to Table 2, and treated with the oral liquid or powder prepared in Example 6 of the present invention. After the experiment, the mortality, inefficiency, effectiveness and cure rate of each group were calculated. Positive control therapeutic drug compound tadalafil soluble powder (product batch number: 20210426; specification: 100g / bag, purchased from Guangzhou Anxiu Animal Pharmaceutical Co., Ltd.), usage and dosage: 0.5g / kg mixed with feed, used for 5 to 7 days.

[0098] During the treatment period, the clinical symptoms, comb color, appetite, drinking water and weight gain recovery of the chickens in each group were observed daily. The effective rate, mortality rate and cure rate of each group were recorded. The dead chickens were autopsied, and the liver or blood samples were collected for staining and microscopic examination for parasite etiology diagnosis. The observation was carried out until the 7th day after drug withdrawal, and the results are shown in Table 2.

[0099] Judging criteria:

[0100] (1) Mortality rate: After drug administration, clinical death occurs, and the percentage of deaths to the total number of test chickens in the group is the mortality rate.

[0101] (2) Inefficiency: After medication, clinical symptoms do not improve, and even worsen or cause death, which is ineffective. The percentage of ineffective chickens to the total number of test chickens in the group is ineffective.

[0102] (3) Effectiveness: After medication, clinical symptoms are alleviated but not completely eliminated, and physiological indicators such as feeding, drinking, and spirit tend to be normal. The effective rate is the percentage of the effective number to the total number of test chickens in the group.

[0103] (4) Cure rate: After medication, clinical symptoms completely disappeared and there was no recurrence within 7 days of stopping medication. The cure rate was the percentage of the number of cured chickens to the total number of test chickens in the group.

[0104] Table 2 Observation test of drug efficacy of chicken white crown disease

[0105]

[0106] As shown in Table 2, after adding the pharmaceutical composition of the present invention to the drinking water or feed of the diseased laying hens and continuously administering the drug for 5 days, the treatment was carried out according to the experimental group 2 to 4 schemes. The cure rate of the oral liquid and powder pharmaceutical compositions for the diseased laying hens was significantly higher than that of the tamidin control group, and the treatment effect was relatively ideal. The effective results of the drug treatment groups 2 and 3 were similar. However, the mortality rate of group 3 was significantly better than that of group 2. Secondly, 66% of the laying hens in the infected control group were still in the diseased state, and the mortality rate reached 34%. 7 days after the end of the drug administration, the egg production rate of the laying hens in each drug administration group basically returned to the level before the onset of the disease, and the recovery of the oral liquid preparation was better than that of the powder.

[0107] After 3 days of administration, the mental state and appetite of the sick chickens in the three drug treatment groups improved significantly, the antidiarrheal effect was obvious, and no yellow-green loose feces were discharged. After 2 days of continued administration, the clinical symptoms and eating conditions of the sick chickens basically returned to normal. In order to improve the accuracy of the test, prevent repeated infections, and cut off the transmission of the disease, pyrethroids were used to spray and disinfect the chicken house (0.02%, 1 time / d, for 1 week) to kill the disease vectors such as Culicoides and blackflies inside and outside the chicken house. At the same time, the chicken manure was piled up and fermented to be harmless, and gauze was nailed on the doors and windows of the chicken house to reduce insect invasion. In the follow-up, the chickens were healthy and there was no recurrence of chicken white crown disease.

[0108] Example 8

[0109] In August 2021, a broiler farm in Liaocheng City, Shandong Province, had a total of 2 chicken houses, each with 5,000 white-feathered broilers, 30 days old. They were kept in pollution-free cages at room temperature of 20-24°C and relative humidity of 50±10%; the feed was complete feed for broiler chicks (drug-free compound feed), fed 3 times a day, and had free access to drinking water. Among them, 866 broilers became ill (39 died), and they were diagnosed with chicken white crown disease based on comprehensive clinical symptoms, autopsy observations, and organ smear staining and microscopy (the same etiological diagnosis as in Example 6). With the consent of the farmers, all sick chickens were used for this treatment trial, and the farmers assisted the experimenters in randomly grouping the sick chickens and conducting efficacy trials against chicken white crown disease.

[0110] In order to verify the clinical therapeutic effect of the present invention on chicken white crown disease, the pharmaceutical composition (oral liquid preparation) of Example 6 of the present invention was divided into three doses of high, medium and low and administered to broilers in groups to evaluate the clinical efficacy and optimal dosage of the present invention. The results are shown in Table 3.

[0111] Drug treatment groups and doses:

[0112] A-high-dose treatment group (200 mL / L): 2 portions of drug composition (oral solution) / 1 L of drinking water (w / v), 5 days as one course of treatment.

[0113] B-medium-dose treatment group (100 mL / L): 1 portion of drug composition (oral solution) / 1 L of drinking water (w / v), 5 days as one course of treatment.

[0114] C-low-dose treatment group (50 mL / L): 0.5 portions of the drug composition (oral solution) / 1 L of drinking water (w / v), 5 days as one course of treatment.

[0115] D-control drug treatment group: Compound tadalafil soluble powder (product batch number: 20210426; specification: 100g / bag, purchased from Guangzhou Anxiu Animal Pharmaceutical Co., Ltd.) Usage and dosage: 2g / L drinking water (w / v), continuous use for 5-7 days.

[0116] E-infection control group: no medication, normal feeding and drinking water.

[0117] F-Healthy control group: No medication, normal feeding and drinking water, and separate isolation.

[0118] Efficacy determination and observation indicators:

[0119] During the treatment period, the comb color, clinical symptoms, appetite, drinking water and weight gain recovery of the chickens in each group were observed daily. The effective rate, mortality rate and cure rate of each group were recorded. The dead chickens were dissected, and liver or blood samples were collected for staining and microscopic examination for parasite etiology diagnosis. The observation lasted until the 7th day after drug withdrawal.

