Traditional Chinese medicine preparation for preventing and treating cryptosporidiosis and application thereof

Abstract

The invention belongs to the technical field of cryptosporidium prevention and treatment, and particularly relates to a traditional Chinese medicine preparation for preventing and treating cryptosporidiosis and application of the traditional Chinese medicine preparation. The antifebrile dichroa extract is prepared from antifebrile dichroa and used for killing cryptosporidium, and the antifebrile dichroa extract is prepared by the following steps: taking a traditional Chinese medicinal material antifebrile dichroa, and crushing the antifebrile dichroa into powder; mixing the powder with an ethanol solution with the volume fraction of 70%-80% and disodium ethylene diamine tetraacetate according to the proportion of 1kg: 8L-10L: 1g, standing for 1-4 hours, and filtering to obtain filtrate; and concentrating and drying the filtrate to obtain the antifebrile dichroa extract. The invention provides a novel scheme for preventing and treating cryptosporidium.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of Cryptosporidium prevention and treatment, and particularly relates to a Chinese medicinal preparation for preventing and treating cryptosporidiosis and application thereof. BACKGROUND

[0002] The development of Cryptosporidium can be divided into three stages, namely, schizogony, gamogony and sporogony. The oocysts of sporulation are excreted with the feces of infected hosts, and are ingested through contaminated environments, including food and drinking water. In the gastrointestinal tract or respiratory tract, the sporozoites escape from the oocysts and enter the brush border or submembrane of the epithelial cells of the respiratory tract and bursa of Fabricius, and develop into type I meronts through asexual schizogony. The type I meronts are lysed, and each meront develops into type II meronts through schizogony. The meronts lysed from the type II meronts develop into macrogametes and microgametes, and the microgametes are further divided into microgametes without flagella. The macrogametes and microgametes combine to form zygotes, and the zygotes develop into two types of oocysts, namely, thin-walled oocysts and thick-walled oocysts. After the thin-walled oocysts are broken to release sporozoites in the host, the sporozoites perform self-infection in the host. The thick-walled oocysts are excreted with the feces of the host, and can directly infect new hosts.

[0003] Based on the above development process, the harm of Cryptosporidium to animals mainly manifests in respiratory symptoms and gastrointestinal symptoms, and mainly harms young birds. For example, Cryptosporidium baileyi mainly harms chickens younger than 50 days old, and the susceptibility of chickens younger than 10 days old is the highest. The morbidity and mortality mainly occur in chickens infected at the age of 10 days or younger. Cryptosporidium also has a certain probability of infecting immunodeficient people and causing diseases such as diarrhea. Therefore, it is necessary to prevent and treat Cryptosporidium.

[0004] In the prior art, the existing methods for preventing and treating Cryptosporidium mainly include two aspects, namely, health management and drug treatment. The health management mainly refers to strengthening environmental cleaning and disinfection, especially the maintenance of public areas and sanitary facilities. The feces need to be properly treated to avoid pollution of water sources and food. The drug treatment usually refers to using antibiotics such as josamycin in animal feed, or giving infected people antibiotics such as azithromycin. However, long-term use of antibiotics is easy to make Cryptosporidium resistant, and it is necessary to continue to develop new solutions for preventing and treating Cryptosporidium. SUMMARY

[0005] In order to solve the above technical problems, the present application provides a Chinese medicinal preparation for preventing and treating cryptosporidiosis and application thereof. The Chinese medicinal preparation can effectively kill Cryptosporidium, and can be used for preventing and treating cryptosporidiosis.

[0006] The present application provides a Chinese medicinal preparation for preventing and treating cryptosporidiosis. The common perilla is made into a common perilla extract to kill Cryptosporidium. The preparation of the common perilla extract includes the following steps: Taking traditional Chinese medicinal material Changshan, grinding into powder; The powder is mixed with 70%-80% ethanol solution and disodium ethylenediaminetetraacetate according to the ratio of 1kg:8L-10L:1g, and is left for 1-4 hours, and is filtered to obtain filtrate; The filtrate is concentrated and dried to obtain Changshan extract.

[0007] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is for preventing and treating the cryptosporidium of Baishan Cryptosporidium or Turkey Cryptosporidium.

