CN117298182A - Anticoccidial traditional Chinese medicine composition and preparation method thereof - Google Patents
Anticoccidial traditional Chinese medicine composition and preparation method thereof Download PDFInfo
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- CN117298182A CN117298182A CN202311386302.2A CN202311386302A CN117298182A CN 117298182 A CN117298182 A CN 117298182A CN 202311386302 A CN202311386302 A CN 202311386302A CN 117298182 A CN117298182 A CN 117298182A
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Classifications
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- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/076—Poria
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
- A61K36/233—Bupleurum
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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Abstract
The invention belongs to the field of traditional Chinese veterinary medicines, and in particular relates to an anticoccidial traditional Chinese medicine composition and a preparation method thereof. The invention is prepared from the following raw materials in parts by weight: 5 to 15 parts of kuh-seng, 3 to 7 parts of chinaberry bark, 3 to 7 parts of hairyvein agrimony, 1 to 5 parts of bupleurum, 3 to 7 parts of poria cocos, 2 to 6 parts of immature bitter orange, 3 to 7 parts of bighead atractylodes rhizome, 1 to 5 parts of liquorice and 1 to 5 parts of cimicifuga foetida. According to the characteristics of chicken coccidiosis and the compatibility principle of traditional Chinese medicine compositions, the traditional Chinese medicine composition for preventing and treating chicken coccidiosis is designed. Animal experiments prove that: the chicken feed additive has remarkable chicken coccidium resisting effect, has a protective effect on the integrity of chicken intestinal tracts infected by coccidium, can strengthen the immune activity of chickens, protect the health of chicken intestinal tracts, improve the growth performance of chickens, and has practical popularization and application values.
Description
Technical Field
The invention belongs to the field of traditional Chinese veterinary medicines, and in particular relates to an anticoccidial traditional Chinese medicine composition and a preparation method thereof.
Background
Chicken coccidiosis is an intestinal disease caused by eimeria cell parasitic protozoa, has high morbidity and mortality, wherein the chicken in 8-20 days is most susceptible to infection of coccidiosis, and in intensive culture, the morbidity of coccidiosis is 50-70%, and the mortality is 30-80% or even higher. Chicken coccidiosis infection causes reduced growth performance and increased mortality of broilers, and causes huge economic loss to the global poultry farming industry every year.
At present, chicken coccidiosis is mainly prevented and treated by chemical medicaments and vaccines, but due to long-term use of the chemical medicaments, drug resistance appears, so that a plurality of medicaments cannot play a role. And the medicine is used for a long time, so that the problems of medicine residue and the like in animal-derived products occur. The coccidium live vaccine also has the problems of poor stability, easy virulence return, easy uneven immunity and the like, so the existing coccidium live vaccine is not widely applied to broiler chicken breeding. The European Union has completely banned the use of antibiotics in livestock and poultry production in 2006; japan and korea have also prohibited the use of antibiotics as growth promoters in animal feeds in 2008 and 2011, respectively; in 2020, national agricultural authorities officially release announcements about the addition of growth-promoting drugs other than traditional Chinese medicines to feeds which are completely prohibited. The plant-derived anticoccidial drug has great potential in preventing and treating chicken coccidiosis due to safety and effectiveness and no drug resistance.
In recent years, certain progress has been made in preventing and treating chicken coccidiosis by using traditional Chinese medicines or traditional Chinese medicine formula medicines formed according to compatibility, and the traditional Chinese medicines have unique advantages in the aspects of treating the coccidiosis and enhancing immunity. Research shows that the mixture for expelling the ball and stopping dysentery, which takes antifebrile dichroa, pulsatilla root, hairyvein agrimony, purslane and humifuse euphorbia herb as main raw material medicines, has been used for treating chicken coccidiosis; the Chinese herbal compound prepared from the extracts of the kuh-seng, the sweet wormwood and the astragalus has the effects of preventing and treating chicken coccidiosis, and the treatment effect is better than that of the chemical drug chlorothalonil. However, the anticoccidial traditional Chinese medicine preparation is mainly plant superfine powder, which is not beneficial to quality control.
