CN106822890A - A kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets and its preparation method and application - Google Patents

A kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets and its preparation method and application Download PDF

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Publication number
CN106822890A
CN106822890A CN201710037459.2A CN201710037459A CN106822890A CN 106822890 A CN106822890 A CN 106822890A CN 201710037459 A CN201710037459 A CN 201710037459A CN 106822890 A CN106822890 A CN 106822890A
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dry powder
yolk
igy
parts
pharmaceutical composition
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CN106822890B (en
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王玲
刘宇
杨峰
魏小娟
郭文柱
罗永江
张彬
杨珍
周绪正
罗金印
牛建荣
郭爱民
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Lanzhou Institute of Animal Husbandry and Veterinary Medicine CAAS
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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
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/57—Birds; Materials from birds, e.g. eggs, feathers, egg white, egg yolk or endothelium corneum gigeriae galli
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/63—Arthropods
    • 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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A61K36/064—Saccharomycetales, e.g. baker's yeast
    • 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)
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • 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)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/288—Taraxacum (dandelion)
    • 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
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/489—Sophora, e.g. necklacepod or mamani
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/68—Plantaginaceae (Plantain Family)
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    • 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)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A—HUMAN NECESSITIES
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    • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
    • A61K36/718—Coptis (goldthread)
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
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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)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/739—Sanguisorba (burnet)
    • 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)
    • A61K36/756—Phellodendron, e.g. corktree
    • 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)
    • A61K36/78—Saururaceae (Lizard's-tail family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

Abstract

The present invention provides a kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets, and described pharmaceutical composition is made up of following each component：It is anti-E.ColiIgY Yolk antibodies dry powder, anti-TGE PED PoRV IgY Yolk antibodies dry powder, traditional Chinese medicine compound extract dry powder, powdery yolk and probiotics yeast bacterium dry powder.The present invention also provides its preparation method and the application in the medicine for preparing preventing and treating diarrhea of weaned piglets.Pharmaceutical composition of the invention can have certain development and application values as the medicine and feed addictive of the preventing and treating diarrhea of weaned piglets of high-quality.Preparation method of the invention, selection is convenient, extraction efficiency is high, advantage of lower cost, it is easy to form large-scale production.

Description

A kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets and preparation method thereof and Using
Technical field
The invention belongs to veterinary drug technical field, it is related to a kind of medicine for preventing and treating grice diarrhoea, and in particular to one kind is for preventing Control pharmaceutical composition of diarrhea of weaned piglets and its preparation method and application.
Background technology
Grice diarrhoea is a kind of typical multifactor property disease, causes the cause of disease species of grice diarrhoea various, and characteristic of disease is complicated, Often by various cause of disease mixed infections.Grice diarrhoea is the important original for causing piglet growth retardation and piglet high mortality for a long time Cause, the preventing and treating of grice diarrhoea is always a noticeable worldwide problem.The cause of disease of grice diarrhoea is caused to be summarized as non-biography Metachromia is suffered from diarrhoea and the major class of infectious diarrhea two.The non-infectious diarrhoea of piglet often because piglet digestive function is not complete, daily ration antigen allergy, Trophic factors lacks, feeding and management and stress factor cause the reduction of piglet resistance and diarrhoea occur.Infectious diarrhea includes thin Bacterium property and virus diarrhea, are taken turns by enteropathogenic E. Coli, transmissible gastro-enteritis virus, Porcine epidemic diarrhea virus and pig more Shape virus causes.And grice diarrhoea is prevented and treated at present, and comprehensive preventive health measures is mainly taken, on the basis of good feeding and management, pin To the correct from being the key problem in technology of preventing and treating grice diarrhoea of property vaccine and sensitive medicaments.The integrated control skill of bacterial diarrhea Art, mainly takes comprehensive preventive health measures in terms of administration of health, vaccine prevention and medical treatment three is raised.Because virus diarrhea does not have There is specific medicament, mainly prevented and treated by strengthening feeding and management and vaccine prevention at present, additionally, heteropathy, oral micro- life State preparation, from sensitive antibiotic etc. prevent bacterium scabies secondary infection can as reduce virus diarrhea supplementary means.Therefore, Select, specificity height sensitive to pathogen, fungistatic effect is strong, good stability newtype drug is particularly important, and is foster from now on The important R＆D direction that pig industry develops in a healthy way.