[0008] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is 80-mesh powder.

[0009] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is concentrated by rotary evaporation at 40-43 DEG C to 1 / 3-1 / 2 of the original volume.

[0010] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is dried by freeze-drying at-30 DEG C to-20 DEG C.

[0011] The application also provides a traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis, wherein the Changshan extract is mixed into basic feed to kill cryptosporidium and prevent and treat cryptosporidiosis.

[0012] The application also provides a traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis, wherein the Changshan extract is compounded with cystine and gamma-polyglutamic acid to make a feed additive; the feed additive is mixed from the following raw materials: 1000 parts of Changshan extract, 1-3 parts of cystine and 1-2 parts of gamma-polyglutamic acid.

[0013] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is mixed into feed, and the mass ratio of the feed additive to the feed is 1-1.5:1000.

[0014] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is mixed into feed, and the mass ratio of the feed additive to the feed is 1-1.5:1000.

[0015] Preferably, the traditional Chinese medicinal preparation for preventing and treating cryptosporidiosis is mixed into feed, and the mass ratio of the feed additive to the feed is 1-1.5:1000.

[0016] Compared with the prior art, the application has the following beneficial effects: The application adopts a special extraction process to prepare the common perilla extract. In the extraction process, in the presence of disodium ethylenediaminetetraacetate, the alkaloid toxin in the common perilla forms a selective dissolution relationship with common perilla ketone. That is, after adjusting the ion environment of the extraction solution by disodium ethylenediaminetetraacetate, the alkaloid is more adsorbed by the common perilla medicinal material and is not easy to dissolve in the 70%-80% ethanol solution, while the common perilla ketone is easy to dissolve in the 70%-80% ethanol solution. According to the experimental results of the application, the content of common perilla ketone in the common perilla extract is more than 63.29 mg / kg, and the total content of alkaloids is less than 2.05 mg / g.

[0017] In order to kill cryptosporidium and improve the effect of preventing and treating cryptosporidiosis, the application takes the traditional Chinese medicine common perilla as the main raw material, extracts active ingredients therefrom to obtain a common perilla extract, which can reduce the toxicity of the original common perilla and reduce side effects.

[0018] In other schemes provided by the application, the common perilla extract is compounded with cystine and gamma-polyglutamic acid to form a feed additive after mixing, so as to prevent and treat cryptosporidium and also produce good effects. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 The technical roadmap of the application. DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to better understand the technical solutions of the application and implement them, the application will be further described below in combination with specific embodiments.

[0021] In the description of the application, unless otherwise specified, the reagents used are commercially available, and the methods used are conventional techniques in the art.

[0022] The technical roadmap of the application is shown in Figure 1 The inventive concept of the application is as follows:

[0023] The common perilla ketone is rich in the common perilla and is one of the active components of the common perilla, can block the growth and reproduction of parasites, slow down the reproduction speed of cryptosporidium, and ultimately cause the death of the cryptosporidium, thereby playing a pesticidal effect. According to the analysis of the theory of traditional Chinese medicine, the common perilla is bitter and cold, and belongs to the lung, liver and spleen channels, and has the effects of clearing heat and resolving toxins and killing and expelling parasites, thereby relieving the symptoms of diarrhea, damp-heat diarrhea and other infections caused by cryptosporidium. The existing technology usually extracts the common perilla with a 70%-80% ethanol solution, dries to obtain an extract, and then uses it to prevent and kill or eliminate cryptosporidium, but the content of the common perilla ketone is not high, and the prevention effect is also poor.

[0024] The application provides a new scheme for preventing and treating Cryptosporidium, and relates to a traditional Chinese medicine preparation for preventing and treating cryptosporidiosis, for example, common perilla prepared into common perilla extract for killing Cryptosporidium.

[0025] In order to improve the content of common perilla ketone in the common perilla extract, the application provides the following preparation method. The common perilla is taken as a traditional Chinese medicine material, and the common perilla below is the dried root of common perilla, and will not be repeated, and the application does not involve the research on different medicinal parts, and is crushed into powder; The powder is mixed with 70-80% ethanol solution and disodium ethylenediaminetetraacetate at a ratio of 1 kg: 8-10 L: 1 g, and is placed for 1-4 hours, and is filtered to obtain a filtrate; The filtrate is concentrated and dried to obtain the common perilla extract.