For example, patent CN108210764a discloses a traditional Chinese medicine composition for treating chicken coccidiosis, which has anticoccidial effect, but is still superfine powder of several medicinal plants, lacks quality control, only shows anticoccidial effect, and is unknown whether the growth performance of chicken infected with coccidiosis is improved. Therefore, the development of a method for treating chicken coccidiosis, enhancing immunity, protecting the integrity of chicken intestinal tract infected by coccidium, improving the growth performance of chicken, greatly reducing the influence of coccidium infection on the growth performance of chicken, and simultaneously enabling the quality of products to be easily controlled is necessary by means of modern extraction, separation and purification technology.
Disclosure of Invention
In order to solve the problems, the invention provides an anticoccidial traditional Chinese medicine composition which is prepared from the following raw materials in parts by weight:
5 to 15 parts of kuh-seng, 3 to 7 parts of chinaberry bark, 3 to 7 parts of hairyvein agrimony, 1 to 5 parts of bupleurum, 3 to 7 parts of poria cocos, 2 to 6 parts of immature bitter orange, 3 to 7 parts of bighead atractylodes rhizome, 1 to 5 parts of liquorice and 1 to 5 parts of cimicifuga foetida.
Further, the material is prepared from the following raw materials in parts by weight:
10 parts of kuh-seng, 5 parts of chinaberry bark, 5 parts of hairyvein agrimony, 3 parts of radix bupleuri, 5 parts of poria cocos, 4 parts of immature bitter orange, 5 parts of bighead atractylodes rhizome, 3g of liquorice and 3 parts of cimicifuga foetida.
Further, the bighead atractylodes rhizome is fried bighead atractylodes rhizome; the Glycyrrhrizae radix is radix Glycyrrhizae Preparata.
Further, the preparation is prepared by taking medicinal powder of the raw material medicine or water or organic solvent extract of the raw material medicine as an active ingredient and adding pharmaceutically acceptable auxiliary materials.
Still further, the formulation is an oral formulation.
Further, the oral preparation is granule, paste, powder, pill or solution.
Further, the total sugar content in the extract is 45-46%, the total protein content is 0.41-0.44%, the matrine content is 0.6-0.8%, the oxymatrine content is 0.06-0.08%, the glycyrrhizin content is 0.13-0.15%, the isoferulic acid content is 0.2-0.3%, the quercetin content is 0.006-0.007%, the toosendanin content is 0.02-0.03%, the glycyrrhizic acid content is 0.22-0.24%, the saikosaponin A content is 0.12-0.13%, and the saikosaponin D content is 0.09-0.11%; the HPLC fingerprint of the extract is shown in figure 1.
Further, the HPLC fingerprint of the extract is detected under the chromatographic conditions of matrine, oxymatrine, glycyrrhizin, glycyrrhizic acid, isoferulic acid, quercetin, toosendanin, saikoside A and saikoside D contents:
chromatographic column: inerSustin-C18 column (250X 4.6mm,5 μm);
mobile phase: acetonitrile (a) -0.1% phosphoric acid water (B);
gradient elution: 0-10 min, 5-4%A; 10-20 min, 4-5%A; 20-30 min, 5-15% of A; 30-60 min, 15-19% of A; 60-95 min, 19-38% of A; 95-120 min, 38-61% A;
detection wavelength 215nm; column temperature 40 ℃; the sample injection amount is 10 mu L; the flow rate was 0.8mL/min.
The invention also provides a preparation method of the pharmaceutical composition, which comprises the following steps:
(1) Weighing the raw materials according to the proportion;
(2) Soaking the raw materials in water, extracting, precipitating the extractive solution with ethanol, filtering, concentrating the filtrate, and drying.
Further, the raw materials are soaked in 3-7 times of water for 1-3 hours, then the mixture is subjected to reflux extraction for 1-3 hours, filtration is carried out, 3-7 times of water is added into filter residues for reflux extraction, filtration is carried out, the two filtrates are combined, ethanol is added until the ethanol content reaches 70%, standing is carried out for 12-48 hours, filtration is carried out, and the filtrate is concentrated and freeze-dried, thus obtaining the medicine.
The invention finally provides application of the traditional Chinese medicine composition in preparing an anticoccidial medicine.
Further, the medicament is a medicament for treating chicken coccidiosis.
