CN107349376B - Anti-coccidiosis probiotic medicine and preparation method and application thereof - Google Patents

Anti-coccidiosis probiotic medicine and preparation method and application thereof Download PDF

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CN107349376B
CN107349376B CN201710517764.1A CN201710517764A CN107349376B CN 107349376 B CN107349376 B CN 107349376B CN 201710517764 A CN201710517764 A CN 201710517764A CN 107349376 B CN107349376 B CN 107349376B
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traditional chinese
probiotic
coccidiosis
chinese medicine
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CN107349376A (en
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王玲
郭志廷
杨峰
张彬
郭文柱
魏小娟
罗金印
周绪正
牛建荣
杨珍
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Lanzhou Institute of Animal Husbandry and Veterinary Medicine CAAS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/11—Pteridophyta or Filicophyta (ferns)
    • A61K36/12—Filicopsida or Pteridopsida
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/47—Euphorbiaceae (Spurge family), e.g. Ricinus (castorbean)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/484—Glycyrrhiza (licorice)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/75—Rutaceae (Rue family)
    • A61K36/752—Citrus, e.g. lime, orange or lemon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9064—Amomum, e.g. round cardamom
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Abstract

The invention provides a preparation method of a chicken coccidiosis resistant probiotic drug, wherein the probiotic drug is prepared by carrying out facultative anaerobic fermentation on a traditional Chinese medicine compound extracting solution by saccharomyces cerevisiae; the traditional Chinese medicine compound extracting solution is prepared by mixing the following raw materials in parts by mass and then carrying out water extraction: 60-90 parts of antifebrile dichroa, 30-60 parts of tsaoko amomum fruits, 40-60 parts of humifuse euphorbia herb, 30-50 parts of purslane, 20-40 parts of copperleaf herb, 10-30 parts of Chinese silkvine, 10-30 parts of liquorice, 20-40 parts of dark plum, 20-50 parts of dried orange peel and 10-30 parts of medicated leaven. The invention also provides the anti-coccidiosis probiotic medicine prepared by the method and application thereof. The probiotic medicament has a good treatment effect on chicken coccidiosis.

Description

Anti-coccidiosis probiotic medicine and preparation method and application thereof
Technical Field
The invention belongs to the technical field of veterinary drugs and microecological drugs, and particularly relates to a chicken coccidiosis resistant probiotic drug, and a preparation method and application thereof.
Background
Chicken coccidiosis is an intestinal parasitic disease with epidemic, high morbidity and high mortality in global distribution, and the poultry mortality of 5-10% is caused by the intestinal parasitic disease, which can cause the growth of poultry to be blocked and the feed to be transferredThe reduction of the digestion rate and the reduction of the quality of meat and eggs. Eimeria tenella (E.tenella) among all coccidioides of the genus CoccidiumE.tenella) The harm to the chickens is the most serious, and the harm is mainly caused by chicken cecal epithelial cells. The global economic loss caused by the disease is about 50 billion dollars each year, and the national cost is about 30 billion yuan, wherein the cost of the anticoccidial drug is about 6 billion yuan. At present, the chemical synthetic drugs are mainly used at home and abroad to prevent and treat the coccidiosis of the chicken, such as diclazuril, toltrazuril, halofuginone and the like, which greatly contribute to the healthy development of the modern chicken industry, and more research is made on the coccidiosis resisting effect and mechanism at home and abroad. With the increasing development of coccidium drug resistance, the lack of general application of immunoprophylaxis, the serious problem of drug residue in anticoccidial chemical drugs and the continuous enhancement of health consciousness of people, the research and development of safe, environment-friendly, broad-spectrum and efficient anticoccidial drugs are urgently needed.
Therefore, the poultry industry is more inclined to use the medicines with strong pertinence, high bioavailability and strong safety in the susceptible day age of the chicks, and the medicines are prevented and treated through a convenient oral way, so that the defects of the existing medicine condition are overcome.
The traditional Chinese medicine dichroa febrifuga has the effects of killing insects, intercepting malaria and eliminating phlegm, can be used independently or used as a main drug of a compound preparation as a traditional Chinese medicine with anticoccidial activity, and effective active monomers dichroine (dichroine B, beta-dichroine) and dichroine A (isobificine, dichroine A, alpha-dichroine) are anticoccidial effective components in dichroa febrifuga extract. The research shows that the toxicity and the curative effect of dichroa febrifuga B are higher than those of dichroa febrifuga A, and compared with dichroa febrifuga A, the dichroa febrifuga B has stronger antimalarial effect which is about 100 times that of dichroa febrifuga A. In view of the fact that dichroa febrifuga is a traditional Chinese medicinal material with small toxicity, the extract has the effect of promoting vomiting in clinical medication, and the content of the effective active ingredient dichroa febrifuga total alkaloid in the traditional Chinese medicine compound for resisting coccidiosis is low at present, thereby bringing uncertain factors to clinical safe use. In addition, the extraction rate of effective active ingredients is low due to the difficulty in extracting and separating the dichroa febrifuga ethyl element and the dichroa febrifuga methyl element in the dichroa febrifuga original medicinal materials or decoction pieces.
Disclosure of Invention
The invention provides a chicken coccidiosis-resistant probiotic medicament and a preparation method and application thereof, and the probiotic medicament provided by the invention has the effects of killing coccidiosis, cooling blood and stopping dysentery, clearing heat and drying dampness, and strengthening spleen and regulating qi, and is suitable for treating chicken coccidiosis and mixed infection of coccidiosis and bacteria.