After ablactation being typically occurred in due to grice diarrhoea（28 ages in days）Piglet, therefore, culturist is more likely to susceptible The with strong points, bioavilability of the interior use of age in days is high, high safety medicine, is prevented and is controlled by easily oral route Treat, with this make up vaccine injection presence using inconvenience, side effect big, complex process, half-life short, preserve the deficiencies such as inconvenience Place.
The content of the invention
In order to solve problems of the prior art, the invention provides one kind for preventing and treating diarrhea of weaned piglets medicine The preparation method and application of composition.The pharmaceutical composition is by special yolk antibody IgY, probiotics, Traditional Chinese medicine compound composition And powdery yolk composition.
The mechanism of action of pharmaceutical composition of the invention is：
Targeting drug delivery system can make drug accumulation be positioned at pathological tissues, organ, cell or intracellular structure, the target for thus carrying out Can significantly improve the drug concentration of therapentic part to treatment, so as to reduce dosage, reduce toxic and side effect of the medicine to whole body, And reduce medical expense.Special yolk antibody IgY is mainly using the principle of antigen-antibody reaction to build targeted drug, Specificity is followed the trail of and caused in enteron aisle diarrhoea common causative, is oriented by passive immunity approach and is killed pathogenic microorganisms, reduces infection Degree, with targeting it is strong, have no toxic side effect, do not produce drug resistance the features such as.Simultaneously utilize with bacteriostasis antibiosis, eliminating dampness and heat, Dampness removing antidiarrheal for principle prescription Chinese medicine composition, to the further anti-inflammation of pathogen, and have concurrently raising pigling immunity and Effect of premunition, and excitant is small, low toxicity noresidue, be not likely to produce drug resistance, security is higher.Probiotics yeast bacterium (Saccharomyces cerevisiae) containing profuse protein, its culture is rich in vitamin, mineral matter, digestion Enzyme, somatomedin, and peptide, various essential amino acids and essential fatty acid, are after another after animal protein and vegetable protein One class key protein is originated.These special nutrient compositions can stimulating immune system defence capability, reduce enteral noxious material Generation, there is facilitation to body's immunity, thus saccharomycete and its fermented product can as the feed addictive of high-quality, There are certain development and application values.By giving full play to the synergy between said medicine, to reach for various etiologies Preventive and therapeutic effect, reduce grice diarrhoea morbidity and mortality, improve culture benefit.Pharmaceutical composition effect of the invention is better than Antibiotic therapy alone or simple probiotic products, and simple yolk antibody IgY product, are preventing and are treating grice diarrhoea Aspect has preferable targeted therapy and a duration of efficacy action character long, and do not produce drug resistance, have no toxic side effect, nothing Residual, it is easy to product scale and production cost is relatively low.
First purpose of the invention is to provide a kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets, the medicine Composition is made up of following each component：It is anti-E.Coli- IgY Yolk antibodies dry powder, anti-TGE-PED-PoRV-IgY Yolk antibodies are done Powder, traditional Chinese medicine compound extract dry powder, powdery yolk and probiotics yeast bacterium dry powder；
The traditional Chinese medicine compound extract dry powder is prepared from by each component of following mass fraction：Kuh-seng 10-30 parts, garden burnet 10- 30 parts, coptis 10-20 parts, golden cypress 10-20 parts, Chinese gall 10-20 parts, dandelion 10-20 parts, wolf's milk 10-20 parts, cordate houttuynia 5-15 parts, hairyvein agrimony 5-15 parts, purslane 5-15 parts, Asiatic plantain 5-15 parts, polygonum perfoliatum 2-15 parts, milfoil 2-15 parts, Radix Glycyrrhizae 5- 15 parts.
Preferably, the traditional Chinese medicine compound extract dry powder is prepared from by each component of following mass fraction：Kuh-seng 15 Part, 15 parts of garden burnet, 12 parts of the coptis, 12 parts of golden cypress, 12 parts of Chinese gall, 15 parts of dandelion, 10 parts of wolf's milk, 10 parts of cordate houttuynia, celestial being 12 parts of crane grass, 12 parts of purslane, 10 parts of Asiatic plantain, 8 parts of polygonum perfoliatum, 6 parts of milfoil, 6 parts of Radix Glycyrrhizae.
