Traditional Chinese veterinary medicine oral liquid for preventing and treating damp-heat diarrhea of chicken as well as preparation method and application of traditional Chinese veterinary medicine oral liquid

Abstract

The invention belongs to the technical field of traditional Chinese veterinary medicines, and particularly relates to traditional Chinese veterinary medicine oral liquid for preventing and treating chicken diarrhea due to damp-heat as well as a preparation method and application. Active components of the traditional Chinese veterinary medicine oral liquid for preventing and treating chicken damp-heat diarrhea are prepared from the following raw materials in parts by weight: 4.5-9 parts of sweet wormwood herb, 9-18 parts of light bamboo shavings, 4.5-9 parts of rhizoma pinelliae preparata, 9-18 parts of red poria cocos, 4.5-9 parts of radix scutellariae, 4.5-9 parts of fructus aurantii, 4.5-9 parts of pericarpium citri reticulatae, 6-12 parts of talc, 2-4 parts of honeysuckle flower, 6-12 parts of herba portulacae and 1-2 parts of liquorice root. The traditional Chinese veterinary medicine oral liquid is simple to prepare, is specially used for preventing and treating damp-heat diarrhea of chickens, has a remarkable curative effect, can be taken by drinking water, is convenient to take for poultry, is high in absorption and utilization degree and free of medicine residues, and has important significance on guaranteeing green and healthy poultry breeding.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese veterinary medicines, and particularly relates to a traditional Chinese veterinary medicine oral liquid for preventing and treating damp-heat diarrhea in chickens, and a preparation method and application thereof. Background Art

[0002] Damp-heat diarrhea is a type of diarrhea. It refers to the diarrhea caused by damp-heat evil, which damages the stomach and intestines, causing abnormal digestion and digestion, and water and dampness flowing into the large intestine. The specific symptoms are diarrhea and abdominal pain, urgent diarrhea, or diarrhea without discomfort, yellow and foul-smelling feces, and burning in the anus. Damp-heat diarrhea in poultry is one of the most common diarrheal diseases. It affects the production performance of poultry, as well as the health and environmental hygiene of poultry, resulting in low feed conversion rate and decreased survival rate, becoming an important factor restricting the production efficiency of the poultry industry.

[0003] In the prevention and control of damp-heat diarrhea, farmers currently generally use Western medicines such as antibiotics for treatment. However, the long-term use of antibiotics and other chemical drugs has also brought about problems such as difficult to control veterinary drug residues and bacterial resistance, which in turn harms food safety and human health. The problems of drug residues and drug resistance caused by antibiotics and other chemical drugs directly restrict the sustainable development of the breeding industry and bring severe challenges to disease prevention and control. At present, the measures of reducing, limiting and banning antibiotics are increasing year by year. Therefore, there is a huge market demand for the research and development of alternative veterinary drug products. Traditional Chinese medicine has not only accumulated rich clinical experience in the treatment of diarrheal diseases, but also formed a unique method of syndrome differentiation and treatment, which has significant clinical efficacy. At the same time, Chinese veterinary medicine has won wide recognition and popularity in the Chinese and foreign veterinary medicine and animal husbandry circles in recent years for its natural source of medicine, integrity of efficacy, low toxicity and side effects, low residue, and not easy to cause pathogenic microorganisms to develop resistance. Research and development of safe and efficient Chinese veterinary medicine products for the prevention and treatment of diarrhea is of great significance to ensuring my country's green and healthy breeding. Summary of the invention

[0004] In order to solve the above problems, a Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens, a preparation method and application are provided. The liquid has simple preparation, reasonable proportions of the ingredients, good efficacy, no drug residues, easy absorption by poultry and convenient use, thus solving the problems existing in the prior art.

[0005] The invention provides a Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens. The active ingredients of the oral liquid are prepared from the following raw materials in parts by weight: 4.5-9 parts of Artemisia annua, 9-18 parts of Rhizoma Anemarrhenae, 4.5-9 parts of Rhizoma Pinelliae, 9-18 parts of Rhizoma Poriae, 4.5-9 parts of Radix Scutellariae, 4.5-9 parts of Fructus Aurantii Immaturus, 4.5-9 parts of Pericarpium Citrus Reticulatae, 6-12 parts of Talc, 2-4 parts of Flos Lonicerae, 6-12 parts of Portulaca Oleracea and 1-2 parts of Radix Glycyrrhizae.

[0006] Furthermore, the active ingredients of the Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens are made from the following raw materials in parts by weight: 8.1 parts of Artemisia annua, 16.2 parts of Rhizoma Anemarrhenae, 8.1 parts of Rhizoma Pinelliae, 16.3 parts of Rhizoma Poriae, 8.1 parts of Radix Scutellariae, 8.1 parts of Fructus Aurantii Immaturus, 8.1 parts of Pericarpium Citrus Reticulatae, 10.8 parts of Talc, 3.6 parts of Flos Lonicerae, 10.8 parts of Portulaca Oleracea, and 1.8 parts of Radix Glycyrrhizae.