The invention provides a preparation method of a chicken coccidiosis resistant probiotic drug, wherein the probiotic drug is prepared by carrying out facultative anaerobic fermentation on a traditional Chinese medicine compound extracting solution by saccharomyces cerevisiae; the traditional Chinese medicine compound extracting solution is prepared by mixing the following raw materials in parts by mass and then carrying out water extraction: 60-90 parts of antifebrile dichroa, 30-60 parts of tsaoko amomum fruits, 40-60 parts of humifuse euphorbia herb, 30-50 parts of purslane, 20-40 parts of copperleaf herb, 10-30 parts of Chinese silkvine, 10-30 parts of liquorice, 20-40 parts of dark plum, 20-50 parts of dried orange peel and 10-30 parts of medicated leaven.
Preferably, the traditional Chinese medicine compound extracting solution is prepared by mixing the following raw material medicines in parts by mass and then carrying out water extraction: 80 parts of antifebrile dichroa, 40 parts of tsaoko amomum fruits, 50 parts of humifuse euphorbia herbs, 40 parts of purslane, 30 parts of copperleaf herbs, 20 parts of Chinese silkvine, 20 parts of liquorice, 30 parts of dark plums, 40 parts of dried orange peels and 20 parts of medicated leavens.
Preferably, the preparation method of the traditional Chinese medicine compound extracting solution comprises the following steps: crushing the raw materials, weighing the raw materials according to the formula, adding deionized water with the weight 6-10 times of the total weight of the traditional Chinese medicines, soaking for 30-60 minutes, decocting for 40-60 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, filtering to remove residues, and mixing the above 3 times of decoction filtrate.
Preferably, the fermentation method of the traditional Chinese medicine compound extracting solution comprises the following steps: 200 g/L maltose and 200mL/L, MgSO malt extract are added into the traditional Chinese medicine compound extracting solution4 5.0 g/L、FeSO4 3.0g/L, sterilizing, cooling to 28-32 deg.C, inoculating Saccharomyces cerevisiae with an inoculum size of 8-10%, performing facultative anaerobic fermentation culture at 35 deg.C in an aseptic fermentation chamber with shaking table at a speed of 40r/min until white mycoderm grows on the liquid surface.
Preferably, the strain number of the saccharomyces cerevisiae is CVCC 2.1011, and the strain content is 9.0 multiplied by 108 CFU/mL。
The invention also provides the anti-coccidiosis probiotic medicine prepared by the method.
Preferably, the liquid surface mycoderm of the fermentation filtrate obtained by fermentation is removed to obtain the oral drinking water agent of the probiotic medicine.
Preferably, the fermentation filtrate obtained by fermentation is centrifugally concentrated, preferably, the centrifugal concentration is carried out at the temperature of 4 ℃, the rotation speed of 5000-7000rpm and the centrifugal time of 8-10min, the precipitate is collected, a basic freeze-drying protective agent and a protein stabilizing agent are added into the precipitate, the mixture is uniformly mixed, the pH value is adjusted, the balance is carried out, and the powder of the probiotic medicament is obtained after freeze drying.
The invention also provides application of the anti-chicken coccidiosis probiotic medicine in preparation of a medicine or feed additive for treating chicken coccidiosis.
Preferably, the chicken coccidiosis is chicken coccidiosis caused by Eimeria tenella.
The traditional Chinese medicine compound is prepared according to the principles of clearing heat, preventing malaria, eliminating phlegm, drying dampness, cooling blood, stopping dysentery, invigorating spleen and regulating qi according to certain weight parts according to the theory of Chinese beast medicine and modern traditional Chinese medicine, and comprises dichroa root, tsaoko amomum fruit, humifuse euphorbia herb, purslane, acalypha australis, Chinese silkvine root, liquorice, dark plum, dried orange peel and medicated leaven. The formula principle is as follows: according to the theory of syndrome differentiation and treatment by Chinese veterinarian, chicken coccidiosis is exogenous damp-heat, the damp-heat is accumulated in large intestine, and heat toxin is forced to flow recklessly to cause feces with blood and dampness to stagnate in spleen soil, so that chicken diarrhea is caused. In the formula, the dichroa febrifuga and the amomum tsao-ko are used together to play the functions of clearing heat, promoting the secretion of saliva or body fluid and resisting malaria, and the amomum tsao-ko can strengthen the antimalarial efficacy of the dichroa febrifuga and avoid the defect of vomiting from the dichroa febrifuga; the humifuse euphorbia herb, the purslane and the copperleaf herb are beneficial to clearing away damp-heat and astringing to stop bleeding; the black chive is helpful for detoxification; the dark plum, the dried orange peel and the medicated leaven are matched to facilitate the production of body fluid and the diarrhea, and the spleen strengthening and the qi benefiting; the licorice has the functions of purging fire, detoxicating, relieving pain, invigorating spleen and replenishing qi, and harmonizing the effects of the other medicines. The whole formula is precise in compatibility, and the traditional Chinese medicine compound extract prepared by the formula has the effects of resisting malaria, eliminating phlegm, clearing heat, stopping diarrhea, cooling blood, stopping dysentery, regulating qi and eliminating dampness.