Preferably, in described pharmaceutical composition, the probiotics yeast bacterium dry powder is 0.5 × 108--5.0×108 CFU/g；It is described anti-E.Coli- IgY Yolk antibodies dry powder, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder, Chinese medicine compound prescription are carried It is 100% to take the mass percent sum of liquid dry powder and powdery yolk, wherein, resistE.Coli- IgY Yolk antibody dry powder 2.5-10 %, Anti- TGE-PED-PoRV-IgY Yolk antibodies dry powder 2.5-10 %, traditional Chinese medicine compound extract dry powder 60-90 %, powdery yolk 5-30%；
Preferably, described anti-E.Coli- IgY Yolk antibodies dry powder is 5%, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder It is 5%, traditional Chinese medicine compound extract dry powder is 70%, and powdery yolk is 20%.
Preferably, the probiotics yeast bacterium dry powder is prepared from by saccharomyces cerevisiae, bacterial content is 1.0-2.0 × 109 CFU/g。
Preferably, described anti-E.ColiThe preparation method of-IgY Yolk antibody dry powder is：Using coli somatic egg White antigen immune thing is immunized to healthy laying hen, collects egg, and separating special yolk using water dilution method-freezing resists Body IgY, supernatant is anti-E.Coli- IgY Yolk antibodies, by supernatant freeze-drying after, resistedE.Coli- IgY yolk Antibody dry powder.
Preferably, the preparation method of the anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder is：By pig transmissible stomach Enteritis-pig epidemic diarrhea-porcine rotavirus G5 type trigeminal live vaccine immune health laying hens, collects egg, adopts dilute with water Method-freezing separates special yolk antibody IgY, and supernatant is anti-TGE-PED-PoRV-IgY Yolk antibodies, by supernatant After freeze-drying, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder is obtained.
Preferably, the powdery yolk is by resistingE.ColiThe solid-state powdery yolk and anti-TGE-PED-PoRV- in-IgY antibody source The weight portions such as the solid-state powdery yolk in IgY antibody source are mixed；
It is described anti-E.ColiThe preparation method of the solid-state powdery yolk in-IgY antibody source is：Exempted from using coli somatic proteantigen Epidemic disease thing is immunized to healthy laying hen）, egg is collected, separated using water dilution method-freezing, the spraying of yolk sediment is dry It is dry, resistedE.ColiThe solid-state powdery yolk in-IgY antibody source；
The preparation method of the solid-state powdery yolk in the anti-TGE-PED-PoRV-IgY antibody source is：By transmissible gastroenteritis of swine-pig Epidemic diarrhea-porcine rotavirus G5 type trigeminal live vaccine immune health laying hens, collects egg, using water dilution method-freezing Method is separated, and by the spray drying of yolk sediment, obtains the solid-state powdery yolk in anti-TGE-PED-PoRV-IgY antibody source.
Preferably, the preparation method of the traditional Chinese medicine compound extract dry powder is：After each component of formula ratio is crushed, plus 3-10 times of decocting is boiled 3 times, each 1-4 hours, merging filtrate, vacuum-concentrcted, concentrate spray drying, obtains Chinese medicine compound prescription Extract solution dry powder.
Second object of the present invention is to provide the preparation method of aforementioned pharmaceutical compositions, first will be anti-E.Coli- IgY yolk Antibody dry powder and anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder carry out waiting weight portion to mix, then by mixed Yolk antibody Dry powder mixes with traditional Chinese medicine compound extract dry powder and powdery yolk according to formula ratio, and adds probiotics yeast bacterium dry powder to be mixed, It is made pharmaceutical composition.
Third object of the present invention be to provide aforementioned pharmaceutical compositions prepare preventing and treating diarrhea of weaned piglets medicine or Application in feed addictive.
When preparing medicine, operated according to formulation method of the prior art, auxiliary material commonly used in the art can be added. Such as medicine can be directly added into suitable quantity of water, allow piglet to drink.
Beneficial effects of the present invention are：
（1）Pharmaceutical composition prepared by the present invention is matched somebody with somebody by by yolk antibody IgY with probiotics and Traditional Chinese medicine compound composition Than, coupling, can by special yolk antibody IgY orient catch grice diarrhoea etiology, prepare directed toward bacteria, virus beast Use target therapeutic agent.Secondly, Chinese medicine compound prescription active principle in addition to there is anti-inflammation, eliminating dampness and heat, dampness removing antidiarrheal, The characteristics of also having raising diarrhoea pigling immunity and resistance against diseases concurrently, and excitant is small, low toxicity noresidue, be not likely to produce resistance Property, it is safe.Probiotics yeast bacterium can stimulate the defence capability of piglet immunological system, reduce the generation of noxious material in enteron aisle, There is facilitation to body's immunity.Therefore, pharmaceutical composition of the invention has to the grice diarrhoea that bacterium, virus cause There is good targeted therapy and prevention effect, the double effectses with prevention and health care and disease treatment.
