CN112933172B - Traditional Chinese medicine compound extract for treating intestinal coccidiosis of rabbits and application thereof - Google Patents

Abstract

The invention discloses a traditional Chinese medicine compound extract for treating rabbit intestinal coccidiosis and application thereof, wherein the raw material medicines of the traditional Chinese medicine compound extract comprise the following components in parts by weight: 40-80 parts of hawthorn, 40-80 parts of antifebrile dichroa, 40-80 parts of betel nut, 40-80 parts of liquorice, 100-150 parts of scutellaria baicalensis, 100-150 parts of coptis chinensis and 100-150 parts of Chinese herbaceous peony. The traditional Chinese medicine compound extract disclosed by the invention can reduce oocyst discharge, effectively treat intestinal coccidiosis of rabbits caused by large-scale Eimeria, medium-scale Eimeria, yellow Eimeria and small-scale Eimeria, and has a wide research value and application prospect in clinic.

Description

Traditional Chinese medicine compound extract for treating rabbit intestinal coccidiosis and application thereof

Technical Field

The invention belongs to the field of traditional Chinese medicine compositions, and particularly relates to a traditional Chinese medicine compound extract for treating rabbit intestinal coccidiosis and application thereof.

Background

Coccidiosis in rabbits is a common and serious infectious disease caused by the parasitism of the coccidia, a variety of unicellular organisms of the genus eimeria, in the epithelial cells of the small intestine or bile duct of rabbits. At present, 7 kinds of eimeria coccidiosis can parasitize the small intestine and the bile duct of rabbits in China. These unicellular organisms belong to the protozoan species and they are most commonly referred to as oocysts. Oocysts, so-called non-sporulating oocysts, are excreted by animals in their faeces. They remain in the environment and form spores. These sporulated oocysts, if ingested, cause diarrhea and sometimes death in the animal. The rabbit coccidian oocysts can develop into infectious oocysts after 2-3 days in an external environment with the temperature of 20 ℃ and the humidity of 55-75%. The resistance of the oocysts to chemical disinfection drugs and low temperature is strong, and most of the oocysts can live through winter. But is sensitive to sunlight and dryness, and direct sunlight can kill oocysts within hours. The disease is susceptible to rabbits of different ages and varieties, wherein young rabbits within 3 months are most susceptible to infection, and the infection rate is up to 100%. If measures are not taken in time after the common rabbit raising field is infected with coccidium, the death rate can reach 70 percent, and great economic loss is caused to the rabbit raising industry.

After coccidiosis infects rabbits, except for liver diseases caused by the coccidiosis, the rest coccidiosis causes damages to intestinal epithelial cells and mucosal tissues of the rabbits, resulting in dehydration, blood loss, diarrhea and may be secondary to other bacterial or viral diseases. At present, although the variety of medicines for treating rabbit coccidiosis is various, the rabbit coccidiosis is still high in habitation mainly because no commercial vaccine exists; in addition, in the production, except for the gestational period, people almost continuously add low-dose anticoccidial drugs into the feed in the whole rabbit raising process, so that not only drugs are not selected in a targeted manner, but also anticoccidial coccidia strains are easily caused by long-term use of one anticoccidial drug, and partial cross resistance exists among the ionophore drugs. In order to better control coccidiosis, the drugs must be exchanged frequently, which also results in a significant increase in production costs. Meanwhile, the toxicity, residue, immunosuppression and drug resistance of chemical drugs seriously troubles the application of the drugs, and seriously affects and harms the health of human beings. Therefore, aiming at the local coccidian strains, the screening of low-toxicity and high-efficiency anticoccidial drugs is carried out, and the method is very important for the targeted prevention and control of rabbit coccidiosis.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine compound extract for treating rabbit intestinal coccidiosis, which can effectively treat rabbit intestinal coccidiosis caused by mixed infection of large Eimeria, medium Eimeria, yellow Eimeria and small Eimeria and can be used for preparing related products for treating and/or preventing rabbit intestinal coccidiosis.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides a traditional Chinese medicine compound extract, which comprises the following raw material medicines in parts by weight: 40-80 parts of hawthorn, 40-80 parts of antifebrile dichroa, 40-80 parts of betel nut, 40-80 parts of liquorice, 100-150 parts of scutellaria baicalensis, 100-150 parts of coptis chinensis and 100-150 parts of Chinese herbaceous peony.