[0007] Furthermore, the active ingredients of the Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens are made from the following raw materials in parts by weight: 6.5 parts of Artemisia annua, 17 parts of Rhizoma Anemarrhenae, 7 parts of Rhizoma Pinelliae, 18 parts of Poria cocos, 6 parts of Scutellaria baicalensis, 7 parts of Fructus Aurantii Immaturus, 8.5 parts of Citrus reticulatae, 12 parts of Talcum, 4 parts of Flos Lonicerae, 12 parts of Portulaca Oleracea, and 2 parts of Licorice.

[0008] The present invention also provides a method for preparing the above-mentioned Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens, comprising the following steps:

[0009] (1) Weigh the above raw materials in parts by weight, mix them and place them in a Chinese medicine extraction tank, add water and boil them twice, the specific method is: add 7 times the weight of water for the first time, soak for 1 hour, boil for 1 hour, filter to obtain a primary filtrate; add 3 times the weight of water for the second time, boil for 0.5 hour, filter to obtain a secondary filtrate;

[0010] (2) combining the primary filtrate and the secondary filtrate obtained in step (1) to obtain a combined filtrate; concentrating the combined filtrate under reduced pressure to 1 g crude drug / mL to obtain a concentrated solution;

[0011] (3) Add a preservative to the concentrated solution obtained in step (2), and stir evenly to obtain the product.

[0012] Furthermore, the preservative in step (3) is sodium benzoate accounting for 0.3% by weight of the concentrated solution.

[0013] The present invention also provides the use of the Chinese veterinary oral liquid as a medicine for preventing and treating damp-heat diarrhea in chickens.

[0014] Dosage and Administration: 1 ml / kg body weight per day, drink water, for 4-5 consecutive days.

[0015] The Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens of the present invention is formulated according to the traditional theory of Chinese veterinary medicine, based on the physiological and health conditions of chickens, and the basis for syndrome differentiation and treatment and pharmacological analysis are as follows:

[0016] Damp-heat diarrhea is a disease characterized by abdominal pain, urgent diarrhea, or diarrhea without relief, yellow-brown stool, foul odor, burning sensation in the anus, fever, yellow and greasy tongue coating, and a moist or slippery pulse. Its pathogenesis is that the body is affected by damp-heat evil, which blocks the spleen and stomach, injures the intestines and viscera, and causes the spleen and stomach to raise the clear and lower the turbid, and the intestines and viscera to transmit and transform abnormally. Urgent diarrhea is all due to heat, and there is heat in the intestines, so diarrhea is urgent. Damp-heat is intertwined, and the dampness is sticky, blocking the large intestine's qi, and the discharge is not smooth, so diarrhea is not relieved. Damp-heat descends, so the anus is burning, and the stool is yellow and foul. In severe cases, damp-heat injures the blood vessels in the intestines, and pus and blood may be present in the stool. Fever and thirst, yellow and greasy tongue coating, and a moist or slippery pulse are all signs of damp-heat in the body.

[0017] Treatment for this disease: clear away heat and remove dampness.

[0018] The formula of the present invention is composed of Artemisia annua, Rhizoma Anemarrhenae, Rhizoma Pinelliae, Red Poria, Scutellariae Baicalensis, Fructus Aurantii Immaturus, Tangerine Peel, Talcum, Flos Lonicerae, Portulaca Oleracea, and Liquorice. Artemisia annua is slightly bitter, cold and aromatic, lightly dispels heat, removes filth and turbidity; Scutellariae Baicalensis is bitter and cold, and clears away the stagnant heat in the gallbladder and stomach. The two are matched, which can not only expel evil spirits and go out, but also clear dampness and heat inside, and are the main medicines. Rhizoma Anemarrhenae and Rhizoma Pinelliae can clear away phlegm and heat and stop vomiting; Tangerine Peel and Fructus Aurantii Immaturus can regulate qi, relieve stagnation and relieve depression, harmonize the stomach and reduce adverse reactions, and are the assistant medicines. Red Poria can invigorate the spleen, penetrate and eliminate dampness; Talcum and Liquorice can guide dampness and heat downward. Flos Lonicerae and Portulaca Oleracea can clear away heat and detoxify, cool blood and stop dysentery. All the medicines are used together to play the functions of clearing heat, removing dampness, harmonizing the stomach and stopping diarrhea. It has a good therapeutic effect on damp-heat diarrhea caused by external damp-heat evil spirits. It can be used for the prevention and treatment of damp-heat diarrhea in chickens, with good efficacy and no drug residue. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0020] Figure 1 The average body temperature change curve of each group on different days after the virus attack in Test Example 1 of the present invention;

[0021] Figure 2 The histopathological observation results of the duodenal mucosa tissue of the control drug group after the challenge of the test example 1 of the present invention;

[0022] Figure 3 The results of histopathological observation of duodenal mucosa tissue of the test drug group after challenge with poison in Test Example 1 of the present invention;

[0023] Figure 4 The results of histopathological observation of duodenal mucosa tissue of the positive control group after challenge with toxic substances in Test Example 1 of the present invention;

[0024] Figure 5 The results of histopathological observation of duodenal mucosa tissue in the negative control group after challenge with toxic substances in Test Example 1 of the present invention;

[0025] Figure 6 This is a diagram of clinical symptoms and autopsy lesion results of Case 1 in the application example of the present invention.