（2）The special yolk antibody IgY of preparation, acts on good targeted therapy, can be by passive immunity approach Specifically, orientation is killed causes the Escherichia coli of grice diarrhoea, can be used for the piglet abdomen that clinical prevention is caused by Escherichia coli Rush down, more afterwards diarrhoea piglet no longer recur, and have stable in properties, high specificity, noresidue concurrently, without advantages such as the resistances to the action of a drug.Meanwhile, energy Enough specific neutralization transmissible gastroenteritis of swine, pig epidemic diarrhea and porcine rotavirus, can be clinically used for treatment and are passed by pig Viral grice diarrhoea caused by the independent infection of metachromia gastroenteritis, pig epidemic diarrhea and porcine rotavirus or mixed infection.Make The delactational piglets of the above-mentioned etiology of easy infection can not only be cured with the pharmaceutical composition prepared in the present invention, while to same group not The delactational piglets of above-mentioned cause of disease are infected using its can be made to obtain passive immunity.
（3）Pharmaceutical composition prepared by the present invention, can avoid vaccinating in the past wastes time and energy（Such as epidemic disease it is occurred frequently, frequency Need to be used for multiple times during hair and just can reach effect）, emergency reaction is strong, vaccine preparation process complexity causes producing cost to increase Plus, in egg all multi-nutrients it is destroyed and the problems such as cause the wasting of resources.The pharmaceutical composition prepared using the present invention is had Help recover profitable strain in enteron aisle, Swine herd production performance can be significantly improved, with preferable growth promoting function.Present invention system Standby pharmaceutical composition has the advantages that easy to use, with low cost, long shelf-life, using rear instant effect, has no toxic side effect, no The appearance of drug-fast bacteria strain can be caused, significantly reduce because inject using caused by epidemic disease propagate and it is various seriously stress be anti- Should, suitable large-scale promotion.
（4）Pharmaceutical composition disclosed by the invention, is had by special yolk antibody IgY, probiotics, Traditional Chinese medicine compound composition Effect component and yolk antibody IgY byproduct --- powdery yolk is constituted, and nutritional ingredient is high, and palatability preferably, can be prepared into any one The oral formulations type for being easy to diarrhoea piglet application is planted, powder form can be prepared into and mixed for veterinary clinic drinking water administration or feed Clothes administration, or feed addictive is prepared into for preventing and treating diarrhea of weaned piglets.
Chinese medicine composition of the invention according to Chinese veterinarian's medicine and modern traditional Chinese medicine theory, with antibacterial bacteriostatic, heat-clearing and damp-drying drug, Gut purge antidiarrheal is principle, is formed according to certain weight portion prescription, and the Chinese medicine composition includes kuh-seng, garden burnet, the coptis, Huang Cypress, Chinese gall, dandelion, wolf's milk, cordate houttuynia, hairyvein agrimony, purslane, Asiatic plantain, polygonum perfoliatum, milfoil and Radix Glycyrrhizae, each single The pharmacological action of Chinese medicine is specific as follows：
Kuh-seng：Classification systemSophora flavescens var. flavescens.Nature and flavor：Bitter, it is cold in nature.Enter the heart, liver, Kidney, intestine and small intestine, bladder warp.Indication：Heat-clearing and damp-drying drug, wind dispelling insecticide.Main damp-heat dysentery, hematochezia, jaundice, urine is not Profit, oedema, under band, mange, leprosy, pruitus, wet poison sore.Kuh-seng decoction is to staphylococcus aureus, shigella dysenteriae etc. There is stronger bacteriostasis, have suppression to trichophyton, Trichophyton concentricum, Trichophyton schoenleini, Du small gemma tinea bacterium of big belly difficult to understand etc. Make and use.