[0026] In the above extraction method, under the condition that disodium ethylenediaminetetraacetate exists, the alkaloid toxin in common perilla forms a selective dissolution relationship with common perilla ketone, that is, after the ion environment in the extraction solution is adjusted by disodium ethylenediaminetetraacetate, the adsorption of the alkaloid and the common perilla medicinal material is stronger, and the alkaloid is not easy to dissolve in the 70-80% ethanol solution, while the common perilla ketone is easy to dissolve in the 70-80% ethanol solution.

[0027] Preferably, the powder is 80-mesh powder, which can increase the contact area of the medicine and the extraction reagent. The concentration condition is that the filtrate is concentrated to 1 / 3-1 / 2 of the original volume by rotary evaporation at 40-43 DEG C, the evaporation temperature is relatively mixed, and the evaporation speed can also be considered. The drying condition is-30 DEG C to-20 DEG C freeze drying, which is a relatively easy drying condition.

[0028] According to the experimental results of the application, the content of common perilla ketone in the common perilla extract is more than 63.29 mg / kg, and the total content of alkaloids is less than 2.05 mg / g.

[0029] The application takes common perilla as a traditional Chinese medicine as a main raw material, extracts active ingredients, obtains common perilla extract, can reduce the toxicity of the original common perilla, and reduces side effects.

[0030] The application also mixes the common perilla extract with cystine (L-cystine purchased from Jiangsu Jiujia Biological Technology Co., Ltd., food grade) and gamma-polyglutamic acid (Guangdong Mingcheng Biological Technology Co., Ltd., CAS No. 25513-46-6, molecular weight 147.13, product grade agricultural grade) to form a feed additive for preventing and treating Cryptosporidium. The feed additive is mixed by the following raw materials in weight parts: common perilla extract 1000 parts, cystine 1-3 parts, and gamma-polyglutamic acid 1-2 parts.

[0031] The above-mentioned extract of common perilla or feed additive has a good prevention and treatment effect on Cryptosporidium baileyi or Cryptosporidium meleagridis.

[0032] The following are some embodiments and controls of the present application. The raw materials of the following embodiments and controls are derived from the same as in Example 1, and the differences are indicated by different embodiment and control contents.

[0033] Example 1 A Chinese medicine preparation for preventing and treating cryptosporidiosis is prepared according to the following method: Take Chinese herbal medicine common perilla and crush it into a powder with a particle size of 80 mesh; Mix the powder with a 70% ethanol solution and disodium ethylenediaminetetraacetate according to a ratio of 1 kg: 8 L: 1 g, stand for 1 h, filter, and obtain a filtrate; Concentrate the filtrate to 1 / 3 of the original volume by rotary evaporation at 40℃, and obtain a concentrated solution; Freeze-dry the concentrated solution at -20℃, and obtain an extract of common perilla.

[0034] Example 2 A Chinese medicine preparation for preventing and treating cryptosporidiosis is prepared according to the following method: Take Chinese herbal medicine common perilla and crush it into a powder with a particle size of 80 mesh; Mix the powder with a 80% ethanol solution and disodium ethylenediaminetetraacetate according to a ratio of 1 kg: 8 L: 1 g, stand for 1 h, filter, and obtain a filtrate; Concentrate the filtrate to 1 / 3 of the original volume by rotary evaporation at 40℃, and obtain a concentrated solution; Freeze-dry the concentrated solution at -20℃, and obtain an extract of common perilla.

[0035] Example 3 A Chinese medicine preparation for preventing and treating cryptosporidiosis is prepared according to the following method: Take Chinese herbal medicine common perilla and crush it into a powder with a particle size of 80 mesh; Mix the powder with a 70% ethanol solution and disodium ethylenediaminetetraacetate according to a ratio of 1 kg: 10 L: 1 g, stand for 1 h, filter, and obtain a filtrate; Concentrate the filtrate to 1 / 3 of the original volume by rotary evaporation at 40℃, and obtain a concentrated solution; Freeze-dry the concentrated solution at -20℃, and obtain an extract of common perilla.