After the active yeast culture and the facultative anaerobic fermentation are carried out on the traditional Chinese medicine compound extracting solution, the conversion and the decomposition of polysaccharide, starch, cellulose and other pharmaceutical active ingredients in the traditional Chinese medicine compound are facilitated, more effective pharmaceutical active ingredients are obtained, the toxic and side effects of the medicine are reduced, and the palatability of the medicine is improved.
The active yeast culture has the effects of anaerobically fermenting the traditional Chinese medicine extracting solution and improving the content and the utilization rate of the effective active ingredients of the traditional Chinese medicine, and the nutrient ingredients of the active yeast culture can stimulate the defense capacity of the immune system of the chicks, reduce the generation of toxic substances in intestines, have the promotion effect on the immune function of organisms, enhance the immunity of the chicks, improve the cure rate, have no drug resistance and effectively control the occurrence of coccidiosis of the chicks.
The probiotic pharmaceutical composition for preventing and treating chicken coccidiosis provided by the invention is prepared by anaerobic fermentation of an active yeast culture traditional Chinese medicine compound extract, has higher effective active ingredients and utilization rate of the medicine, and has the effects of killing coccidium, clearing heat, drying dampness, cooling blood, stopping dysentery, strengthening spleen, tonifying qi, increasing the number of beneficial flora in intestinal tract, improving disease resistance and immune function of chicks and the like. The preparation has the advantages of convenient material selection, high extraction efficiency, relatively low cost, easy formation of large-scale production and capability of being prepared into oral administration preparations convenient for veterinary clinical application.
The invention prepares the anticoccidial traditional Chinese medicine compound medicine according to the principles of clearing heat, checking malaria, eliminating phlegm, drying dampness, cooling blood, stopping dysentery, strengthening spleen and regulating qi according to the theories of Chinese veterinary medicine and modern traditional Chinese medicine and pharmacology. By applying the modern Chinese medicine extraction and separation technology, the effective active ingredients of the Chinese medicine compound are extracted, and the probiotic fermentation technology is combined to decompose and convert the effective pharmaceutical ingredients such as alkaloid, polysaccharide, starch, cellulose and the like in the compound extracting solution, so that a more effective active monomer is obtained, the toxic and side effects of antifebrile dichroa are reduced, the palatability of the medicine is improved, and the effective active pharmaceutical ingredients and the utilization rate of the medicine are further improved. Meanwhile, the nutrient components of the probiotics can stimulate the defense capacity of the immune system of the chicken, reduce the generation of toxic substances in the intestines, have the effect of promoting the immune function of the organism, enhance the immunity of the chicks, improve the cure rate, have no drug resistance and effectively control the generation of coccidiosis in the chicken.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples are commercially available unless otherwise specified. The Chinese medicinal raw materials used in the invention can be purchased from pharmaceutical shops or reagent companies, and the specification of the Chinese medicinal raw materials meets the national or industrial standard.
Example 1 preparation of active yeast culture, the procedure was as follows:
(1) preparation of a culture medium:
liquid culture medium: the components are as follows: 50 g/L glucose, 2.0 g/L yeast extract and 120 mL/L, MgSO malt extract40.5 g/L、FeSO40.1 g/L and 1.0 g/L of urea. Adjusting pH to 6.0, sterilizing at 121 deg.C for 20 min, and packaging into conical flask.
Slant and plate media: the components are as follows: 50 g/L glucose, 2.0 g/L yeast extract and 120 mL/L, MgSO malt extract4 0.5 g/L、FeSO40.1 g/L, 1.0 g/L urea and 20 g/L agar. Adjusting pH to 6.0, sterilizing at 121 deg.C for 20 min, and packaging into conical flask.
(2) Culturing yeast:
saccharomyces cerevisiae (Saccharomyces cerevisiaeThe culture is purchased from China veterinary microbial strain preservation and management center and is a feed yeast, and the strain number is CVCC 2.1011), a ring of yeast strains with rich growth is selected from slant strain storage, the yeast strains are inoculated in 50 mL of liquid culture medium and cultured at the constant temperature of 30 ℃ for about 24 hours, the logarithmic phase can be reached, the bacterial liquid is shaken evenly, the bacterial liquid is inoculated in 100 mL of liquid culture medium, and the culture is carried out for 24 hours under the same condition, so that the rejuvenating yeast is obtained.
Transferring and expanding culture: inoculating the rejuvenation yeast into a liquid culture medium, performing shake culture at 30 ℃ and 40r/min, and observing. The inoculum size of the strain expanded subculture is controlled at 6-10%, the liquid loading amount is less than 60% to ensure the dissolved oxygen amount, the culture temperature is 30 ℃, the constant-temperature shaking culture at the rotating speed of 40r/min is carried out to increase the ventilation volume, the pH value of the liquid culture medium is 6.0, the sugar concentration is 5%, and the yeast extract concentration is 2.0 g/L. Controlling the content of bacteriaAt 9.0 × 108CFU/mL is preferred (3.0 McLeod tube concentration).
Example 2 preparation method of compound Chinese medicine extract, wherein the ratio of each single Chinese medicine is as follows:
the traditional Chinese medicine compound extracting solution consists of the following components in parts by weight: 60-90 parts of antifebrile dichroa, 30-60 parts of tsaoko amomum fruits, 40-60 parts of humifuse euphorbia herb, 30-50 parts of purslane, 20-40 parts of copperleaf herb, 10-30 parts of Chinese silkvine, 10-30 parts of liquorice, 20-40 parts of dark plum, 20-50 parts of dried orange peel and 10-30 parts of medicated leaven.