Garden burnet：Classification systemSanguisorba officinalis L., medicine attribute：It is bitter, sour, puckery, it is slightly cold.Return liver, Large intestine channel.Indication：Cooling blood and hemostasis, purging intense heat and detonicating.Experiment in vitro is proved：The decocting liquid of garden burnet 100% is to typhoid bacillus, brain Film inflammation diplococcus, shigella flexneri, bacterium sonnei, beta hemolytic streptococcus, staphylococcus aureus, the double balls of pneumonia Bacterium, corynebacterium diphtheriae, Escherichia coli, hay bacillus, typhoid bacillus, paratyphosum Bacterium, Pseudomonas aeruginosa, comma bacillus and M. tuberculosis Bacillus has different degrees of inhibitory action.Also there are different degrees of inhibitory action decoctions to some fungies in 0.50 mg/mL When it is effective to Asia influenza A virus.
The coptis：Classification systemCoptis chinensis Franch, nature and flavor：It is bitter, cold, nontoxic.The thoughts of returning home, spleen, stomach, liver, Courage, large intestine channel.Indication：Heat-clearing and damp-drying drug, purging intense heat and detonicating.In vitro to staphylococcus aureus, hemolytic streptococcus, pneumonia ball Bacterium, diplococcus meningitidis, shigella dysenteriae, bacillus anthracis etc. have preferable antibacterial action, also with preferable antivirus action, Such as suppress influenza virus, hepatitis B, the protozoons such as Amoeba, Trichomonas vaginalis, trypanosome are may also suppress in vitro.
Golden cypress：Classification systemCortex Phellodendri Chinensis, nature and flavor：It is bitter, cold.Return kidney channel, bladder warp, Large intestine channel.Indication：It is clearing heat and detoxicating, heat-clearing and damp-drying drug.Cure mainly damp-heat dysentery, diarrhea, jaundice etc..In vitro test is to golden yellow Portugal Grape coccus, pneumococcus, corynebacterium diphtheriae, Streptococcus viridans, shigella dysenteriae（Except Song Nei Shi）, hemolytic streptococcus, meninx Scorching coccus, comma bacillus, bacillus anthracis are effective or have stronger inhibitory action；To hay bacillus, Bordetella pertussis, lockjaw Bacillus also has inhibitory action.
Chinese gall：Classification systemRhus chinensis Mill., meridian distribution of property and flavor：It is sour, puckery, it is cold in nature, return lung, large intestine, Kidney channel.Effect：Astringe the lung, hidroschesis, puckery intestines, controlling nocturnal emission with astringent drugs, hemostasis, removing toxic substances.Cure mainly：Chronic cough of deficiency lung, spontaneous sweating, protracted dysentery chronic diarrhea is various Bleeding etc..Chinese gall decoction has suppression to make to staphylococcus aureus, bacterium flexneri, hog cholera bacillus and Escherichia coli With.This product can penetrate intestinal tissue, suppress intestinal tube smooth muscle and wriggle and have anti-diarrhea effect.
Dandelion：Classification systemHerba Taraxaci.Nature and flavor：It is sweet, slight bitter, cold.Indication：It is clearing heat and detoxicating, detumescence Dissipating bind.Cure mainly：Acute mastitis is swollen and ache, sore Huang serious case of furuncle has good effect.There is stronger killing to staphylococcus aureus and hemolytic streptococcus Bacterium acts on, and has certain bactericidal action to Pseudomonas aeruginosa, proteus, shigella dysenteriae etc..
Wolf's milk：Classification systemEuphorbia humifusa Willd、Euphorbia maculata L..Nature and flavor： It is pungent, flat.Return lung, liver, stomach, large intestine, bladder warp.Effect：Clearing heat and promoting diuresis, subdhing swelling and detoxicating, cooling blood and hemostasis.Experiment in vitro shows：Entirely Grass has very strong antibacterial action, and antimicrobial spectrum is also very wide, and the fresh juice of wolf's milk, decoction and water boiling concentration ethanol extract for Various pathogenic coccuses and bacillus have obvious bacteriostasis.