Further, the raw material medicaments comprise the following components in parts by weight: 50-70 parts of hawthorn, 50-70 parts of antifebrile dichroa, 50-70 parts of betel nut, 50-70 parts of liquorice, 110-130 parts of scutellaria baicalensis, 110-130 parts of coptis chinensis and 110-130 parts of Chinese herbaceous peony.

Furthermore, the raw material medicaments comprise the following components in parts by weight: 60 parts of hawthorn, 60 parts of antifebrile dichroa, 60 parts of betel nut, 60 parts of liquorice, 120 parts of scutellaria baicalensis, 120 parts of coptis chinensis and 120 parts of Chinese herbaceous peony.

The invention provides a preparation method of the extract, which comprises the following steps:

(1) taking raw material medicaments in parts by weight; (2) extracting with ethanol or water; (3) mixing, and preparing the product; further, preferably (1) the raw material medicines are taken according to the weight parts; (2) water extraction; (3) mixing, and making into product.

Wherein (1) can be combined with any one or two of (2) and (3), and the raw material medicines in (1) can be respectively and independently selected from hawthorn, dichroa febrifuga, areca nut, liquorice, scutellaria baicalensis, coptis chinensis or peony to be combined with any one or two of (2) and (3), or selected from several of hawthorn, dichroa febrifuga, areca nut, liquorice, scutellaria baicalensis, coptis chinensis and peony to be combined with any one or two of (2) and (3), and the like; the raw material medicines can be directly mixed, or all the raw material medicines are extracted by alcohol or water to prepare extracts for mixing, or a part of the raw material medicines are extracted by alcohol or water, and a part of the raw material medicines are directly extracted by alcohol or water and are mixed.

That is, the following combinations may be included but not limited to:

A. mixing at least two components of each raw material medicine, and then extracting with ethanol or water;

B. extracting each raw material medicine separately and mixing;

for example, mixing fructus crataegi, radix Dichroae, Arecae semen, Glycyrrhrizae radix, Scutellariae radix, Coptidis rhizoma, and radix Paeoniae; or mixing fructus crataegi and radix Dichroae, extracting, and mixing with other raw materials; or mixing fructus crataegi, radix Dichroae and Arecae semen, extracting other materials separately, mixing, etc.

The invention also provides application of the traditional Chinese medicine compound extract in preparing a product for treating and/or preventing coccidiosis.

Further, the product also comprises pharmaceutically acceptable auxiliary materials.

The "pharmaceutically acceptable excipient" of the present invention is meant to include any substance that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered.

The pharmaceutically acceptable auxiliary material is a general name of all additional materials except the main medicine in the medicine, and the auxiliary material has the following properties: (1) no toxic effect on human body and almost no side effect; (2) the chemical property is stable and is not easily influenced by temperature, pH, storage time and the like; (3) has no incompatibility with the main drug, and does not influence the curative effect and quality inspection of the main drug; (4) does not interact with the packaging material.

The auxiliary materials of the invention include, but are not limited to, fillers (diluents), lubricants (glidants or anti-adherents), dispersing agents, wetting agents, binders, regulators, solubilizers, antioxidants, bacteriostats, emulsifiers, disintegrants and the like. The binder comprises syrup, acacia, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethyl cellulose or hydroxypropyl methylcellulose), gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone; the filler comprises lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricant comprises superfine silica gel powder, magnesium stearate, talcum powder, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; the disintegrating agent comprises starch and its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; the wetting agent comprises sodium lauryl sulfate, water or alcohol, etc.; the antioxidant comprises sodium sulfite, sodium bisulfite, sodium pyrosulfite, dibutylbenzoic acid, etc.; the bacteriostatic agent comprises 0.5% of phenol, 0.3% of cresol, 0.5% of chlorobutanol and the like; the regulator comprises hydrochloric acid, citric acid, potassium (sodium) hydroxide, sodium citrate, and buffer (including sodium dihydrogen phosphate and disodium hydrogen phosphate); the emulsifier comprises polysorbate-80, sorbitan fatty acid, pluronic F-68, lecithin, soybean lecithin, etc.; the solubilizer comprises Tween-80, bile, glycerol, etc.