[0036] Example 4 A Chinese medicine preparation for preventing and treating cryptosporidiosis is prepared according to the following method: Take Chinese herbal medicine common perilla and crush it into a powder with a particle size of 80 mesh; The powder is mixed with 70% ethanol solution and disodium ethylenediaminetetraacetate at a ratio of 1 kg:8 L:1 g, left to stand for 4 h, filtered to obtain a filtrate; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0037] Example 5 A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is crushed into powder of 80 mesh; The powder is mixed with 70% ethanol solution and disodium ethylenediaminetetraacetate at a ratio of 1 kg:8 L:1 g, left to stand for 1 h, filtered to obtain a filtrate; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 2 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0038] Example 6 A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is crushed into powder of 80 mesh; The powder is mixed with 70% ethanol solution and disodium ethylenediaminetetraacetate at a ratio of 1 kg:8 L:1 g, left to stand for 1 h, filtered to obtain a filtrate; The filtrate is concentrated by rotary evaporation at 43°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0039] Example 7 A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is crushed into powder of 80 mesh; The powder is mixed with 70% ethanol solution and disodium ethylenediaminetetraacetate at a ratio of 1 kg:8 L:1 g, left to stand for 1 h, filtered to obtain a filtrate; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -35°C to obtain the Radix Dichondrae extract.

[0040] Control group 1 (without disodium ethylenediaminetetraacetate) A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is crushed into powder of 80 mesh; The powder is mixed with a 70% ethanol solution in a ratio of 1 kg:8 L, left to stand for 1 h, filtered, and a filtrate is obtained; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0041] Control group 2 (ethylenediaminetetraacetic acid disodium salt is used in too low an amount) A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is pulverized into a powder of 80 mesh; The powder is mixed with a 70% ethanol solution and ethylenediaminetetraacetic acid disodium salt in a ratio of 1 kg:8 L:0.5 g, left to stand for 1 h, filtered, and a filtrate is obtained; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0042] Control group 3 (ethylenediaminetetraacetic acid disodium salt is used in too high an amount) A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is pulverized into a powder of 80 mesh; The powder is mixed with a 70% ethanol solution and ethylenediaminetetraacetic acid disodium salt in a ratio of 1 kg:8 L:2 g, left to stand for 1 h, filtered, and a filtrate is obtained; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0043] Control group 4 (replaced with the analogue ethylenediaminetetraacetic acid) A Chinese medicinal preparation for preventing and treating cryptosporidiosis is prepared according to the following method: The Chinese medicinal material Radix Dichondrae is pulverized into a powder of 80 mesh; The powder is mixed with a 70% ethanol solution and ethylenediaminetetraacetic acid in a ratio of 1 kg:8 L:1 g, left to stand for 1 h, filtered, and a filtrate is obtained; The filtrate is concentrated by rotary evaporation at 40°C to 1 / 3 of the original volume to obtain a concentrated solution; The concentrated solution is freeze-dried at -20°C to obtain the Radix Dichondrae extract.

[0044] To evaluate the quality and safety of the common perilla extract, the content of the traditional Chinese medicine insecticidal component of common perilla ketone and the content of the alkaloid that is easy to cause toxicity in the common perilla extract of different embodiments and control groups are tested, and the results are shown in Table 1. The results show that, after the addition of disodium ethylenediaminetetraacetate for adjustment, the content of the alkaloid is reduced, and the content of the common perilla ketone is increased. The alkaloid toxin in the common perilla and the common perilla ketone form a selective dissolution relationship. In addition, the results also show that, even if the similar substance ethylenediaminetetraacetic acid is used, due to the lack of the effect of sodium element, the content of the common perilla ketone is still not high.

[0045] Table 1 Total content of common perilla ketone and alkaloid To verify the insecticidal effect of the common perilla extract, the cryptosporidium baileyi and the cryptosporidium meleagridis are selected for experiments. The physiological characteristics of the cryptosporidium baileyi are as follows: the oocysts are elliptical or oval, mainly parasitize in the respiratory tract, bursa of fabricius and intestinal mucosa epithelial cells of poultry, and cause respiratory diseases and diarrhea. The physiological characteristics of the cryptosporidium meleagridis are as follows: the oocysts are small and approximately circular, mainly parasitize in the intestinal tract of poultry, and cause diarrhea. The extracts of the embodiment 1 to the embodiment 7 and the control group 1 to the control group 4 are mixed with the cryptosporidium baileyi or the cryptosporidium meleagridis respectively, and the inactivation rate of the two cryptosporidium is detected.