[0026] in, Figure 2-Figure 5 The middle and right images are enlarged views of their corresponding left images. DETAILED DESCRIPTION

[0027] The present invention is further described below in conjunction with specific embodiments, but the protection scope of the present application is not limited to these embodiments.

[0028] Embodiment 1:

[0029] The Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens is prepared from the following raw materials by weight:

[0030] Artemisia annua 8.1kg, Rhizoma Anemarrhenae 16.2kg, Pinellia ternata 8.1kg, Poria cocos 16.3kg, Scutellaria baicalensis 8.1kg, Fructus Aurantii Immaturus 8.1kg, Citrus reticulatae 8.1kg, Talc 10.8kg, Flos Lonicerae 3.6kg, Portulaca oleracea 10.8kg, Root of Licorice 1.8kg.

[0031] The preparation method of the Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens comprises the following steps:

[0032] (1) Weigh the above-mentioned raw materials (a total of 100 kg), mix them and put them into a Chinese medicine extraction tank, add water (7 times the total weight of the Chinese medicine raw materials, i.e. 700 kg), soak for 1 hour, boil, keep boiling for 1 hour, drain the liquid, and filter to obtain a primary filtrate; add water (3 times the total weight of the Chinese medicine raw materials, i.e. 300 kg), boil, keep boiling for 0.5 hour, drain the liquid, and filter to obtain a secondary filtrate;

[0033] (2) combining the primary filtrate and the secondary filtrate obtained in step (1) to obtain a combined filtrate, placing the combined filtrate in a high-efficiency concentrator and concentrating the combined filtrate under reduced pressure until each milliliter of the concentrated solution is equivalent to 1 g of the crude drug (i.e., 100 liters), thereby obtaining a concentrated solution;

[0034] (3) adding 0.3% sodium benzoate by weight to the concentrated solution obtained in step (2) for preservation, stirring evenly and filling into 500 ml medical plastic bottles to obtain the Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens.

[0035] Dosage and Administration: 1 ml / kg body weight per day, drink water, for 4-5 consecutive days.

[0036] Embodiment 2:

[0037] The Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens is prepared from the following raw materials by weight:

[0038] Artemisia annua 6.5kg, Rhizoma Anemarrhenae 17kg, Pinellia ternata 7kg, Poria cocos 18kg, Scutellaria baicalensis 6kg, Fructus Aurantii Immaturus 7kg, Citrus reticulatae 8.5kg, Talc 12kg, Honeysuckle 4kg, Portulaca oleracea 12kg, Licorice 2kg.

[0039] The preparation steps and dosage are as in Example 1.

[0040] In order to further verify the efficacy of the oral liquid of the present invention in preventing and treating diarrhea caused by damp heat in chickens, the commonly used Gegenqinlian decoction was selected as a control drug to conduct a clinical efficacy test.

[0041] 1. Test Example 1

[0042] 1. Test materials

[0043] 1.1 Traditional Chinese Medicine

[0044] Test drug: oral solution prepared in Example 1 of the present invention, with a content of 1 g of crude drug per mL.

[0045] Control drug: Gegenqinlian decoction, drug composition: 15g of Pueraria root, 9g of Coptis chinensis, 6g of Licorice root, 9g of Scutellaria baicalensis. All Chinese herbal medicine slices were purchased from Jinan Baiweitang Chinese Herbal Medicine Slice Co., Ltd.

[0046] 1.2 Experimental animals: 30-day-old SPF chickens, provided by Shandong Haotai Experimental Animal Co., Ltd.

[0047] 1.3 Escherichia coli strain: strain collection number CVCC1558, purchased from China Veterinary Microbiological Culture Collection Center.

[0048] 1.4 Electronic thermometer: Gush soft-head digital thermometer, accuracy ±0.1℃, produced by Gush Group of Denmark.

[0049] 1.5 Others: Microscope, produced by Olympus Corporation, Japan; ordinary nutrient broth culture medium, produced by Beijing Aoboxing Biotechnology Co., Ltd., batch number 20221103. Blood cell counting chamber, blood collection tube, and saliva aspirator tube were all purchased from Jinan Shengwei Biotechnology Co., Ltd.