Further, the product is used for preparing a product for treating and/or preventing rabbit intestinal coccidiosis.

Further, the pathogen of rabbit intestinal coccidiosis is selected from Eimeria;

in a specific embodiment of the invention, the Eimeria is selected from one or more of Eimeria maxima, Eimeria necatrix.

Further, the product is an oocyst development inhibitor.

The "oocyst development inhibitor" is a product for inhibiting the development of oocysts.

Further, the product is a product capable of reducing the excretion of oocysts.

The product of the invention can inhibit the endogenous development of the oocysts and reduce the discharge of the oocysts out of the body; help repair mucosal injury, control secondary infection.

The invention has the beneficial effects that: the traditional Chinese medicine compound extract disclosed by the invention exerts the combined effect of dialectical treatment of a traditional Chinese medicine prescription, has an exact curative effect, can be used for preparing related products for treating and/or preventing rabbit intestinal coccidiosis, particularly can effectively inhibit coccidiosis caused by mixed infection of large Eimeria, medium Eimeria, yellow Eimeria and small Eimeria, and has the effects of promoting oocyst discharge, increasing body weight and reducing the death rate of infected rabbits according to the experimental results, meanwhile, the traditional Chinese medicine compound extract disclosed by the invention can repair intestinal mucosa injury and prevent secondary infection, and has a wide research value and application prospect in clinic; the invention has the advantages of rich source of raw material medicines, simple preparation process and low cost, and greatly reduces the production cost.

Drawings

FIG. 1 is a graph of coccidial oocyst expulsion from various groups of rabbits following drug treatment;

FIG. 2 is a diagram of clinical symptoms and intestinal lesions of diseased rabbits;

FIG. 3 is a diagram showing the oocyst shedding of various rabbit coccidia groups from day 3 to day 24 after the attack of the pests.

Wherein, in fig. 2, a: day 4 infected thin manure of sick rabbits, B: severe bleeding of intestinal mucosa of dead rabbits, C: the intestinal tract of the dead rabbit is aerated and bleeds.

Detailed Description

The use of the present invention is further illustrated by the following specific examples and experiments. The following examples are only preferred embodiments of the present invention and are not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

EXAMPLE 1 preparation of herbal Compound extract

Weighing raw material medicines according to 60 parts of hawthorn, 60 parts of antifebrile dichroa, 60 parts of areca, 60 parts of liquorice, 120 parts of scutellaria baicalensis, 120 parts of coptis chinensis and 120 parts of Chinese herbaceous peony, adding distilled water according to 5 times of the total weight of the medicines, carrying out ultrasonic soaking for 1-1.5 hours, starting decoction, setting the decoction temperature at 90-100 ℃, carrying out decoction for 5-6 hours, filtering to obtain a raw medicine filtrate, repeating the above steps for 3 times by using filter residues, and finally combining all the filtrates. Adding 95% ethanol into the obtained filtrate at a ratio of 10:6 for precipitating with ethanol and standing, transferring the supernatant to a rotary evaporator for concentrating when standing for 0.5-1 hr until the volume of the concentrated solution is 1/9 of the volume of the filtrate, and setting the concentration temperature at 60-70 deg.C to obtain the concentrated solution.

Example 2

160 parts of Chinese pulsatilla root, 120 parts of coptis root, 60 parts of antifebrile dichroa and areca seed, 60 parts of malt, medicated leaven and brucea javanica. The extraction method was the same as in example 1.