Cordate houttuynia：Classification systemHouttuynia cordata Thunb., taste is pungent is cold, return lung, bladder, large intestine Through.Energy is clearing heat and detoxicating, sore, promoting urination and removing dampness, clearing away heat to cure dysentery, stomach strengthening and digestion promoting are treated in detumescence, is trouble with real heat, heat toxin, damp evil, disease heat is controlled Lung carbuncle, sore swollen toxin, hemorrhoidal hemorrhage, retention of heat in the spleen and stomach etc..Modern pharmacology experiment shows that this product has antibacterial, antiviral, raising Immunity of organisms, diuresis etc. are acted on.Secondly, to various microorganisms（Especially saccharomycete and mould）There is inhibitory action, to molten The streptococcus of courage and uprightness, staphylococcus aureus, Bacillus influenzae, micrococcus catarrhalis, pneumococcus have obvious inhibitory action, to large intestine Bacillus, shigella dysenteriae, typhoid bacillus also have effect.
Hairyvein agrimony：Classification systemAgrimonia pilosa Ldb., slightly warm in nature, taste is pungent is puckery, temperature, nontoxic.Return lung, liver, spleen Through.The puckery property held back of the product, can relieving diarrhea with astringents stop dysentery, because of the product mild in medicine property and, and can qi-restoratives, and can stop blooding, thus for bloody flux and Prolonged illness rush down dysentery is particularly suitable.The hot water or ethanol extract of hairyvein agrimony are in vitro to hay bacillus, staphylococcus aureus, large intestine Bacillus, Pseudomonas aeruginosa, shigella flexneri and typhoid bacillus etc. have inhibitory action.
Purslane：Classification systemPortulaca oleracea L., meridian distribution of property and flavor：It is cold in nature, sweet acid, enter the heart, liver, spleen, Large intestine channel.Effect：Clearing heat and detoxicating, Li Shui dries, and dissipates blood detumescence, dust-collection sterilization, anti-inflammatory analgetic, blood-stanching and blood-cooling.Cure mainly dysentery Disease, enteritis, ephritis, postpartum uterine hemorrhage is had blood in stool, the illness such as mastitis.Its ethanol extract to Escherichia coli, proteus, Shigella dysenteriae, typhoid fever, paratyphosum Bacterium have the inhibitory action of height.To staphylococcus aureus, fungi such as big belly microspore Du difficult to understand Tinea bacterium, tubercle bacillus also have different degrees of inhibitory action.There is slight inhibitory action to Pseudomonas aeruginosa.Experiment shows, in test tube It is interior（1：4）There is bactericidal action to shigella dysenteriae.Decoction in 18.75-37.5 mg/mL concentration, will is congratulated, in Song, Si Shi and Expense Shigella flexneri has inhibitory action.
Asiatic plantain：Classification systemPlantago asiatica, nature and flavor：It is sweet, light, cold nature.Return through：Return lung warp, Liver Channel, Kidney channel, bladder warp.Function：Reducing fever and causing diuresis, excreting dampness antidiarrheal, improving eyesight, eliminating the phlegm.Cure mainly：Main difficult urination, stranguria with turbid discharge under band, oedema is swollen Full, summer-heat and damp rush down dysentery, hot eyes is hidden from view, the hot cough and asthma of phlegm.Chinese herbaceous peony flooding agent to various pathogens, such as staphylococcus aureus, dysentery in Song Shigella, Escherichia coli, Pseudomonas aeruginosa and typhoid bacillus also have different degrees of inhibitory action.
Polygonum perfoliatum：Classification systemPolygonum perfoliatum L., it is sour, it is cool, it is slightly poisonous.It is clearing heat and detoxicating, diuresis Detumescence, the medicine of analgesic.Cure mainly oedema, jaundice, diarrhea, malaria, pertussis, eczema, mange.This product decoction is to will he, Si Mici, good fortune The antibacterial potency of family name and bacterium sonnei is respectively 1:512,1:128,1:64,1:15.Additionally, this product decoction is to golden yellow Portugal Grape coccus, beta streptococcus, bacillus anthracis, corynebacterium diphtheriae, hay bacillus, typhoid bacillus, Pseudomonas aeruginosa and haemophilus influenzae Deng also there is stronger antibacterial action.
Milfoil：Scientific nameAchillea sibirca.Nature and flavor：It is bitter, pungent, mild-natured temperature, mild toxicity.The thoughts of returning home, liver and kidney channel.Effect：Dispel Wind analgesic, promoting blood circulation and stopping pain, it is clearing heat and detoxicating.Cure mainly sore Huang pyogenic infections, acute mastitis, abscess of throat, traumatic injury, venomous snake bite etc..10 % Fresh grass alcohol-soluble part flat board paper disk method, it is seen that to staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Fu Shi dysentery bars Bacterium has the inhibitory action of height.