The preparation method of the traditional Chinese medicine compound extract comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight of 6-10 times of the total weight of the traditional Chinese medicines, soaking for 30-60 minutes, decocting for 40-60 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, filtering to remove residues, and mixing the above 3 times of decoction filtrate.
Through experiments, the method comprises the following steps: after the traditional Chinese medicine compound extract prepared by the method is fermented, the anti-coccidiosis probiotic medicine can be obtained, and the treatment effect on the coccidiosis of the chicken is good, wherein the treatment effect of the prepared probiotic medicine is the best when the traditional Chinese medicine compound extract No. 1 is used as a raw material.
The following are some preferred formulas and preparation methods:
(1) the Chinese medicine compound extract No. 1 comprises the following components in parts by weight (unit: g): dichroa febrifuga 80, tsaoko amomum fruit 40, humifuse euphorbia 50, purslane 40, acalypha australis 30, Chinese chive 20, liquorice 20, dark plum 30, dried orange peel 40 and medicated leaven 20.
The preparation method of the Chinese herbal compound extract No. 1 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight of 8 times of the total weight of the traditional Chinese medicines, soaking for 60 minutes, decocting for 60 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
(2) The component proportion (unit: g) of the compound traditional Chinese medicine extract No. 2 is as follows: 60 parts of dichroa febrifuga, 30 parts of tsaoko amomum fruits, 50 parts of humifuse euphorbia herbs, 40 parts of purslane, 30 parts of copperleaf herbs, 10 parts of Chinese silkvine roots, 10 parts of liquorice, 20 parts of dark plums, 20 parts of dried tangerine peels and 10 parts of medicated leavens.
The preparation method of the Chinese herbal compound extract No. 2 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight 6 times of the total weight of the traditional Chinese medicines, soaking for 40 minutes, decocting for 40 minutes, and filtering to remove residues; adding 3 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 3 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
(3) The component proportion (unit: g) of the compound traditional Chinese medicine extract No. 3 is as follows: dichroa 90, tsaoko amomum fruit 60, humifuse euphorbia 60, purslane 50, acalypha australis 40, Chinese chive 30, liquorice 30, dark plum 40, dried orange peel 50 and medicated leaven 30.
The preparation method of the traditional Chinese medicine compound extract No. 3 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight being 10 times of the total weight of the traditional Chinese medicines, soaking for 60 minutes, decocting for 60 minutes, and filtering to remove residues; adding 5 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 5 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
(4) The Chinese herbal compound extract No. 4 comprises the following components in parts by weight (unit: g): dichroa febrifuga 80, tsaoko amomum fruit 40, humifuse euphorbia 40, purslane 30, acalypha australis 20, Chinese chive 20, liquorice 20, dark plum 30, dried orange peel 30 and medicated leaven 20.
The preparation method of the Chinese herbal compound extract No. 4 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight of 8 times of the total weight of the traditional Chinese medicines, soaking for 60 minutes, decocting for 60 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
(5) The Chinese herbal compound extract No. 5 comprises the following components in parts by weight (unit: g): 60 parts of dichroa febrifuga, 30 parts of tsaoko amomum fruits, 60 parts of humifuse euphorbia herbs, 50 parts of purslane, 40 parts of copperleaf herbs, 10 parts of Chinese silkvine roots, 20 parts of liquorice, 30 parts of dark plums, 40 parts of dried tangerine peels and 20 parts of medicated leavens.
The preparation method of the Chinese herbal compound extract No. 5 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight 9 times of the total weight of the traditional Chinese medicines, soaking for 50 minutes, decocting for 50 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
(6) The Chinese herbal compound extract No. 5 comprises the following components in parts by weight (unit: g): dichroa febrifuga 80, tsaoko amomum fruit 60, humifuse euphorbia 50, purslane 40, acalypha australis 30, Chinese chive 20, liquorice 10, dark plum 30, dried orange peel 40 and medicated leaven 20.
The preparation method of the compound traditional Chinese medicine extract No. 6 comprises the following steps: pulverizing each single traditional Chinese medicine by a superfine pulverizer, weighing the single traditional Chinese medicines according to the formula, adding deionized water with the weight being 10 times of the total weight of the traditional Chinese medicines, soaking for 60 minutes, decocting for 60 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 4 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues. Mixing the above 3 decoctions and filtrate.
Example 3 a method for preparing a fermentation filtrate, comprising the steps of:
taking the combined decoction filtrate prepared in each liter of example 2 as a unit, 200 g/L of maltose and 300g/L of wort and 200mL/L, MgSO of wort4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. Cooling to about 30 ℃, inoculating 80-100 mL/L of filtrate (the inoculum size is 8-10 percent, the percentage is volume percent) of the active saccharomyces cerevisiae culture prepared in the example 1, and culturing for 7-10 days by facultative anaerobic fermentation at 35 ℃ (the sealing film at the bottle mouth of the fermentation bottle is provided with air holes, is not completely sealed, is placed in an aseptic fermentation chamber, is cultured and observed by a shaking table at 40 r/min) until white mycoderm grows out on the liquid surface.