The invention aims at the problem that coccidian invades the organism and then colonizes in intestinal epithelial cells to proliferate in a large amount, so that intestinal mucosa inflammation can be caused, nutrient absorption is blocked, and listlessness, bloody stool and even death are caused. Therefore, according to the characteristics of chicken coccidiosis and the compatibility principle of the traditional Chinese medicine composition, the traditional Chinese medicine composition for preventing and treating chicken coccidiosis is designed. The kuh-seng and the chinaberry in the composition have the main effects of inhibiting parasitic diseases, and the hairyvein agrimony has the effects of stopping bleeding and stopping diarrhea; bupleurum, tuckahoe, fructus aurantii and bighead atractylodes rhizome have the effects of resisting inflammation and improving immunity; and licorice and cimicifuga rhizome are used in antipyretic and antibacterial auxiliary treatment. Animal experiments prove that: the traditional Chinese medicine composition has remarkable chicken coccidium resisting effect, has a protective effect on the integrity of chicken intestinal tracts infected by coccidium, can strengthen chicken immunocompetence, protect chicken intestinal tracts healthy, and improve chicken growth performance, and simultaneously can control the quality of the traditional Chinese medicine composition through a phenol-sulfuric acid method, a coomassie brilliant blue method and a specific HPLC method, thereby ensuring the stability and consistency of curative effects and having practical popularization and application values.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Drawings
FIG. 1 is a HPLC fingerprint of radix Sophorae Flavescentis Jiujian powder (1-9 in the figure are respectively matrine, oxymatrine, glycyrrhizin, glycyrrhizic acid, isoferulic acid, quercetin, toosendanin, saikoside A, saikoside D);
FIG. 2 shows organ indices (A heart index, B, liver index, C, spleen index, D, kidney index, E, bursa index) of broiler chickens of each treatment group;
FIG. 3 effect on ZO-1, occludin, claudin-1, MUC2 content in cecal tissue (A.ZO-1, B.Occludin, C.Claudin-1, D.MUC2)
Detailed Description
Example 1 preparation of anticoccidial Chinese medicinal composition of the present invention
The formula comprises the following components: 10g of kuh-seng, 5g of chinaberry bark, 5g of hairyvein agrimony, 3g of bupleurum, 5g of poria cocos, 4g of immature bitter orange, 5g of stir-fried bighead atractylodes rhizome, 3g of honey-fried licorice root and 3g of cimicifuga foetida.
The preparation method comprises the following steps: weighing the raw materials according to the proportion, adding distilled water with the feed-liquid ratio of 1:5 for soaking for 2 hours, carrying out reflux extraction for 2 hours, filtering, adding distilled water 5 times of the filter residues, carrying out reflux extraction for 2 hours, filtering to remove impurities, combining the two filtrates, concentrating the filtrate, slowly adding ethanol to ensure that the alcohol content reaches 70%, standing for 24 hours, filtering, concentrating the filtrate, and freeze-drying to obtain the product.
Example 2 preparation of anticoccidial Chinese medicinal composition of the present invention
The formula comprises the following components: 5g of kuh-seng, 3g of chinaberry bark, 3g of hairyvein agrimony, 1g of bupleurum, 3g of poria cocos, 2g of immature bitter orange, 3g of stir-fried bighead atractylodes rhizome, 1g of honey-fried licorice root and 1g of cimicifuga foetida.
The preparation method comprises the following steps: same as in example 1
Example 3 preparation of anticoccidial Chinese medicinal composition of the present invention
The formula comprises the following components: 15g of kuh-seng, 7g of chinaberry bark, 7g of hairyvein agrimony, 5g of bupleurum, 7g of poria cocos, 6g of immature bitter orange, 7g of stir-fried bighead atractylodes rhizome, 5g of honey-fried licorice root and 5g of cimicifuga foetida.
The preparation method comprises the following steps: same as in example 1
The advantageous effects of the present invention are described below by way of test examples.