[0046] The operation method of the cryptosporidium baileyi experiment is as follows: 100 mL of the worm sample (cryptosporidium baileyi) with a concentration of 2×10 4 individuals / mL is placed in a container, the temperature of the environment where the container is located is controlled to be 25℃ (room temperature), the pH of the worm sample is adjusted to be 7.0, 1 g of the common perilla extract is added, and the action time is 25 min. The fluorescence vital staining method (reference literature: Li Shao-feng, Chu Hong-ping, Ran Zhi-lin, et al. Influence factors and mechanism of ultraviolet / ultrasonic synergistic inactivation of cryptosporidium [J]. Environmental Science Research, 2017, 30(8): 6. DOI:10.13198 / j.issn.1001-6929.2017.02.55.) is used to detect and calculate the inactivation rate. The worm sample without the common perilla extract is taken as a blank control.

[0047] The inactivation rate experiment and calculation method of the cryptosporidium meleagridis are the same as those of the cryptosporidium baileyi, and only the cryptosporidium species is changed to the cryptosporidium meleagridis.

[0048] The inactivation rate detection results of the two cryptosporidium are shown in Table 2. The results of Table 2 show that the schemes of the embodiment 1 to the embodiment 7 have good anti-worm effects on the two cryptosporidium. The anti-worm effects of the control group 1 to the control group 5 are all not ideal.

[0049] Table 2 Anti-worm experiment results of different common perilla extracts Based on the same inventive concept, the present application also provides a feed additive. The following examples are included.

[0050] Example 8 A feed additive is prepared by compounding the extract of Brucea javanica in Example 1 with cystine and gamma-polyglutamic acid; the feed additive is mixed from the following raw materials: 1000 parts of the extract of Brucea javanica, 1 part of cystine, and 1 part of gamma-polyglutamic acid.

[0051] The extract of Brucea javanica is prepared according to the following method: Take the Chinese herbal medicine Brucea javanica and crush it into a powder with a mesh size of 80; Mix the powder with a 70% ethanol solution and disodium ethylenediaminetetraacetate according to a ratio of 1 kg:8 L:1 g, let it stand for 1 h, filter, and obtain the filtrate; Concentrate the filtrate to 1 / 3 of the original volume by rotary evaporation at 40℃, and obtain the concentrated solution; Freeze-dry the concentrated solution at -20℃, and obtain the extract of Brucea javanica.

[0052] Example 9 A feed additive is prepared by compounding the extract of Brucea javanica in Example 1 with cystine and gamma-polyglutamic acid; the feed additive is mixed from the following raw materials: 1000 parts of the extract of Brucea javanica, 3 parts of cystine, and 1 part of gamma-polyglutamic acid.

[0053] The extract of Brucea javanica is prepared according to the following method: Take the Chinese herbal medicine Brucea javanica and crush it into a powder with a mesh size of 80; Mix the powder with a 70% ethanol solution and disodium ethylenediaminetetraacetate according to a ratio of 1 kg:8 L:1 g, let it stand for 1 h, filter, and obtain the filtrate; Concentrate the filtrate to 1 / 3 of the original volume by rotary evaporation at 40℃, and obtain the concentrated solution; Freeze-dry the concentrated solution at -20℃, and obtain the extract of Brucea javanica.

[0054] Example 10 A feed additive is prepared by compounding the extract of Brucea javanica in Example 1 with cystine and gamma-polyglutamic acid; the feed additive is mixed from the following raw materials: 1000 parts of the extract of Brucea javanica, 1 part of cystine, and 2 parts of gamma-polyglutamic acid.