The following methods are preferred for preparing several better fermentation filtrates, and the combined decoction filtrate is preferably the compound traditional Chinese medicine extract No. 1:
(1) number 1 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 200 g/L of maltose and 100 mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. After cooling to about 30 ℃, 80 mL/L of the filtrate (inoculum size: 8%) of the active yeast culture prepared in example 1 was inoculated, and cultured at 35 ℃ for 7 days by facultative anaerobic fermentation (in a sterile fermentation room, cultured and observed by a shaker at 40 r/min) until a white mycoderm was grown on the liquid surface.
(2) Number 2 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 300g/L of maltose and 200mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. After cooling to about 30 ℃, 100 mL/L of the filtrate (10% inoculum) of the active yeast culture prepared in example 1 was inoculated, and cultured at 35 ℃ for 10 days by facultative anaerobic fermentation (in a sterile fermentation room, cultured and observed by a shaker at 40 r/min) until a white mycoderm was formed on the liquid surface.
(3) Number 3 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 200 g/L of maltose and 100 mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. After cooling to about 30 ℃, 100 mL/L of the filtrate (10% inoculum size) of the active yeast culture prepared in example 1 was inoculated, and cultured at 35 ℃ for 7 days by facultative anaerobic fermentation (in a sterile fermentation room, cultured and observed by a shaker at 40 r/min) until a white mycoderm was formed on the liquid surface.
(4) Number 4 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 300g/L of maltose and 200mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. Cooled to about 30 ℃, inoculated with 80 mL/L filtrate (inoculum size is 8%) of the active yeast culture prepared in example 1, cultured for 10 days in facultative anaerobic fermentation at 35 ℃ (observed in a sterile fermentation room, and cultured by a shaking table 40 r/min) until white mycoderm grows on the liquid surface.
(5) Number 5 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 300g/L of maltose and 200mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilizing at 121 deg.CFor 15 minutes. After cooling to about 30 ℃, 90 mL/L of the filtrate (inoculum size: 9%) of the active yeast culture prepared in example 1 was inoculated, and cultured at 35 ℃ for 7 days by facultative anaerobic fermentation (in a sterile fermentation room, cultured and observed by a shaker at 40 r/min) until a white mycoderm was grown on the liquid surface.
(6) Number 6 of fermentation filtrate: taking each liter of the combined decoction filtrate as a unit, 200 g/L of maltose and 200mL/L, MgSO of wort are added4 5.0 g/L、FeSO4 3.0 g/L. Sterilized at 121 ℃ for 15 minutes. Cooling to about 30 deg.C, inoculating 90 mL/L filtrate (inoculum size of 9%) of active yeast culture, and culturing at 35 deg.C for 7 days by facultative anaerobic fermentation (aseptic fermentation chamber, shaking table 40r/min culture observation) until white mycoderm grows on liquid surface.
Through experiments, the method comprises the following steps: the fermentation filtrate has good treatment effect on chicken coccidiosis, wherein the probiotic medicament prepared by taking the compound traditional Chinese medicine extract No. 1 as a fermentation raw material and adopting the formula and the preparation method of the fermentation filtrate No. 5 has the best treatment effect. Therefore, in the present invention, the following experiments will be explained with this formulation.
EXAMPLE 4 preparation and application of fermentation filtrates as drinking water agents and powders
(1) Preparing a drinking agent: and (3) removing the liquid surface mycoderm from the fermentation filtrate prepared in the example 3 to obtain the oral drinking water agent of the compound medicine. The invention preferably selects No. 5 fermentation filtrate to prepare the drinking water agent.
The using method comprises the following steps: the medicine is taken by drinking water on an empty stomach in the early morning, and the clinical treatment dosage is as follows: the drinking water of the chicks is added with 20-30 mL of drinking water agent medicine per liter, 1 time per day, and 5 days are 1 course of treatment.
(2) Preparation of the powder: the fermentation filtrate prepared in example 3 was concentrated by centrifugation (4 ℃, 5000-. Adding basic lyophilized protectant and protein stabilizer (w/w, mass percentage, 3-5% glycerol, 5-10% skimmed milk powder, 10-15% sucrose or mannitol, 10-15% beta-cyclodextrin or acacia gum, and the rest is drug precipitate, and adding together is 100%). Mixing, adjusting pH to 5.2-6.0, and balancing for 60 min. Pre-freezing at-20 deg.C for 12 h, and vacuum freeze-drying for 24 h to obtain compound medicinal powder.
The invention preferably prepares the fermentation filtrate No. 5 into powder and adds the powder to the medicine after mixing.
The using method comprises the following steps: the usage amount of the medicine powder is 200-600mg/kg of chicken weight, the medicine powder is mixed with feed and fed for 1 time/day, 5 days are 1 treatment course (about 250 g of chicks eat 50 g of feed approximately one day, the addition amount of 1 per thousand is calculated, 50 mg of powder is contained in 50 g of feed, and the conversion is 200 mg/kg of chicken weight, if the treatment dosage of acute morbidity is, the addition amount of the medicine powder can be increased to 3 g of powder added in each kilogram of chicken feed, and the conversion is 600mg/kg of chicken weight).
Example 5 acute toxicity test and median lethal dose determination
1 preliminary experiments
The preparation steps of the test drug are as follows:
(1) the active yeast culture was cultured according to the method of example 1 until the bacterial content reached 9.0X 108 CFU/mL；
(2) Preparing the compound traditional Chinese medicine extracting solution by the formula and the preparation method of the compound traditional Chinese medicine extracting solution No. 1 in the embodiment 2;
(3) the fermentation filtrate was prepared by the method of preparation of fermentation filtrate No. 5 in example 3;
(4) removing the liquid surface mycoderm from the fermentation filtrate to obtain the drinking water agent, and diluting by 3 times.