Experimental example 1 investigation of the composition and efficacy of the anticoccidial Chinese medicinal composition of the present invention
1. Component detection
The total sugar content and protein content in 10 batches of the traditional Chinese medicine extract prepared in example 1 are respectively measured by a phenol-sulfuric acid method and a coomassie brilliant blue method, and simultaneously, a High Performance Liquid Chromatography (HPLC) fingerprint of the extract is established by an HPLC method, so that 9 components such as matrine, oxymatrine, glycyrrhizin, glycyrrhizic acid, isoferulic acid, quercetin, toosendanin, saikosaponin A, saikosaponin D and the like are quantitatively analyzed. The specific HPLC detection method comprises the following steps:
1) Sample solution preparation: taking 0.6g of the traditional Chinese medicine extract freeze-dried product prepared in the example 1, adding 30mL of methanol, weighing the mass, sealing, performing ultrasonic treatment for 30min (ultrasonic frequency 40KHz and ultrasonic power 240W), standing at room temperature, weighing the mass, supplementing the mass with methanol, shaking uniformly, and passing through a 0.45 mu m microporous filter membrane to obtain the traditional Chinese medicine extract freeze-dried product;
2) Preparation of a control solution: precisely weighing matrine reference substance, oxymatrine reference substance, glycyrrhizic acid reference substance, glycyrrhizin reference substance, quercetin reference substance, toosendanin reference substance, isoferulic acid reference substance, saikosaponin A reference substance and saikosaponin D reference substance, and adding methanol to obtain individual reference substance solutions of matrine 0.84mg/mL, oxymatrine 0.20mg/mL, glycyrrhizin 0.60mg/mL, glycyrrhizic acid 0.25mg/mL, isoferulic acid 0.64mg/mL, quercetin 22 μg/mL, toosendanin 0.18mg/mL, saikosaponin A0.40 mg/mL, and saikosaponin D0.45 mg/mL. Precisely weighing matrine reference substance, oxymatrine reference substance, glycyrrhizic acid reference substance, glycyrrhizin reference substance, quercetin reference substance, toosendanin reference substance, isoferulic acid reference substance, saikosaponin A reference substance and saikosaponin D reference substance, adding methanol to obtain mixed reference substance solutions containing matrine 0.084mg/mL, oxymatrine 0.02mg/mL, glycyrrhizin 0.06mg/mL, glycyrrhizic acid 0.05mg/mL, isoferulic acid 0.064mg/mL, quercetin 2.2 μg/mL, toosendanin 0.018mg/mL, saikosaponin A0.04 mg/mL and saikosaponin D0.045 mg/mL.
3) Detecting the sample solution and the reference solution by adopting the following chromatographic conditions, collecting fingerprint patterns and calculating the content of each component;
chromatographic column: inerSustin-C18 column (250X 4.6mm,5 μm);
mobile phase: acetonitrile (a) -0.1% phosphoric acid water (B);
gradient elution: 0-10 min, 5-4%A; 10-20 min, 4-5%A; 20-30 min, 5-15% of A; 30-60 min, 15-19% of A; 60-95 min, 19-38% of A; 95-120 min, 38-61% A;
detection wavelength 215nm; column temperature 40 ℃; the sample injection amount is 10 mu L; the flow rate was 0.8mL/min.
The specific results are shown in tables 1-2, and FIG. 1, the results show that the quality of the extracts of different batches is stable.
Table 1 results of total sugar and total protein content in samples
It can be seen from tables 1 to 2: the traditional Chinese medicine composition comprises 45-46% of total sugar, 0.41-0.44% of total protein, 0.6-0.8% of matrine, 0.06-0.08% of oxymatrine, 0.13-0.15% of glycyrrhizin, 0.2-0.3% of isoferulic acid, 0.006-0.007% of quercetin, 0.02-0.03% of toosendanin, 0.22-0.24% of glycyrrhizic acid, 0.12-0.13% of saikosaponin A and 0.09-0.11% of saikosaponin D in an extract freeze-dried product prepared according to the embodiment 1. The active ingredient content is high.
2. Anticoccidial efficacy assays
2.1 Experimental drugs
Kuh-seng Jiujian san (1 g equivalent to 14.7g crude drug) prepared in example 1; coccidium powder; dekkazuril
2.2 Experimental insect strains
The 14-day-old chicks were inoculated with Eimeria tenella sporulated oocysts 5 by oral feeding×10 4 And each. Blood stool was collected from the infection at 5-8d and sacrificed at 8d all, and cecal content was removed. Purifying and separating coccidian oocysts by a saturated saline floatation method, placing the separated oocysts in a 2.5% potassium dichromate solution for sporulation for 72 hours to obtain a soft Eimer sporulation oocyst suspension, and preserving at 4 ℃ for later use.
2.3 laboratory animals
Three yellow broilers of 1 day old were purchased from the adult dragon-containing poultry hatchery and the broilers were not vaccinated with any vaccine. The feed and the drinking water are fed into a strictly sterilized animal house and a metal cage without any anticoccidial drug.