Radix Glycyrrhizae：Classification systemGlycyrrhiza uralensis Fisch.It is sweet sweet, it is mild-natured and, enter 12 warps.It is raw to use It is cooler, can purging intense heat and detonicating, relieving spasm to stop pain；Process and use partial temperature, exterior cold, tonifying middle-Jiao and Qi can be dissipated.It is mainly used in weakness of the spleen and the stomach abdominal distension to receive It is slow-witted, it is weak, contraction pain in abdomen can be alleviated.Additionally, Radix Glycyrrhizae is also good at mediation property of medicine, emergency analgesic therapy solves the poison of hundred medicines, mitigates Toxic and side effect of the medicine to body or the stimulation to intestines and stomach, are neutralized by adding Radix Glycyrrhizae to play tonifying Qi, mediation each component Effect.
The principles of formulating prescriptions：The medicine typing and opinion suffered from diarrhoea are controlled according to Chinese veterinarian, grice diarrhoea is mostly being invaded by exogenous pathogen,invasion of exogenous pathogen, diet Hindered, internal organs void declines and functional disturbance causes, its external cause is the most obvious with the relation of wetness hyperactivity, and internal cause is relevant with taste.Therefore in Animal doctor with warming spleen and stomach for dispelling cold, help that spleen stomach invigorating, clearing heat and detoxicating, excreting dampness be astringent or styptic treatment for spontaneous sweating, gas wide, QI invigorating antidiarrheal, reinforcing spleen to promote digestion are treatment original in row Then.Wherein kuh-seng, garden burnet, the coptis, golden cypress heat-clearing and damp-drying drug, dispersing superficial exopathogens, purging intense heat convergence, assistant is to detoxify, anti-inflammatory；Chinese gall, Pu It is public English, wolf's milk, purslane clearing away heat to cure dysentery, excreting dampness antidiarrheal, clearing heat and detoxicating；Cordate houttuynia, hairyvein agrimony, Asiatic plantain, polygonum perfoliatum, milfoil The puckery property held back, contribute to eliminating dampness and heat, relieving diarrhea with astringents, reinforcing spleen to promote digestion, help with clearing heat and detoxicating, anti-inflammation and qi-restoratives；Radix Glycyrrhizae With purging intense heat and detonicating, relieving spasm to stop pain, tonifying middle-Jiao and Qi, the effect of coordinating the drug actions of a prescription.Complete square compatibility 5 is rigorous, can anti-inflammation, heat-clearing stop Rush down, strengthening the spleen and replenishing qi and stomach help digestion, strengthen the immunity of delactational piglets, improve cure rate, have no drug resistance, effectively control piglet abdomen Rush down the generation of disease.
The present invention is coupled by by yolk antibody IgY, probiotics and traditional Chinese medicine compound extract, is combined, and prepares association Same-action is in bacillary and viral grice diarrhoea pathogeny（Including bacterium and virus）High bioavilability targeted therapy for animals Medicine, traditional Chinese medicine compound extract is favorably improved medicine antibacterial bacteriostasis, improves diarrhoea pigling immunity and resistance against diseases, benefit The nutritional ingredient of raw saccharomycete and powdery yolk can stimulating immune system defence capability, reduce the generation of enteral noxious material, it is right Body's immunity has facilitation, can have certain development and application values as the feed addictive of high-quality.The present invention is carried A kind of preparation method for preventing and treating young ablactation diarrhea of pigs pharmaceutical composition for supplying, selection is convenient, extraction efficiency is high, cost is relative It is relatively low, it is easy to form large-scale production.
Specific embodiment
Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method, unless otherwise specified, is conventional method.Test material used in following embodiments, unless otherwise specified, is city Sell.Various Chinese medicine material medicines used in the present invention all can from a medical store or Reagent Company is commercially available, and its specification meets state Family or professional standard.
The specificity of embodiment 1 is anti-E.Coli- IgY, anti-TGE-PED-PoRV-IgY Yolk antibodies and corresponding powdery yolk Prepare
（1）Prepare the immunizing antigen and immunization protocol of variety classes high immunogenicity
1. the immunogene is that coli somatic proteantigen is immunized thing
Choose Escherichia coli（E.coli, O157:K88, CVCC1489, in being managed purchased from Chinese veterinary microorganism culture presevation The heart）.The culture of Main Pathogenic Bacteria is preferably is passed on blood agar slant medium, 37 DEG C culture 24 h, dyeing microscopic examination without Miscellaneous bacteria, obtains purebred.Main Pathogenic Bacteria is inoculated in LB broth bouillons（Include the grape of 5-10 % calf serums and 1-2 % Sugar）In be enlarged culture, put in 37 DEG C of constant-temperature shaking incubators (150-200 r/min), cultivate 24 h, be collected by centrifugation thin Bacterium cell.