Test animals: 60 Kunming clean-grade mice, 18-22 g of weight of the mice, half of each mouse, 10 mice in each group and 6 groups. The test drugs for oral gavage were administered at different dosages (1.0, 2.0, 4.0, 8.0, 16.0, 20.0 mg/kg body weight) respectively, clinical symptoms were observed, the number of deaths was recorded, and the range of 0-100% of the death dose was determined.
2 acute toxicity test
Healthy mice were randomly divided into 5 dose groups (i.e. 4.0, 4.8, 5.76, 6.90, 8.30 mg/kg body weight dose) and 1 control group (blank control, no drug fill) with a 1:1.2 dose ratio between groups. 10 mice per group, half male and female, were gavaged orally and mice were observed for death after dosing and the results were recorded.
3 results and analysis
After administration, mice in medium and high dose groups have the phenomena of mental depression and activity reduction, and have no other obvious abnormal reactions. After 2 d, the diet and the water intake of the mice are increased to a certain degree, the activities are increased, the mental state is good, and abnormal reactions do not occur. 3-7 d after gastric lavage, the mice have good appetite and no toxic reaction.
4 conclusion
Calculating LD of medicine by improved Kouya method503679.8 mg/kg, with 95% confidence intervals: 2055.3-4315.6 mg/kg. According to the acute toxicity test regulation in the technical Specification for veterinary drug tests, mice are subjected to oral LD once in the classification standard of the acute toxicity of exogenous chemicals50Within the range of 500-5000 mg/kg, the drug is considered to be in the range of low toxicity-actual non-toxicity.
EXAMPLE 6 drug efficacy test against Eimeria tenella
1 materials and methods
1.1 test animals
120 Lingnan yellow chicks of 1 day old were purchased from Hualongan poultry breeding company, Lanzhou, Gansu province. The chick feed is purchased from Fuchang feed factory in Lanzhou city, and is not added with any coccidiostat. Feeding conditions and management: the standard experimental animal house (strictly disinfected and free from the environment polluted by chicken coccidian oocysts) of the Lanzhou livestock and veterinary research institute of Chinese academy of agricultural sciences is isolated for cage culture. Feeding the feed according to a conventional method, freely eating drinking water, feeding the feed to 14 days old, randomly grouping according to the weight, and feeding the feed in cages.
1.2 test drugs
The test drugs were prepared as follows:
(1) the active yeast culture was cultured according to the method of example 1 until the bacterial content reached 9.0X 108 CFU/mL；
(2) Preparing the compound traditional Chinese medicine extracting solution by the formula and the preparation method of the compound traditional Chinese medicine extracting solution No. 1 in the embodiment 2;
(3) the fermentation filtrate was prepared by the method of preparation of fermentation filtrate No. 5 in example 3;
(4) the fermentation filtrate prepared in example 3 was concentrated by centrifugation (4 ℃ C., 5000 rpm, 10 min), and the drug complex precipitate at the bottom of the tube was collected. Adding basic lyophilized protectant and protein stabilizer (w/w, mass percent, 7% skimmed milk powder, 4% glycerol, 12% sucrose, 10% beta-cyclodextrin, 67% drug precipitate) into the precipitate, mixing, adjusting pH to 6.0, and balancing for 60 min. Pre-freezing at-20 deg.C for 12 h, and vacuum freeze-drying for 24 h to obtain compound medicinal powder.
The using method comprises the following steps: mixing with feed, and feeding for 1 time/day, 5 days as 1 course of treatment. The test is divided into 6 drug dose groups, the addition amount of the powder is respectively 50, 100, 200, 300, 400 and 500 mg/kg of the weight of the chicken, and the powder is mixed with feed for administration and is continuously administered for 7 days.
Control drugs: diclazuril, purchased from the pharmaceutical industry ltd of agricultural high-tech animals in south Jiangsu, was administered at a dose of 1.0 mg/kg, as recommended by the manufacturer's instructions.
1.3 test insect strains
Eimeria tenella (mild resistance to nicarbazin, moderate resistance to halofuginone and croissant, severe resistance to furazolidone, clopyralid, maduramicin, lasalomycin, salinomycin, monensin) was provided by the lanzhou veterinary institute, the chinese academy of agricultural sciences.
1.4 anticoccidial test
1.4.1 test animals grouping and dosing
When the chicks were raised to 12 days of age, the test chickens were fecal checked for two consecutive days and after confirming the absence of coccidial infection, the test was started at 14 days of age. Selecting 90 healthy coccidian-free chicks, weighing the chicks one by one, selecting the chicks with similar weight, and randomly dividing the chicks into 9 groups of 10 chicks each. 6 drug dose groups (feed mixed with drug), diclazuril control group (1.0 mg/kg mixed with drug), infection control group (red control group) and healthy control group (white control group). Except for the white control group, each chicken of the other infection groups was orally inoculated with 6 ten thousand of completely sporulated oocysts (oocysts were eimeria tenella sporulated oocysts), and administration was started 24 hours after the inoculation and continued for 7 days.