[0050] 2 Test methods

[0051] 2.1 Experimental groups: There are 4 groups in total, namely test drug group, control drug group, positive control group and negative control group, with 10 chickens in each group.

[0052] 2.2 Preparation of control drug Chinese medicine solution: According to the composition ratio of Gegenqinlian Decoction, weigh the decoction pieces of each medicinal flavor respectively, add water and boil twice, combine the two decoctions, concentrate to each mL equivalent to 1g of crude drug, and add 1% sodium benzoate for preservation.

[0053] 2.3 Escherichia coli culture: Take 0.2 mL of the preserved bacteria and inoculate it into 50 mL of nutrient broth. Culture it in a shaking incubator at 37°C for 18 h. The concentration of the culture solution is determined to be 1.1 × 10 9 / mL, store at 4℃ for future use.

[0054] 2.4 Virus challenge: Each chicken in the test drug group, control drug group and positive control group was gavaged with 4 mL of the above-mentioned fresh broth culture of Escherichia coli through a gastric tube. The negative control group was not challenged with virus, but was gavaged with 4 mL of drinking water.

[0055] 2.5 Medication method and dosage: 6 hours after the challenge, the test drug group and the control drug group began to take the drug by oral gavage, once in the morning and afternoon every day, 1 ml each time, for 5 consecutive days. The positive control group and the negative control group did not take the drug, but took the same dose of drinking water.

[0056] 2.6 Observation of mental state: Carefully observe the mental state of the chickens every day. If there are symptoms such as feathers standing up, head shrinking and eyes closed, standing still, etc., they are judged to be depressed. If there are no obvious abnormal mental performance, lively expression, eating and drinking normally, they are judged to be normal. Count the number of depressed chicks and the number of normal chicks in each group every day, and calculate the percentage of normal chicks in each group to the total number of chickens.

[0057] 2.7 Observation of feces: The feces of each group of chickens were observed every day, and the feces condition of each group was graded and scored, with a total of 4 levels: watery feces, severe diarrhea, scored 3 points; yellow and thin feces without shape, moderate diarrhea, scored 2 points; yellow and soft feces, basically formed, mild diarrhea, scored 1 point; dark gray feces, formed, with a little white urate attached, normal feces, scored 0 points.

[0058] 2.8 Body temperature measurement: Use an electronic thermometer to measure rectal temperature. Measure body temperature once before challenge and record basal body temperature. After challenge, measure body temperature once at the same time every day and record it until the end of the experiment.

[0059] 2.9 Body weight measurement: Weigh the animals once before the challenge and record the initial body weight; weigh once at the end of the medication; weigh once at the end of the experiment and calculate the weight gain rate.

[0060]

[0061] 2.10 Hematological examination: At the end of the experiment, blood was collected from each chicken wing vein and placed in an EDTA-K2 anticoagulant blood collection tube. The red blood cell count was performed using a counting plate microscope.

[0062] Red blood cell counting: draw 3.98mL of red blood cell diluent into a test tube. Use a Salley pipette to draw the whole blood sample to the 20μL mark. Wipe off the blood adhering to the outer wall of the pipette, blow the blood into the bottom of the test tube, and then blow and aspirate several times to wash the blood cells adhering to the pipette, and then invert the test tube several times to mix. Use a capillary pipette to draw the diluted blood, and place it in the contact point between the counting chamber and the coverslip, let the diluted blood flow naturally into the counting chamber, and let it stand for 1 to 2 minutes. First use a low-power microscope, and the light should be slightly dim. After finding the grid of the counting chamber, put the large square in the center into the field of view, and then switch to a high-power microscope. Select the four corners and the five middle squares in the central large square, count the total number of red blood cells in the five squares, and use the formula X / 80×400×200×10×10 6 Calculate the number of red blood cells in 1L of blood, where X is the total number of red blood cells in the five squares.

[0063] Leukocyte counting method: Place a small drop of blood on a glass slide, make a 30-degree angle between the slide and the glass slide, and make a uniformly distributed thin blood film at a uniform speed. After Wright staining, observe under a microscope. Under the oil immersion lens, select 6 different fields of view from left to right, save the pictures in the computer, and count the red blood cells and white blood cells in the saved pictures in the image J software to calculate the percentage of white blood cells to red blood cells. Calculate the number of white blood cells in the blood of the test chicken by multiplying the total number of red blood cells by the percentage of white blood cells to red blood cells.

[0064] 2.11 Autopsy: At the end of the experiment, all the experimental chickens were bled out from the neck and killed, and the internal organs were dissected and the lesions were recorded.

[0065] 2.12 Histopathological observation of small intestinal mucosa: The duodenum of the chickens was obtained and fixed, stained, wax-sealed, sliced, and examined under a microscope to observe the histopathological changes.