[0055] The extract of Brucea javanica is prepared according to the following method: Take the Chinese herbal medicine Brucea javanica and crush it into a powder with a mesh size of 80; Mix the powder with a 70% ethanol solution and disodium EDTA at a ratio of 1 kg: 8 L: 1 g, let stand for 1 h, filter, and obtain the filtrate. The filtrate was concentrated to 1 / 3 of its original volume by rotary evaporation at 40°C to obtain a concentrated solution. The concentrate was freeze-dried at -20°C to obtain Changshan extract.

[0056] Control group 5 One type of feed additive is cystine.

[0057] Control group 6 One type of feed additive is γ-polyglutamic acid.

[0058] Control group 7 A feed additive is the Changshan extract from Example 1. The experimental results of this control group correspond to the results of Example 1.

[0059] The Changshan extract was prepared according to the following method: Take the Chinese medicinal herb Changshan and grind it into 80-mesh powder; Mix the powder with a 70% ethanol solution and disodium EDTA at a ratio of 1 kg: 8 L: 1 g, let stand for 1 h, filter, and obtain the filtrate. The filtrate was concentrated to 1 / 3 of its original volume by rotary evaporation at 40°C to obtain a concentrated solution. The concentrate was freeze-dried at -20°C to obtain Changshan extract.

[0060] Insect resistance experiments and feeding experiments were conducted using the above-mentioned feed additives, and the specific procedures are as follows.

[0061] (1) Insect resistance experiment, the experimental method is similar to that described above, as follows: The experimental procedure for Cryptosporidium benziii is as follows: Prepare 100 mL of a solution with a concentration of 2×10⁻⁶... 4 The number of insect samples (Cryptospora benziae) per mL was placed in a container, and the ambient temperature of the container was controlled at 25℃ (room temperature). The pH of the insect samples was adjusted to 7.0, and 1g of feed additive was added. After 25 min, the inactivation rate was detected and calculated by fluorescent live staining (Reference: Li Shaofeng, Chu Hongping, Ran Zhilin, et al. Influencing factors and mechanisms of UV / ultrasound synergistic inactivation of Cryptosporidium [J]. Environmental Science Research, 2017, 30(8):6.DOI:10.13198 / j.issn.1001-6929.2017.02.55.). Insect samples without Changshan extract were used as blank controls.

[0062] The inactivation rate experiment and calculation method for Cryptosporidium turkeyis are the same as those for Cryptosporidium benziii, except that Cryptosporidium is replaced with Cryptosporidium turkeyis, and the added substance is also a feed additive.

[0063] The results are shown in Table 3. The results in Table 3 show that the feed additive of Example 8 to Example 10 has good anti-parasitic effect on the two Cryptosporidium. Compared with Examples, the anti-Cryptosporidium effect of cystine and γ-polyglutamic acid alone in Control Group 5 and Control Group 6 is weak, but the anti-parasitic effect is significantly enhanced after compounding with common perilla extract.

[0064] Table 3 Anti-parasitic experimental results of different feed additives (2) Feeding experiment The feed additive is mixed into the basic feed, and the mass ratio of the feed additive to the basic feed is 1:1000. The basic feed is chicken feed without anti-Cryptosporidium drugs, and the basic feed is mixed by the following raw materials in percentage by weight: soybean meal 28%, fish meal 2%, soybean oil 1%, calcium bicarbonate 1%, experiment 0.3%, and corn to make up 100%, noting that the particle size of each raw material needs to be less than 1 mm.

[0065] The concentration of the Cryptosporidium parvum oocyst sample is configured to be 1 x 10 5

[0066] Grouping: 210 healthy 3-day-old Qingyuan chickens were selected, each chicken weighing 45±5g, and randomly divided into 7 groups, 30 chickens in each group, half male and half female.

[0067] Treatment: The blank control group is only fed with the basic feed.

[0068] Example 8 group, after each chicken is gavaged with 1 mL of Cryptosporidium parvum oocyst sample, the common perilla extract of Example 8 is mixed into the basic feed for feeding, and the mass ratio of the common perilla extract to the basic feed is 1:1000.

[0069] Control Group 5 experimental group, after each chicken is gavaged with 1 mL of Cryptosporidium parvum oocyst sample, the common perilla extract of Control Group 5 is mixed into the basic feed for feeding, and the mass ratio of the common perilla extract to the basic feed is 1:1000.