1.4.2 clinical and pathological dissection observations
After the inoculation of Eimeria tenella, the disease and clinical manifestations of the groups of test chickens were observed and recorded. On day 3 post-infection, feces were examined daily and bloody stools were recorded until the end of the test. On days 5-8 post-infection, feces were collected and weighed daily and Oocysts per gram of feces (OPG) were counted using a Myworm egg counting method (saturated saline float method). After the test on the 8 th day, each group of live chickens was weighed individually, and the weight gain of each group was calculated. Then killing all chicken ceca, collecting all groups of chicken ceca, longitudinally cutting off the chicken ceca, and scoring the lesions of each group of chicken ceca according to a Johnson & Reid (1970) lesion scoring method. Meanwhile, the cecal contents and mucosal epithelial tissues of each group were collected, and the amount of cecal oocysts was counted.
1.5 evaluation indexes and standards for therapeutic effects of drugs
1.5.1 weight gain = (average body weight after end of test-average body weight at start of test)/average body weight at start of test × 100%.
1.5.2 relative rate of weight gain = (mean rate of weight gain in infected test group ÷ mean rate of weight gain in healthy control group) × 100%.
1.5.3 lesion scoring the chickens in each group were observed daily for mental, appetite, faeces, mortality etc. and the results were recorded. The chickens that died during the experiment were autopsied and observed for pathological changes with reference to the Johnson & Reid evaluation criteria.
The lesion score is in the range of 0-4 +, 0 represents no abnormality, 4+ represents the most serious lesion, cecal bleeding is serious, the degree of swelling is high, and the intestinal cavity is filled with blood coagulation and mucous membrane fragments. In the test, the intestinal lesion scores of the chickens which died due to coccidiosis were all scored as 4.
Lesion value = mean lesion score per group of chickens x 10.
1.5.4 score of feces is based on Morehouse and Baron (1970) evaluation criteria, and the score of abnormal feces is in the range of 0-4 +, 0 indicates no abnormality, 4+ indicates the most serious abnormal thin feces, and feces have blood and mucus.
1.5.5 survival rate: survival (%) = (number of surviving chickens/number of starting chickens) × 100%.
1.5.6 oocyst value: the amount of oocysts in the infected control group was 100% (40 min) as calculated by the okadah value method.
1.5.7 Anticoccidial index (ACI)
The anticoccidial index ACI was calculated from the relative weight gain, survival rate, lesion value and oocyst value of each group of test chickens according to the method of Merck, USA.
Determining the curative effect of the medicine by taking ACI as a standard, wherein the ACI is not more than 120 and is ineffective; ACI is low efficiency between 120 and 160; ACI is between 160 and 180 for medium effect; ACI > 180 is highly effective.
ACI = (relative rate of weight gain + survival) × 100- (lesion value + oocyst value).
2 results of the test
2.1 clinical symptoms and bloody stool
In 3 d after coccidium inoculation, except for the white control group, the other infection test group chickens all show different degrees of clinical symptoms, such as reduced feed intake, plump feathers, drooping wings, necking down, closed eyes and sleeping, unwilling to walk, dislocating and standing, discharging blood and defecation and the like.
In 4 d and 5 d after coccidiosis inoculation, except for the white control group feces score of 0, the red control group chickens still show the symptom of discharging a large amount of bloody stool (feces score of 4), the bloody stool of all the drug dose groups is obviously reduced, and the mental condition and the food consumption are obviously improved (feces score of 3).
2.2 pathological anatomical changes
Pathological anatomy shows that the visceral organs of all white control test chickens are normal. The red control group tested chickens, with significant cecal and duodenal swelling, contained large amounts of blood sample contents. In all drug dose groups, swelling and blood sample contents also appear in the caecum and the duodenum of the tested chicken, but the pathological change degree is obviously reduced compared with that of an infected control group, and other visceral organs have no obvious ocular pathological change. The values of the lesions are shown in Table 1.
2.3 anticoccidial Effect
The ACI was calculated from the indexes such as the relative weight gain rate, the survival rate, the oocyst value, and the lesion value, and as can be seen from Table 1, the ACI values of the drug dose groups of the experimental drugs were 89.7, 128.2, 155.8, 163.7, 165.7, and 157.0, respectively, which were higher than those of the red control group (ACI of 68.7); 200. the ACI values of the 300, 400 and 500 mg/kg chicken weight dose groups were all higher than that of the diclazuril drug group (ACI value of 134.8).
TABLE 1 therapeutic results of the fermentation filtrate No. 5 powder against Eimeria tenella
Figure DEST_PATH_IMAGE002
3. In conclusion:
the medicine is administered by mixing with 200, 300, 400 and 500 mg/kg chicken weight dose of feed, and can improve weight gain rate of test chicken and artificial infectionE.tenellaThe disease test chicken only has good treatment effect, wherein the anticoccidial index of 300 and 400 mg/kg chicken weight dose groups is higher than 160, belongs to medium curative effect and is obviously higher than that of a diclazuril drug control group and a red control group.
In view of the fact that the test has the conditions of strong toxicity of insect strains and large insect attack amount, the anticoccidial curative effect observation test proves that the test medicament can effectively prevent the chicken coccidiosis under the artificial infection condition and has better anticoccidial curative effect than the chemical medicament diclazuril. The traditional Chinese medicine extract disclosed by the invention is low in toxicity, low in residue, large in safety range and ideal in curative effect comparison test effect after being subjected to anaerobic fermentation by using an active yeast culture, and is expected to become a novel coccidian-resistant medicine.