[0066] 3 Test results

[0067] 3.1 Mental status of chickens

[0068] The day after the attack, it was found that all the chickens in the attack group were in a cluster, and some chickens showed obvious symptoms of depression such as shrinking their heads, closing their eyes, feathers standing up, and standing still. No deaths occurred in any group during the entire test period. The mental state of the chickens in the test drug group was significantly better than that in the positive control group, with a significant difference (P<0.05). The mental state of the chickens in the test drug group was also better than that in the control drug group. The negative control group was mentally normal. Detailed results are shown in Table 1.

[0069] Table 1 Statistics of the number of normal chickens in each group at different days after challenge (10 chickens / group)

[0070]

[0071] Note: Data in the same column with different lowercase letters on shoulders indicate significant differences (P<0.05); data with the same lowercase letters on shoulders indicate no significant differences (P>0.05).

[0072] 3.2 Diarrhea

[0073] The day after the attack, it was found that all the chickens in the attack group showed diarrhea symptoms, with yellow loose feces. The feces of each group and the scores of diarrhea severity are shown in Table 2 and Table 3. The experiment showed that the degree of diarrhea in the test drug group and the control drug group was significantly lighter from the fourth day of medication. According to the diarrhea severity score, the diarrhea severity in the test drug group was significantly lighter than that in the positive control group, with a significant difference (P<0.05); the diarrhea severity in the test drug group was also significantly lighter than that in the control drug group, with a significant difference (P<0.05). The feces of the negative control group were normal.

[0074] Table 2 Statistical table of diarrhea severity in each group at different days after challenge

[0075]

[0076] Table 3 Scoring table of diarrhea severity in each group at different days after challenge

[0077]

[0078] Note: Data in the same column with different lowercase letters on shoulders indicate significant differences (P<0.05); data with the same lowercase letters on shoulders indicate no significant differences (P>0.05).

[0079] 3.3 Body temperature measurement results

[0080] There was no difference in basal body temperature among the groups before the challenge. After the challenge, fever occurred in the test drug group, the control drug group, and the positive control group. From the first day to the ninth day after the challenge, the average body temperature of the test drug group and the control drug group was lower than that of the positive control group. Starting from the fifth day after medication, the average body temperature of the test drug group was significantly lower than that of the positive control group, and the difference was significant (P<0.05). The average body temperature of each group on different days after the challenge is shown in Table 4, and the average body temperature change curve of each group on different days after the challenge is shown in Figure 1 The test results showed that the tested drugs had a good heat-clearing effect.

[0081] Table 4 Average body temperature of each group on different days after challenge

[0082]

[0083]

[0084] Note: Data in the same row with different lowercase letters on the shoulders indicate significant differences (P<0.05); data with the same letters on the shoulders or no letters on the shoulders indicate no significant differences (P>0.05).

[0085] 3.4 Weight gain: The weighing and weight gain rate of each group on the 6th and 9th days after infection are shown in Table 5. The results show that the weight gain rate of the test drug group on the 6th and 9th days was higher than that of the positive control group and the control drug group, and the weight gain rate on the 9th day even exceeded that of the negative control group.

[0086] Table 5 Weight and weight gain rate of each group

[0087]

[0088] 3.5 Hematological examination results

[0089] The results of red blood cell and white blood cell counts are shown in Table 6. The results showed that there was no significant difference in red blood cell counts among the groups. The white blood cell counts of the test chickens in the test drug group and the control drug group were lower than those in the positive control group and higher than those in the negative control group. An increase in the number of white blood cells generally indicates a manifestation of bacterial inflammation, indicating that the inflammation in the test drug group was lighter than that in the positive control group.

[0090] Table 6 Average blood leukocyte count results

[0091]

[0092] 3.6 Autopsy results

[0093] Autopsy showed that some chicks in the positive control group had mild pericarditis and perihepatitis, while the test drug group and the control drug group did not have pericarditis, perihepatitis, or peritonitis. All the test chickens in the test drug group, the control drug group, and the positive control group showed enteritis lesions, intestinal mucosal shedding, intestinal wall thinning, thin and yellow contents, but the intestinal mucosa shedding in the positive control group was the most serious, the intestinal wall was thinner, and 2 other chickens had intestinal mucosal bleeding. The enteritis lesions in the test drug group were the mildest, with no mucosal bleeding changes. There were no obvious lesions in the negative control group. The statistics of lesions are shown in Table 7.

[0094] Table 7 Number of chicks with different lesions in each group (10 chicks / group)

[0095]

[0096] 3.7 Pathological observation of small intestinal mucosa

[0097] In the test drug group, control drug group, and positive control group, extensive shedding of intestinal villus epithelial cells was observed in the duodenal tissue mucosa layer of chicks, loose arrangement of intestinal glands in the lamina propria, and scattered inflammatory cell infiltration. However, compared with the positive control group, the pathological changes of the small intestinal mucosal tissue in the test drug group and control drug group were milder; in the negative control group, long and abundant intestinal villi were observed in the duodenal tissue mucosa layer, and abundant and neatly arranged intestinal glands in the lamina propria, without shedding and obvious pathological changes. For details of the histopathological changes in each group, see Figure 2-Figure 5 .