[0120] Death: During the test period, chickens showed typical symptoms of white crown disease and died. The autopsy showed typical pathological characteristics. Microscopic examination of the main visceral target organ smears and blood smears showed schizonts and round gametocytes of varying sizes in the white blood cells. Free purple dot-shaped schizonts were seen in the plasma. It can be determined that the chickens died due to infection with the parasite. The mortality rate of each group was calculated based on the number of dead chickens.

[0121] Ineffective: There was no significant improvement in the comb color, breathing, spirit, and appetite of sick chickens.

[0122] Effective: After taking the medicine, the clinical symptoms of the chickens improved significantly, the diarrhea symptoms were alleviated, and the comb color, mental state, appetite, breathing, etc. gradually returned to normal, which was judged to be effective.

[0123] Cure: After taking the medicine, the chickens' comb color, mental state, and appetite returned to normal, yellow-green loose stools no longer occurred, and typical symptoms such as villus contamination around the anus and inflammation around the anus disappeared. There was no recurrence within 7 days of stopping the medicine, and the chickens were judged to be cured.

[0124] Relative weight gain rate: Calculate the average weight gain based on the weight of each chicken at the beginning and end of the experiment, and then calculate the average weight gain of each group of chickens. The weight gain of the blank control group is taken as 100%, and the weight gain of each group is compared with it to obtain the relative weight gain rate of each group.

[0125] Table 3 The efficacy and production performance results of each drug treatment group on chicken white crown disease (Mean±SD)

[0126]

[0127] Note: Compared with the infected control group, *, P<0.05; **, P<0.01

[0128] After 3 days of medication, compared with the infected control group, each drug treatment can quickly alleviate diarrhea symptoms, and the effect is fast. No dead chickens were found in the high and medium dose groups, and the mental state and appetite basically returned to normal. After 5 days of administration, the appetite returned to normal, and yellow-green loose stools were no longer discharged. The weight gain effect of the chickens in the drug treatment group was obvious, indicating that the pharmaceutical composition provided by the present invention has a good therapeutic effect on chicken white crown disease, small toxic and side effects, and significant weight gain effect. And the cure rate of the high and medium dose treatment groups of the pharmaceutical composition was significantly higher than that of the control drug tadalafil group, and the cure rate of the low dose group was slightly lower than that of the tadalafil group. The mental state and appetite recovery of the non-dead chickens in the infected control group were poor, the course of the disease was long, and the growth was slow. The weight gain effect of the high, medium and low treatment groups was not significantly different from that of the tadalafil group, and the feed-to-weight ratio results of each drug treatment group were significantly lower than those of the infected control group, among which the feed-to-weight ratio results of the medium dose drug treatment group and the healthy control group were close.

[0129] The pharmaceutical composition prepared by the present application was used to feed sick chickens, and the effective rate of treating white crown disease reached 89.67%, and the cure rate reached 84.67%. Compared with tadalafil, the effective rate increased by 11.1%, and the cure rate increased by 19.25%. After feeding the oral solution prepared by the pharmaceutical composition at a dose of 100mL / L to 200mL / L, the mental state and appetite of the sick chickens improved significantly after 3 days of administration, the antidiarrhea effect was obvious, and no yellow-green loose feces were discharged. After 2 days of continued administration, the clinical symptoms and dietary conditions of the sick chickens basically returned to normal. 7 days after the end of the administration, the egg production rate and feed-to-weight ratio of laying hens basically returned to the pre-onset level, the effective rate of treatment reached 100%, and the cure rate reached 94.00%.

[0130] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for preventing and treating avian white crown disease, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 10-30 parts of Morin hydrate, 5-20 parts of artemisinic acid, 5-20 parts of quinic acid, and 5-20 parts of sulfated ginsenoside derivatives; The preparation method of the sulfated derivatives of ginsenosides is as follows: (1) Precooled anhydrous pyridine and chlorosulfonic acid are mixed until a large amount of white or light yellow solid appears to obtain an esterification reagent; (2) dissolving total ginsenosides in anhydrous formamide to obtain a ginsenoside solution; (3) mixing the ginsenoside solution with the esterification reagent for reaction, and after the reaction is completed, adjusting the pH to neutral; (4) adding anhydrous ethanol to the product of step (3) to allow static precipitation, taking the supernatant and adding anhydrous ether to separate the phases, and collecting the precipitated crystals; (5) placing the crystals collected in step (4) into a dialysis bag for dialysis, recovering the non-permeated liquid in the dialysis bag, freezing and drying to obtain the sulfated derivatives of total ginsenosides; the dialysis bag has a molecular weight cutoff of 500-1000 Da.

2. The pharmaceutical composition according to claim 1, characterized in that The volume ratio of chlorosulfonic acid to anhydrous pyridine in step (1) is 1:1.5-4.

0.

3. The pharmaceutical composition according to claim 1, characterized in that The mass volume ratio of the total ginsenosides and anhydrous formamide mixed in step (2) is 8-10:

140.

4. The pharmaceutical composition according to claim 1, characterized in that In step (3), the mixing volume ratio of the esterification reagent to the ginseng total saponin solution is 1:1.5-3.0, the temperature of the mixing reaction is 40°C-50°C, and the time is 2-4h.

5. The pharmaceutical composition according to claim 1, characterized in that: The standing time in step (4) is 2 to 3 hours.

6. The pharmaceutical composition according to any one of claims 1 to 5, characterized in that The pharmaceutical composition is prepared in the form of solution, water-soluble powder or granules.

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