Example 7 comparison of Pre-and post-fermentation Effect experiments on probiotic drugs according to the invention
Acute toxicity results demonstrate that the fermented drug of the present invention can alleviate the symptoms of diarrhea in mice, drug LD, compared to the unfermented drug50The toxicity level is obviously reduced when the concentration is increased from 2032.2 mg/kg to 3679.8 mg/kg. Secondly, the fermented medicine of the invention has no abnormal pathological change in histopathological and histological examination of the tissue organs, and no damage, blood stasis and enlargement of the liver and the kidney of the mouse. The liver lobule profile under the light microscope is clear, and the liver cells have no deformation and necrosis. The mice which are clinically visible after the non-fermented medicine is infused with the medicineMental depression and pile-tying reaction, the liver is hyperemia and congestion phenomena can be seen through autopsy, liver cells are changed in a diffuse manner under a light mirror, hepatic sinuses are obviously congested and dilated, the liver cells are deformed and necrotized, and kidneys are shown as cell swelling, granule degeneration and transparent degeneration.
The total polysaccharide content of the fermented medicine extract is increased by 3.57 times compared with that of the medicine extract which is not fermented (measured by a phenol-sulfuric acid method), and the high performance liquid chromatography and the medicine stability detection result show that the contents of dichroin and dichroin A are respectively improved by 19 percent and 16 percent. The chronic toxicity test proves that compared with the unfermented drug, the fermented drug can obviously improve the average daily feed intake (from 11.3 g/d to 15.2 g/d) and the feed utilization rate (the material weight ratio is reduced from 9.9 to 5.8) of the rat, improve the average daily gain (from 1.33 g/d to 2.67 g/d), and has better effects of promoting growth and weight. The symptoms of diarrhea and hematochezia of rats are not seen in the eye, and the safety of medication is further improved. In clinical treatment experiments, compared with the non-fermented medicine, the caecum and the duodenum of the test chicken taking the fermented medicine have no obvious swelling and blood sample contents, the intestinal mucosa has no obvious hyperemia and hemorrhage phenomenon, and part of the chicken have the splenic lymph nodes (which proves that the intestinal mucosa immunity is improved). The relative weight gain rate of the chicken is increased by 11 percentage points, and the coccidian resistance index is increased from 142.3 to 165.7. )
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. 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.

Claims (8)

1. A preparation method of a chicken coccidiosis resistant probiotic medicine is characterized by comprising the following steps: the probiotic medicament is prepared from a traditional Chinese medicine compound extract through saccharomyces cerevisiae facultative anaerobic fermentation; the traditional Chinese medicine compound extracting solution is prepared by mixing the following raw materials in parts by mass and then carrying out water extraction: 60-90 parts of antifebrile dichroa, 30-60 parts of tsaoko amomum fruits, 40-60 parts of humifuse euphorbia herb, 30-50 parts of purslane, 20-40 parts of copperleaf herb, 10-30 parts of Chinese silkvine, 10-30 parts of liquorice, 20-40 parts of dark plum, 20-50 parts of dried orange peel and 10-30 parts of medicated leaven.
2. The method for preparing a probiotic medicament against chicken coccidiosis according to claim 1, wherein the method comprises the following steps: the traditional Chinese medicine compound extracting solution is prepared by mixing the following raw materials in parts by mass and then carrying out water extraction: 80 parts of antifebrile dichroa, 40 parts of tsaoko amomum fruits, 50 parts of humifuse euphorbia herbs, 40 parts of purslane, 30 parts of copperleaf herbs, 20 parts of Chinese silkvine, 20 parts of liquorice, 30 parts of dark plums, 40 parts of dried orange peels and 20 parts of medicated leavens.
3. The method for preparing a probiotic medicament against chicken coccidia according to claim 1 or 2, characterized in that: the preparation method of the traditional Chinese medicine compound extracting solution comprises the following steps: crushing the raw materials, weighing the raw materials according to the formula, adding deionized water with the weight 6-10 times of the total weight of the traditional Chinese medicines, soaking for 30-60 minutes, decocting for 40-60 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, and filtering to remove residues; adding 3-5 times of deionized water into the filter residue, decocting for 40 minutes, filtering to remove residues, and mixing the above 3 times of decoction filtrate.
4. The method for preparing a probiotic medicament against chicken coccidia according to claim 1 or 2, characterized in that: the fermentation method of the traditional Chinese medicine compound extracting solution comprises the following steps: 200 g/L maltose and 200mL/L, MgSO malt extract are added into the traditional Chinese medicine compound extracting solution4 5.0g/L、FeSO43.0g/L, sterilizing, cooling to 28-32 deg.C, inoculating 8-10% of Saccharomyces cerevisiae, and performing facultative anaerobic fermentation culture in 35 deg.C sterile fermentation chamber by shaking table until white mycoderm grows on liquid surface.
5. A probiotic medicament against coccidiosis prepared by the method of any one of claims 1 to 4.
6. The anti-coccidiosis probiotic drug of claim 5, wherein: removing the liquid surface mycoderm from the fermented filtrate obtained by fermentation to obtain the oral drinking water agent of the probiotic medicine.
7. Use of the anti-coccidial probiotic of any one of claims 5 to 6 in the preparation of a medicament or feed additive for the treatment of coccidiosis in chickens.
8. Use according to claim 7, characterized in that: the chicken coccidiosis is chicken coccidiosis caused by Eimeria tenella.
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