[0098] See also Figure 2 The duodenal tissue of the control drug group was composed of four layers: mucosa, submucosa, muscularis and serosa; the mucosa layer was composed of epithelium, lamina propria and muscularis mucosa; there were many villi on the surface of the mucosa, the epithelium was a single-layer columnar epithelium, and a large amount of mucosal epithelial shedding could be seen; the lamina propria was a dense connective tissue with a large number of intestinal glands; the muscularis was composed of inner circular muscle and outer circular muscle; the serosa was composed of connective tissue and mesothelium; the structure of each layer was clear, and no obvious pathological changes were observed.

[0099] join Figure 3 In the duodenal tissue of the test drug group, a large amount of intestinal villi mucosal epithelium was seen to be shed. The intestinal villi were arranged irregularly, of different lengths, and had unclear structures. The lamina propria was exposed, separated from the mucosal epithelium, and a small amount of intestinal villi and intestinal glands shed into the intestinal cavity.

[0100] See also Figure 4 In the duodenal tissue of the positive control group, a large amount of intestinal villus mucosal epithelium was seen to be shed, the intestinal villi were arranged irregularly, of different lengths, and the structure was unclear; the lamina propria was exposed, and a small amount of lamina propria was seen to be fused with each other, and the structure was blurred; the number of intestinal glands decreased, the structure disappeared, and the structure of some intestinal glands was incomplete and missing.

[0101] See also Figure 5 The duodenal tissue of the negative control group was composed of four layers: mucosa, submucosa, muscularis and serosa; the mucosa layer was composed of epithelium, lamina propria and muscularis mucosa; there were many villi on the surface of the mucosa, and the epithelium was a single-layer columnar epithelium; the lamina propria was a dense connective tissue with a large number of intestinal glands; the muscularis was composed of inner circular muscle and outer circular muscle; the serosa was composed of connective tissue and mesothelium; the structure of each layer was clear, and no obvious pathological changes were observed.

[0102] The above results show that the chickens in the control drug group, test drug group and positive control group all showed intestinal mucosal damage, but the degree of small intestinal mucosal damage in the test drug group was milder than that in the positive control group.

[0103] 4 Conclusion

[0104] From the above test results, the tested drug group showed obvious therapeutic effects in terms of mental state, degree of diarrhea, temperature change, white blood cell count, weight gain rate, and pathological changes compared with the positive control group, and the difference was significant (P<0.05). Compared with the control drug Gegenqinlian decoction, the tested drug group showed better performance in mental state, degree of diarrhea, temperature change, white blood cell count, weight gain rate, and pathological changes. Comprehensive various drug efficacy evaluation indicators show that the oral liquid of the present invention has a good therapeutic effect on chicken damp-heat diarrhea, and the therapeutic effect is better than the traditional drug Gegenqinlian decoction.

[0105] 2. Test Example 2:

[0106] The following test example 2 is used to verify the preventive and therapeutic effect of the oral liquid of the present invention on chicken damp-heat diarrhea:

[0107] 1 Test materials

[0108] 1.1 Drugs under investigation

[0109] Control drug: Dayuan Drink, the medicinal composition is: Areca nut 6g, Magnolia officinalis 3g, Amomum villosum seed 1.5g, Anemarrhena asphodeloides 3g, Paeonia lactiflora 3g, Scutellaria baicalensis 3g, Licorice 1.5g.

[0110] Test drug: oral solution prepared in Example 1 of the present invention.

[0111] 1.2 Experimental animals: SPF chicks, 24 days old, 32 in total.

[0112] 1.3 Escherichia coli strain: Standard Escherichia coli, strain collection number CVCC1558, can cause dysentery in chickens, can be transmitted through the digestive tract, drinking water, feed, etc., and can cause death of chickens. Bacterial content 1.126×10 9 / ml.

[0113] 2 Test methods

[0114] 2.1 Experimental grouping: The experiment was divided into 4 groups: test drug group, control drug group, positive control group and negative control group, with 8 chickens in each group, and each chicken was numbered.

[0115] 2.2 Virus challenge: Each chicken in the test drug group, control drug group, and positive control group was gavaged with 4 ml of E. coli broth culture solution. The negative control group was not challenged with virus.

[0116] 2.3 Medication method and dosage: The control drug group was gavaged with Dayuanyin, and the test drug group was gavaged with the oral liquid of the present invention. Each chicken was orally gavaged with 1 ml once, once in the morning and afternoon every day, for 6 consecutive days.

[0117] 2.4 Clinical observation: Clinical observations were made and recorded for each chicken in the morning and afternoon every day, including head retraction, eye closure, standing still, feathers standing up, mental depression, paralysis, death, etc. The feces of each chicken were observed and recorded. According to the feces condition, there were 4 standards for symptom scoring: watery diarrhea, flowing, symptom score was 6 points; thin feces, unformed, score was 4 points; thin feces, some formed, score was 2 points; normal feces, formed and relatively dry, score was 0 points).