Example 3

Coptis root 120 g, banksia rose 80 g, areca-nut, white atractylodes rhizome, antifebrile dichroa and areca-nut each 60 g. The extraction method was the same as in example 1.

Test example 1

1 materials

Infectious coccidial oocysts

Clinically, most of rabbit infected coccidia are mixed infection, and in the test, mixed oocysts (large eimeria, medium eimeria, yellow eimeria and small eimeria) collected from excrement of seriously infected coccidiosis rabbits in a certain breeding rabbit farm in Sichuan are selected and hatched into infectious oocysts by potassium dichromate and stored at 4 ℃.

Test drugs

Diclazuril solution was purchased from Hua-beast drugs, Inc. (batch No. 041212045); the 10% robenidine premix was purchased from Zhejiang Hui Neigong Biometrics Ltd (batch No. 110481377); the traditional Chinese medicine compound extracts 1, 2 and 3 are products prepared in the embodiments 1, 2 and 3 respectively.

Anticoccidial test of diclazuril in animal Chinese medicine compound 1, 2, 3 stock solution

Selecting 60 artificial infected intestinal coccidian rabbits (immunized rabbit plague vaccine) of 40 days old, each male and female half, the average body weight is 0.8 kg-1.1 kg, collecting rabbit dung before grouping the experiment, adopting a wheat worm egg counting method to detect oocysts, and enabling the number of the infected intestinal coccidian oocysts to be less than 1 x 10⁴One is mild infection. Each rabbit was fed individually to a sterilized rabbit cage. Weighing one by one, then randomly dividing the weighed mixture into 6 experimental groups, wherein each group comprises 10 groups, namely groups I, II, III, IV, V and VI, wherein 8mL of the Chinese herbal compound extracts of 1, 2 and 3 are respectively irrigated for 7 days for the groups I to III, and 8mL of the Chinese herbal compound extracts of 3 are respectively irrigated for the group IV, and diclazuril solution is added into drinking water according to the concentration of 0.3 mL/L; feeding group V with 600mg/kg chlorobenzeneGuanidine feed, group VI without medication. Before the administration, 1 week after the administration, 2 weeks after the administration, and 3 weeks after the administration, the oocyst excretion, weight gain and mortality of the coccidia of each group of rabbits were counted.

The oocyst shedding changes of the test rabbits are shown in table 1 and fig. 1.

TABLE 1 Rabbit OPG Change in groups in the Eimeria treatment trial

Note: the same data in the same column with the same shoulder mark letters means that the difference is not significant (P >0.05), and the difference means that the difference is significant (P < 0.05).

As can be seen from Table 1 and FIG. 1, each treatment group showed a significant drop in oocysts after administration, with the lowest amount of oocysts at 14 days after administration followed by a gradual increase in amplitude. 7 days, 14 days and 21 days after administration, the difference of the oocyst output of the drug group compared with that of the non-drug group is very obvious (P is less than 0.01); 7 days and 14 days after administration, the oocyst output difference of each treatment group is not significant (P > 0.05); 21 days after administration, the differences of the Chinese medicinal compound 1, the Chinese medicinal compound 2, the Chinese medicinal compound 3 and the robenidine are not significant (P is more than 0.05); the diclazuril group has no significant difference (P is more than 0.05) with the traditional Chinese medicine compound 1, the traditional Chinese medicine compound 2 and the robenidine group, and has significant difference (P is less than 0.05) with the traditional Chinese medicine compound 3.

Weight change, mortality and pathological changes of rabbits in each group

TABLE 2 weight change and mortality in rabbits of each group during the E.coli treatment trial

Note: the same data in the same column with the same shoulder mark letters means that the difference is not significant (P >0.05), and the difference means that the difference is significant (P < 0.05).