[0070] Control Group 6 experimental group, after each chicken is gavaged with 1 mL of Cryptosporidium parvum oocyst sample, the common perilla extract of Control Group 6 is mixed into the basic feed for feeding, and the mass ratio of the common perilla extract to the basic feed is 1:1000.

[0071] Control Group 7 experimental group, after each chicken is gavaged with 1 mL of Cryptosporidium parvum oocyst sample, the common perilla extract of Control Group 7 is mixed into the basic feed for feeding, and the mass ratio of the common perilla extract to the basic feed is 1:1000.

[0072] ​The positive control group, each chicken was given 1 mL of Besnoeia besnoiti oocysts, and no feed additive or common mountain extract was fed.

[0073] The drug control group, each chicken was given 1 mL of Besnoeia besnoiti oocysts, and the paromomycin was mixed into the basic feed, and the mass ratio of paromomycin to basic feed was 1:1000.

[0074] The amount of water and feed in each group was the same.

[0075] The efficacy determination method mainly depends on the reduction rate of oocysts after infection; the feces is collected daily from the third day after infection, and the monitoring is continuously conducted for 15 days. The purified cysts are collected, the oocyst morphology is observed under a microscope, and the counting is conducted.

[0076] Symptom observation: The positive control group inoculated with Besnoeia besnoiti oocysts showed depression, rough and messy feathers on the third day, and obvious diarrhea symptoms from the fifth day, and no chick died during the period, which proved that the disease model was successfully constructed. The blank control group was healthy chicks, and no diarrhea symptoms and death were observed.

[0077] The average number of oocysts per gram of feces (i.e. OPG value) and the oocyst reduction rate of each group were calculated, and the results are shown in Table 4. The groups of example 8 and the control group 7 (common mountain extract of example 1) have good anti-parasite effect on Besnoeia besnoiti.

[0078] Table 4 Anti-parasite experimental results of different feed additives Note: “-” indicates that this item is not calculated.

[0079] It should be noted that when the numerical range is involved in the present application, it should be understood that each numerical range of the two endpoints and any number between the two endpoints can be selected. Since the same steps and examples are used, in order to prevent repetition, the present application describes the preferred examples. Although the preferred embodiments of the present application have been described, those skilled in the art can make further changes and modifications to these embodiments once they know the inventive concept of the present application, and these changes and modifications all fall within the scope of the present application.

[0080] Obviously, those skilled in the art can make various modifications and variations to the present application without departing from the spirit and scope of the present application. If these modifications and variations of the present application fall within the scope of the equivalent technology of the present application, the present application also intends to include these modifications and variations.

Claims

1. A traditional Chinese medicine preparation for the prevention and treatment of cryptosporidiosis, characterized in that, The common perilla is made into common perilla extract after killing cryptosporidium, and the preparation of the common perilla extract comprises the following steps: Taking Chinese herbal medicine common perilla, crushing into powder; Mixing the powder with 70%-80% ethanol solution and disodium ethylenediaminetetraacetate according to the ratio of 1kg:8L-10L:1g, standing for 1h-4h, filtering to obtain filtrate; Concentrating and drying the filtrate to obtain common perilla extract.

2. The Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, The cryptosporidium is Cryptosporidium baileyi or Cryptosporidium meleagridis.

3. The Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, The powder is 80 mesh powder.

4. The Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, The concentration condition is that the filtrate is concentrated to 1 / 3-1 / 2 of the original volume by rotary evaporation at 40-43 DEG C.

5. The Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, The drying condition is freeze drying at-30 DEG C to-20 DEG C.

6. The use of the Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, Mixing the common perilla extract into basal feed.

7. The use of the Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 1, characterized in that, Compounding the common perilla extract with cystine and gamma-polyglutamic acid to make feed additive, and mixing into basal feed; The feed additive is mixed by the following raw materials: common perilla extract 1000 parts, cystine 1-3 parts, gamma-polyglutamic acid 1-2 parts.

8. The use of the Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 7, characterized in that, The mass ratio of feed additive to feed is 1-1.5:1000.

9. The use of the Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 8, characterized in that, The basal feed is poultry feed.

10. The Chinese medicine preparation for preventing and treating cryptosporidiosis according to claim 9, characterized in that, The poultry is chicken.

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