[0118] 2.5 Body temperature measurement: Measure body temperature once before the challenge and record the basal body temperature. After the challenge, measure body temperature again in the afternoon of the same day. Measure body temperature once a day and record it until the end of the experiment. Use an electronic thermometer to measure rectal temperature. Measure by the same person with the same thermometer.

[0119] 2.6 Body weight measurement: Weigh the body weight once before the challenge and record the basal body weight; weigh once at the end of the medication; and weigh once at the end of the experiment.

[0120] 2.7 Autopsy: All dead chickens were autopsied. At the end of the experiment, all experimental chickens were slaughtered, autopsied, and the lesions were recorded.

[0121] 3 Test results:

[0122] 3.1 Mental status

[0123] The day after the challenge, it was found that all the chickens in the challenge group showed slight mental depression, but no serious symptoms of head shrinkage or freezing were found, and no deaths occurred in any group during the entire test period. In terms of mental state, the mental state of the chickens in the test drug group was better than that of the positive control group. The mental state of the chickens in the negative control group was normal.

[0124] 3.2 Stool

[0125] On the second day after the infection, each infection group began to experience diarrhea symptoms. There was no obvious difference between the two drug groups and the positive control.

[0126] 2 days after the challenge, all challenge groups showed diarrhea symptoms. Overall, the degree of diarrhea in the positive control group was significantly more serious than that in the two drug groups. The positive control group had yellow thin feces, most of which were unformed; the feces in the two drug groups were mostly loose and soft, some were formed, and some were normal feces, and the degree of diarrhea was significantly lighter than that in the positive control group; the negative control group had normal feces, which were formed, dry, with white urate on them, and a small amount of loose and slightly formed feces.

[0127] 3-4 days after the challenge, all challenge groups showed diarrhea. From the perspective of feces, the test drug group was the best, followed by the control drug group, and the positive control group was the worst. The negative control group was normal.

[0128] After 5 days of infection, the stool condition had improved significantly, but overall, the best to worst were the negative control group, the test drug group, the control drug group, and the positive control group. Compared with the infection control group, the diarrhea in the test drug group was significantly lighter.

[0129] In summary, from the perspective of diarrhea, both drug groups showed good antidiarrheal effects, with the test drug group being the best and the control drug group being the second best, both of which showed significant differences compared with the challenge control group. The diarrhea scores of each group per day are shown in Table 8.

[0130] Table 8 Diarrhea scores of each group on different days after challenge

[0131]

[0132] Note: Average score, data with different lowercase letters on the shoulders of the same row indicate significant differences (P<0.05); data with the same lowercase letters on the shoulders indicate no significant differences (P>0.05).

[0133] 3.2 Body temperature measurement results

[0134] From the results of body temperature measurement, no fever symptoms appeared in the challenge groups of this experiment, and there was no significant difference compared with the negative control group. The results are shown in Table 9.

[0135] Table 9 Body temperature of each group on different days after challenge (℃)

[0136]

[0137] 3.3 Weight gain results are shown in Table 10 below.

[0138] Table 10 Weight gain statistics of each group

[0139]

[0140] From the weight gain rate on the 5th and 8th days after the challenge, the test drug group had a good weight gain effect, which was significantly higher than the positive control group and the control drug group. The weight gain of the test drug group was also higher than that of the negative control group. The experiment showed that the test drug had a good growth-promoting effect.

[0141] 3.4 Necropsy lesions

[0142] On the 8th day after the challenge, all the test chickens were killed and the internal organs were pathologically examined. The autopsy results showed that one chicken in the positive control group had mild perihepatitis, and all other chickens had no perihepatitis, pericarditis, or air sac inflammation. All chickens had typical diffuse enteritis lesions, which were specifically manifested as shedding of the intestinal mucosa, thinning of the intestinal wall, thin yellow intestinal contents, and some had hemorrhagic enteritis. From the perspective of lesions, the degree of intestinal mucosal lesions in the test drug group was milder than that in the positive control group.

[0143] 4 Experimental conclusion

[0144] From the perspective of mental state, feces, and autopsy lesions, the tested drugs all showed good therapeutic effects and were better than the control drugs. In particular, the tested drugs showed obvious antidiarrhea effects and growth-promoting effects. The above test results once again show that the Chinese veterinary oral liquid of the present invention has a good therapeutic effect on damp-heat diarrhea in chickens.

[0145] In this experiment, none of the test chickens showed fever, which may be related to the fact that the strain was stored for too long and its virulence decreased. Therefore, the heat-clearing effect of the test drug could not be verified.