As can be seen from table 2, the average weight gain was significantly higher in each treatment group over the course of the trial than in the control group (P < 0.05); wherein the survival rate of the traditional Chinese medicine compound 1, diclazuril and robenidine group rabbits is 100%, and the average weight gain difference of three groups of rabbits is not significant (P is more than 0.05); the survival rate of the Chinese herbal compound 2 and the survival rate of the Chinese herbal compound 3 are 80%, the average weight gain difference between the two is not significant (P is more than 0.05), but is significantly lower than that of the Chinese herbal compound 1, the diclazuril and the robenidine group (P is less than 0.05). Beginning on day 3 after coccidial oocyst infection, rabbits developed symptoms of depression, bradykinesia, anorexia, and diarrhea. As can be seen from FIG. 2, the dead rabbits were examined by a dissecting operation to show severe congestion of the intestinal tract, duodenal dilatation, catarrhal inflammation of the intestinal mucosa, and a large amount of gas and mucosa in the small intestine.

As can be seen from the test of inhibiting the coccidian oocyst discharge, the treatment effects of the traditional Chinese medicine compound 1, the traditional Chinese medicine compound 2, the traditional Chinese medicine compound 3 and the anticoccidial drug robenidine are not obviously different 21 days after the administration; compared with another anticoccidial drug diclazuril, the traditional Chinese medicine compound 1 and the traditional Chinese medicine compound 2 have no obvious difference. The traditional Chinese medicine compound formulas 2 and 3 have poor effects on improving the survival rate of rabbits and increasing weight, and are inferior to the traditional Chinese medicine compound formula 1 in the aspects of resisting secondary infection and mucosal injury, so that the formula of the traditional Chinese medicine compound formula 1 is selected for carrying out the next treatment test.

Test for treatment

The Chinese medicine compound preparation 1 is further divided into high, medium and low 3 dosage groups to carry out treatment tests. The method comprises the steps of dividing 60 artificial infected intestinal coccidian rabbits (immunized rabbit plague seedlings) with the age of 40 days into half female rabbits and half male rabbits with the average weight of 0.8 kg-1.1 kg into 6 groups, wherein each group comprises 10 artificial infected animals, namely an infected non-traditional Chinese medicine group I, a traditional Chinese medicine high-dose group II (1g/ml), a traditional Chinese medicine medium-dose group III (0.5g/ml), a traditional Chinese medicine low-dose group IV (0.2g/ml), a diclazuril group V and a robenidine group VI. 1 x 105 of the above mixed sporulated oocysts were manually gavaged in each group of rabbits, and all the rabbits were fed with a feed without any anticoccidial drug except group VI during the test period. On days 3-9 after the oocyst inoculation, 8mL of normal saline, 8mL of 1g/mL of traditional Chinese medicine, 8mL of 0.5g/mL of traditional Chinese medicine and 8mL of 0.2g/mL of traditional Chinese medicine are respectively infused into the I-IV groups every day; group V, adding diclazuril solution into drinking water according to the concentration of 0.3 mL/L; VI group feeds with 600mg/kg robenidine feed. On day 10, the administration was stopped, and group VI was changed to a feed without added anticoccidial drugs. Recording the mental, appetite, feces changes and death conditions of the test rabbits every day, weighing the dead rabbits in time, carrying out pathological anatomy, and observing the pathological changes of each organ. The weight of each rabbit was weighed before the start and at the end of the experiment. Rabbit faeces were collected on days 7, 14 and 21 post-dose and OPG was calculated using the mcdonia ova counting method.

TABLE 3 Rabbit OPG Change after treatment

Note that the same shoulder letters in the same column of data indicate no significant difference (P >0.05), and that differences indicate significant difference (P < 0.05).

As can be seen from FIG. 3 and Table 3, after administration, the amount of oocysts excreted from rabbits in each drug group gradually decreased, and the oocysts decreased to the lowest level 14 days after administration and then gradually increased, as compared to the control group. 7 days, 14 days and 21 days after administration, the difference of the oocyst output of the drug group compared with that of the non-drug group is very obvious (P <0.01), and the difference of the oocyst output of the high-dose and medium-dose groups compared with the diclazuril group and the robenidine group is not obvious (P > 0.05); the oocyst output of the traditional Chinese medicine low-dose group after administration is higher than that of other administration groups, and the differences of the traditional Chinese medicine low-dose group and the group III and the group VI after 7 days of administration are not significant (P is more than 0.05), and the differences of the traditional Chinese medicine low-dose group and each experimental group in other stages are significant (P is less than 0.05). Compared with the traditional Chinese medicine medium-dose group III (0.5g/ml) and the traditional Chinese medicine low-dose group IV (0.2g/ml), the traditional Chinese medicine high-dose group II (1g/ml) has the least oocyst output.