2.4 grouping and administration
Grouping and administration were as in table 3. The traditional Chinese medicine components are 3 experimental groups: the high dose group 1g/kg, the medium dose group 0.8g/kg and the low dose group 0.6g/kg are all administered by mixing materials. Group 4: the coccidium powder medicine group is mixed with 15g/kg for administration; group 5: the dekkera bead is mixed with the medicine group for administration; group 6, namely a control group without drug for attacking insects; group 7: the control group was not infected with the drug.
Three yellow broilers of 1 day old with uniform body weight were divided into 7 groups of 3 replicates of 12 broilers each. Each group was fed basal feed for 1-14 days. On 14 days, adding 0.6g/kg, 0.8g/kg and 1g/kg of kuh-seng nine-decoct powder into the basic ration of groups 1, 2 and 3 respectively; group 4 adding 15g/kg of coccidium powder to the base ration; group 5 added 0.2g/kg of diclazuril to the basal diet, groups 6 and 7 were fed basal diet. The drug addition group was continuously administered for 7 days. The broilers of the other groups except the 6 th and 7 th groups were orally inoculated with 1×10 at 14 days old 5 Individual eimeria tenella sporulated oocyst suspensions were dissected by day 8 of infection.
Table 3 animal experiment groups and dosing
2.5 measurement index
The weight, feed intake and feed conversion rate of 1 day old, 14 day old and 21 day old broilers were recorded, wherein feed conversion rate = average day feed intake/average day gain.
(1) Relative weight gain (%)
Relative weight gain rate = average weight gain of infected group/average weight gain of non-infected group x 100%
(2) Survival (%)
Survival (%) = (number of chickens per group at the end of the experiment +.1% of chickens per group at the beginning of the experiment)
(3) Efficacy scoring
Oocyst values were scored based on the criteria of angle Tian Qing (1986).
TABLE 4 conversion of oocyst count to oocyst value
The standard of Johnson Reid (1970) scores cecal lesions.
Table 5 cecal lesion scoring criteria. Lesion value = average lesion score per group of chicken cecum x 10 table 5 cecum lesion scoring criteria
The standard of Merck corporation in the united states calculates the anticoccidial index (ACI).
Table 6 anticoccidial index criteria. Aci= (relative rate of weight gain + survival) - (lesion + oocyst value) table 6 anticoccidial index criterion
3. Results
After the 5 th day of infection, the spirit of the non-infected and non-dosed broiler chickens is normal, the feed intake is normal, and no blood and urine appear; the contrast group without using the medicine for attacking the insects has symptoms of mental depression, head and neck contracture, reduced feed intake, diarrhea, bloody stool and the like. The groups treated with the drugs also had anorexia, lassitude, and had different degrees of bloody stool output, but had less blood stool output than the control group without challenge.
The results of the anticoccidial effect evaluation are shown in table 7, and the survival rate of the broilers of each test group is 100%. Except the control group which is not infected and not used, the relative weight gain rate of the broilers in the other test groups is different, which indicates that the growth of the broilers is slowed down due to coccidium infection. The relative weight gain rate of the control group without the drug for attacking insects is 63.32 percent at the lowest. The relative weight gain rates of the traditional Chinese medicine low dose group (0.6 g/kg), the traditional Chinese medicine medium dose group (0.8 g/kg) and the traditional Chinese medicine high dose group (1 g/kg) are 77.83%, 77.60% and 81.77%, respectively. The disease values of the non-infection non-drug control group, the insect attack non-drug control group, the diclazuril group and the coccidium powder are respectively 0, 33.33, 6.67 and 20.83. The cecal lesions of the broiler chickens in the low-dose group (0.6 g/kg), the medium-dose group (0.8 g/kg) and the high-dose group (1 g/kg) of the traditional Chinese medicines are 25.83, 21.67 and 20 respectively. The disease value of cecum of the broiler chickens in the high-dose group (1 g/kg) of the traditional Chinese medicine is higher than that of coccidium powder group and slightly lower than that of the diclazuril group. The coccidian resistance index of the broiler chickens of the traditional Chinese medicine high-dose group (1 g/kg) is 161.10, so that a good coccidian resistance effect is achieved.