[0146] 3. Application Examples

[0147] Case 1:

[0148] At a farm in Linyi, more than 6,000 Hailan brown laying hens were raised in one chicken house. At 190 days of age, the chickens began to have diarrhea, with thin, yellow-green feces and a slight decrease in feed intake. The autopsy showed that the intestinal mucosa had fallen off, the intestinal wall had become thinner, and the intestinal contents were yellow-brown. Figure 6 According to clinical symptoms and autopsy lesions, the syndrome was diagnosed as damp-heat diarrhea. The oral liquid of the present invention was used for treatment, 1 mL per chicken per day, twice a day in the morning and afternoon, for 5 consecutive days.

[0149] Three days after medication, the symptoms of the flock began to ease and the feed intake increased significantly. On the seventh day after medication, the flock completely recovered to normal.

[0150] Case 2:

[0151] A farm in Dezhou, which raised more than 10,000 AA white-feathered broilers, began to fall ill at 25 days of age. The chickens were listless, their feed intake decreased, and they had yellow-brown loose feces. The symptoms had been present for 3 days and were diagnosed as enterotoxic syndrome. According to the theory of traditional Chinese medicine, it could be diagnosed as damp-heat diarrhea. The oral liquid of the present invention was then used for treatment, 1 ml per chicken per day, and the medication was concentrated in drinking water twice in the morning and afternoon for 5 consecutive days.

[0152] After 2 days of medication, his spirit returned to normal, his food intake began to increase, and his diarrhea became significantly lighter. After 5 days of medication, his feces completely returned to normal.

[0153] The above is only the embodiment of the present application, and the protection scope of the present application is not limited by these specific embodiments, but is determined by the claims of the present application. For those skilled in the art, the present application can have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the technical ideas and principles of the present application should be included in the protection scope of the present application.

Claims

1. A Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens, characterized in that: The active ingredients are prepared from the following raw materials in parts by weight: 4.5-9 parts of Artemisia annua, 9-18 parts of Rhizoma Anemarrhenae, 4.5-9 parts of Rhizoma Pinelliae, 9-18 parts of Rhizoma Poriae, 4.5-9 parts of Radix Scutellariae, 4.5-9 parts of Fructus Aurantii Immaturus, 4.5-9 parts of Pericarpium Citrus, 4.5-9 parts of Pericarpium Citrus, 6-12 parts of Talc, 2-4 parts of Flos Lonicerae, 6-12 parts of Portulaca Oleracea and 1-2 parts of Radix Glycyrrhizae.

2. The Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens according to claim 1, characterized in that: The active ingredients are prepared from the following raw materials in parts by weight: 8.1 parts of Artemisia annua, 16.2 parts of Rhizoma Anemarrhenae, 8.1 parts of Rhizoma Pinelliae, 16.3 parts of Rhizoma Poriae, 8.1 parts of Radix Scutellariae, 8.1 parts of Fructus Aurantii Immaturus, 8.1 parts of Pericarpium Citrus Reticulatae, 10.8 parts of Talc, 3.6 parts of Flos Lonicerae, 10.8 parts of Herba Portulacae Oleraceae and 1.8 parts of Radix Glycyrrhizae.

3. The Chinese veterinary oral liquid for preventing and treating damp-heat diarrhea in chickens according to claim 1, characterized in that: The active ingredients are prepared from the following raw materials in parts by weight: 6.5 parts of Artemisia annua, 17 parts of Rhizoma Anemarrhenae, 7 parts of Rhizoma Pinelliae, 18 parts of Poria cocos, 6 parts of Scutellaria baicalensis, 7 parts of Fructus Aurantii Immaturus, 8.5 parts of Citrus reticulatae, 12 parts of Talc, 4 parts of Flos Lonicerae, 12 parts of Portulaca Oleracea and 2 parts of Radix Glycyrrhizae.

4. A method for preparing the Chinese veterinary oral liquid according to any one of claims 1 to 3, characterized in that: The steps are as follows: (1) Weigh the above raw materials in parts by weight, mix them and place them in a Chinese medicine extraction tank, add water and boil them twice, the specific method is: add 7 times the weight of water for the first time, soak for 1 hour, boil for 1 hour, filter to obtain a primary filtrate; add 3 times the weight of water for the second time, boil for 0.5 hour, filter to obtain a secondary filtrate; (2) combining the primary filtrate and the secondary filtrate obtained in step (1) to obtain a combined filtrate; concentrating the combined filtrate under reduced pressure until each milliliter of concentrated solution is equivalent to 1 g of the crude drug, to obtain a concentrated solution; (3) Add a preservative to the concentrated solution obtained in step (2), and stir evenly to obtain the product.

5. The method according to claim 4, characterized in that The preservative in step (3) is sodium benzoate accounting for 0.3% by weight of the concentrated solution.

6. Use of the Chinese veterinary oral liquid as claimed in any one of claims 1 to 3 as a drug for preventing and treating damp-heat diarrhea in chickens.

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