Weight change and survival rate of rabbits during the test

TABLE 4 weight change and survival Rate of rabbits during the test period

Note that the same shoulder letters in the same column of data indicate no significant difference (P >0.05), and that differences indicate significant difference (P < 0.05).

As can be seen from Table 4, the survival rate of the rabbit using the medicine is obviously higher than that of the rabbit not using the medicine (P is less than 0.05) in the test period, and no rabbit dies in the high-dose group of the traditional Chinese medicine, the diclazuril group and the robenidine group. The weight gain rate of the diclazuril group, the robenidine group, the traditional Chinese medicine high-dose group, the traditional Chinese medicine low-dose group and the control group are sequentially from high to low, wherein the weight gain rate of the control group is obviously lower than that of other medicines (P is less than 0.05) except the traditional Chinese medicine low-dose group; the weight gain rate of the diclazuril group is obviously higher than that of the other drug treatment group (P < 0.05); from the aspects of weight increment and survival rate, the survival rate of the rabbits in the traditional Chinese medicine treatment group is positively correlated with the dosage of the traditional Chinese medicine.

As can be seen from treatment tests, when the concentration of the Chinese herbal compound 1 is 1g/ml, the Chinese herbal compound has better effects on reducing oocyst discharge, increasing weight and improving survival rate than the Chinese herbal compound with the concentrations of 0.5g/ml and 0.2 g/ml.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, or direct or indirect applications in other related fields, which are made by the present specification, shall be included in the scope of the present invention.

Claims (9)

1. The traditional Chinese medicine compound extract is characterized in that the raw material medicines comprise the following components in parts by weight: 40-80 parts of hawthorn, 40-80 parts of antifebrile dichroa, 40-80 parts of betel nut, 40-80 parts of liquorice, 100-150 parts of scutellaria baicalensis, 100-150 parts of coptis chinensis and 100-150 parts of Chinese herbaceous peony;

the preparation method of the extract comprises the following steps:

(1) taking raw material medicines according to parts by weight; (2) extracting with water, precipitating with ethanol, and collecting supernatant; (3) mixing the supernatants to obtain water extraction and alcohol precipitation product, and preparing the product.

2. The extract as claimed in claim 1, wherein the raw material drug comprises the following components in parts by weight: 50-70 parts of hawthorn, 50-70 parts of antifebrile dichroa, 50-70 parts of betel nut, 50-70 parts of liquorice, 110-130 parts of scutellaria baicalensis, 110-130 parts of coptis chinensis and 110-130 parts of Chinese herbaceous peony.

3. The extract as claimed in claim 2, wherein the raw material drug comprises the following components in parts by weight: 60 parts of hawthorn, 60 parts of antifebrile dichroa, 60 parts of betel nut, 60 parts of liquorice, 120 parts of scutellaria baicalensis, 120 parts of coptis chinensis and 120 parts of Chinese herbaceous peony.

4. Use of the traditional Chinese medicine compound extract as described in any one of claims 1-3 in preparation of products for treating and/or preventing rabbit intestinal coccidiosis.

5. The use according to claim 4, wherein the product further comprises pharmaceutically acceptable excipients.

6. Use according to claim 5, characterized in that the pathogen of rabbit intestinal coccidiosis is selected from Eimeria.

7. The use of claim 6, wherein the Eimeria is selected from one or more of Eimeria macroergeri, Eimeria mesogenes, Eimeria flava and Eimeria miniata.

8. Use according to claim 5, wherein the product is an inhibitor of oocyst development.

9. Use according to claim 5, wherein the product is a product for reducing the excretion of oocysts in vitro.

Images