The anticoccidial effect of the high-dose group (1 g/kg) of the traditional Chinese medicine reaches a good level. The chemical anticoccidial drug used at present has better efficacy, but can easily generate drug-resistant insect strains after long-term use, so that a plurality of anticoccidial chemical drugs lose the therapeutic effect. The product is mainly composed of traditional Chinese medicines, and is not easy to generate drug resistance and toxic and side effects.
TABLE 7 anticoccidial effect
The organ index results of the broilers of each test group are shown in fig. 2. As can be seen from fig. 2-a, the heart index was significantly increased in the attack model group (P < 0.05) compared to the blank group. Compared with the attack model group, the heart index of each drug addition group is obviously reduced (P is less than 0.05), the heart indexes of the high-dose group, the coccidian group and the diclazuril group of the kuh-seng nine-decoction powder have no obvious difference, and the heart indexes of the different doses of the kuh-seng nine-decoction powder have no obvious difference, but the heart indexes gradually decrease along with the increase of the doses. As can be seen from fig. 2-B, the liver index of the challenged model group broiler chickens was significantly higher than that of the other test groups (P < 0.05), and there was no significant difference between the blank group and the other treatment groups. Compared with the diclazuril group, the liver index of the kuh-seng nine-decoction high-dose group is obviously reduced (P < 0.05). As shown in fig. 2-C, the spleen index was significantly reduced in the challenge model group compared to the blank group (P < 0.05). The spleen index of each group of drug addition is obviously higher than that of the attack model group (P < 0.05), and the spleen index of a blank group, a kuh-seng nine-decoction high-dose group and a diclazuril group has no obvious difference. As shown in fig. 2-D, the kidney index of the attack pattern group was significantly lower than that of the blank group (P < 0.05). Compared with the attack model group, the kidney indexes of the drug addition groups are obviously increased (P < 0.05), wherein the low, medium and high dosage groups of the kuh-seng decoction are obviously higher than the coccidium powder group (P < 0.05). As can be seen from fig. 2-E, the attack model group bursa index was significantly lower than the blank group (P < 0.05). Compared with the attack model group, the bursa of Fabricius index of each group of drug addition is obviously increased (P < 0.05), wherein the medium and high dose groups of kuh-seng are obviously higher than Yu Deke zuli groups and coccidiosis groups (P < 0.05), and no obvious difference exists between the medium and high dose groups and the blank groups.
The results of the cecal tissue tight junction protein and mucin content of each test group are shown in fig. 3. As can be seen from fig. 3-a, compared with the blank group, the content of ZO-1 in the cecum tissue of the meat chicken of the attack model group is significantly reduced (P < 0.05), and the content of ZO-1 in the cecum tissue of the meat chicken treated by the kuh-seng nine-decoct powder dose group, the coccidium powder group and the diclazuril group is significantly higher than that of the meat chicken of the attack model group, and has no significant difference from the blank group. As can be seen from fig. 3-B, the cecal tissue Occludin content of the challenge model group broiler was significantly reduced (P < 0.05) compared to the blank group. Compared with the attack model group, the content of the cecum tissue Occludin of the high-dose group of the kuh-seng and the coccidium powder group is obviously increased (P is less than 0.05), wherein no obvious difference exists between the content of the cecum tissue Occludin of the high-dose group of the kuh-seng and the coccidium powder group. From FIG. 3-C, it can be seen that the content of Claudin-1 in the cecum tissue of the attack model group broiler is significantly reduced (P < 0.05) compared with the blank group. The content of Claudin-1 in the cecum tissue of the kuh-seng Jiujian san, the kuh-seng Jiujian san high dose group, the coccidian san group and the diclazuril group is obviously higher than that of the attack model group, and the kuh-seng Jiujian san medium dose group, the kuh-seng Jiujian san high dose group and the coccidian san group have no obvious difference. As can be seen from fig. 3-D, compared with the blank group, the content of MUC2 in the challenge model group is significantly reduced (P < 0.05), the content of MUC2 in the cecum tissue of the kuh-seng nine-decoct high dose group, the coccidian powder group and the diclazuril group is significantly higher than that in the challenge model group (P < 0.05), and there is no significant difference between the kuh-seng nine-decoct high dose group and the coccidian powder group.
In conclusion, the quality of the traditional Chinese medicine composition is controlled by a phenol-sulfuric acid method, a coomassie brilliant blue method and a specific HPLC method, so that the stability and consistency of curative effects are ensured, animal experiments prove that the traditional Chinese medicine composition has remarkable effects on resisting chicken coccidium, has a protective effect on the integrity of chicken intestinal tracts infected by coccidium, can strengthen the immune activity of chicken, protect the health of chicken intestinal tracts and improve the growth performance of chicken.
Claims (10)
1. An anticoccidial traditional Chinese medicine composition is characterized in that: the traditional Chinese medicine is prepared from the following raw materials in parts by weight:
5 to 15 parts of kuh-seng, 3 to 7 parts of chinaberry bark, 3 to 7 parts of hairyvein agrimony, 1 to 5 parts of bupleurum, 3 to 7 parts of poria cocos, 2 to 6 parts of immature bitter orange, 3 to 7 parts of bighead atractylodes rhizome, 1 to 5 parts of liquorice and 1 to 5 parts of cimicifuga foetida.
2. The traditional Chinese medicine composition according to claim 1, wherein: the composite material is prepared from the following raw materials in parts by weight:
10 parts of kuh-seng, 5 parts of chinaberry bark, 5 parts of hairyvein agrimony, 3 parts of radix bupleuri, 5 parts of poria cocos, 4 parts of immature bitter orange, 5 parts of bighead atractylodes rhizome, 3g of liquorice and 3 parts of cimicifuga foetida.
3. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that: the bighead atractylodes rhizome is fried bighead atractylodes rhizome; the Glycyrrhrizae radix is radix Glycyrrhizae Preparata.
4. The traditional Chinese medicine composition according to claim 1, wherein: the preparation is prepared by taking medicinal powder of raw materials or water or organic solvent extract of the raw materials as an active ingredient and adding pharmaceutically acceptable auxiliary materials; the preparation is an oral preparation; the oral preparation is granule, paste, powder, pill or solution.
5. The traditional Chinese medicine composition according to claim 4, wherein: the total sugar content of the extract is 45-46%, the total protein content is 0.41-0.44%, the matrine content is 0.6-0.8%, the oxymatrine content is 0.06-0.08%, the glycyrrhizin content is 0.13-0.15%, the isoferulic acid content is 0.2-0.3%, the quercetin content is 0.006-0.007%, the toosendanin content is 0.02-0.03%, the glycyrrhizic acid content is 0.22-0.24%, the saikosaponin A content is 0.12-0.13%, and the saikosaponin D content is 0.09-0.11%; the HPLC fingerprint of the extract is shown in figure 1.
6. The traditional Chinese medicine composition according to claim 5, wherein: the HPLC fingerprint of the extract is detected, and chromatographic conditions of the contents of matrine, oxymatrine, glycyrrhizin, glycyrrhizic acid, isoferulic acid, quercetin, toosendanin, saikoside A and saikoside D are as follows:
chromatographic column: inerSustin-C18 column (250X 4.6mm,5 μm);
mobile phase: acetonitrile (a) -0.1% phosphoric acid water (B);
gradient elution: 0-10 min, 5-4%A; 10-20 min, 4-5%A; 20-30 min, 5-15% of A; 30-60 min, 15-19% of A; 60-95 min, 19-38% of A; 95-120 min, 38-61% A;
detection wavelength 215nm; column temperature 40 ℃; the sample injection amount is 10 mu L; the flow rate was 0.8mL/min.
7. A process for the preparation of a pharmaceutical composition as claimed in any one of claims 1 to 6, characterized in that: it comprises the following steps:
(1) Weighing the raw materials according to the proportion;
(2) Soaking the raw materials in water, extracting, precipitating the extractive solution with ethanol, filtering, concentrating the filtrate, and drying.
8. The method of manufacturing according to claim 7, wherein: soaking the raw materials in 3-7 times of water for 1-3 hours, reflux-extracting for 1-3 hours, filtering, reflux-extracting filter residues in 3-7 times of water, filtering, combining the two filtrates, adding ethanol until the ethanol content reaches 70%, standing for 12-48 hours, filtering, concentrating the filtrate, and freeze-drying to obtain the traditional Chinese medicine.
9. Use of the Chinese medicinal composition according to one of claims 1 to 6 for the preparation of an anticoccidial medicament.
10. Use according to claim 9, characterized in that: the medicine is used for treating chicken coccidiosis.
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