The preparation of the immune thing of concentration proteantigen：Bacterium precipitation is added the PBS of 3 mL pH 7.0 according to every gram of wet bacterium The ratio of buffer solution, rinses 2-3 times.5 mL cell pyrolysis liquids are added by every gram of wet bacterium（Trade name：Bacterial cellular protein is cracked Liquid, purchased from Shanghai Sheng Gong bioengineering limited company）Ratio, suspension bacterial cell, 20-37 DEG C incubation 10-20 Min, while being shaken gently for.Homogenate ultrasonic grind instrument ultrasonication after bacterial cell is broken（Ultrasonic time 5s, Have a rest the s of time 10, ultrasonic 20 min, the W of power 400）, the min of 10000 rpm refrigerated centrifuges 10, discard bacterial debris precipitation, Supernatant is the crude extract containing destination protein.Take supernatant, the min of 18300 rpm refrigerated centrifuges 30.Supernatant is abandoned, precipitation uses 0.5 % sarcosyls suspend, 4 DEG C of effects 1 h, the min of 18300 rpm refrigerated centrifuges 30.Supernatant is abandoned, precipitation is steamed with double Water is suspended, and purpose mycoprotein concentration is determined with improvement Bradford methods, by its concentration regulation to 1.0 mg/mL.
The mycoprotein antigen for preparing is mixed with isometric Freund's complete adjuvant, it is repeatedly newborn using ultrasonic grind instrument Change 3-4 times（Emulsification 10-15 s, put 4 DEG C of refrigerators every time, are spaced 1 min）, until antigen and adjuvant mixture are emulsified completely, Antigen immune thing is obtained.
Immunization protocol：Alternate immune programme for children is formulated repeatedly, there is height to exempt from the healthy laying hen (180 of responsibility for selection Age in days), using muscle, subcutaneous multi-point injection formula approach, immunization interval is set as 10, is immunized 3 times（Fundamental immunity 1 time, reinforcement It is immune 2 times）, the immunization protocol that immunizing dosage is 0.50 mg.After head exempts from 1 week, the egg that the immune chicken of collection gives birth to is sample, Antibody titer is determined using ELISA method, when yolk antibody IgY antibody titer is more than 1:2000-3000, i.e. height exempt from egg, standby With.
2. the immunogene is transmissible gastroenteritis-pig epidemic diarrhea-porcine rotavirus
Using transmissible gastroenteritis of swine（TGE）- pig epidemic diarrhea（PED）- porcine rotavirus（PoRV）（G5 types）Three work epidemic diseases Seedling, is produced, veterinary drug new word by Wei Ke biotechnologys development company of Harbin Veterinary Medicine Inst., China Academy of Agriculture（2015） 080011097。
Immunization protocol：Live vaccine is diluted 0.5 mL/ part with SPSS, using intramuscular injection path. Select the healthy high-yield laying hen of 180 ages in days, first immunisation, the mL of live vaccine intramuscular injection 0.5；10 d are spaced, live vaccine is carried out the 2nd time Booster immunization, the mL of intramuscular injection 1；20 d are spaced, the 3rd booster immunization, the mL of intramuscular injection 1 are carried out using live vaccine.Adopted after being immunized for 3rd time Antibody detection is carried out with agar diffusion method, serum antibody titer is more than 1:When 32, collect height and exempt from egg, it is standby.
（2）Special yolk antibody IgY is separated using water dilution method-freezing
Height is exempted from into the egg tincture of iodine and cotton ball soaked in alcohol carries out disinfection treatment, separate yolk and egg white, puncture membrane of yolk, collected without film Egg yolk liquid and metered volume.Yolk liquid+deionized water is taken with 1:：Ratio mixing (v/v, the yolk liquid of each egg of 6-10 Averagely based on 15 mL), pH values are adjusted to 5.0-6.0, and -20 DEG C are overnight（About 8-10 h）.4 DEG C of defrostings, in the degreasing of precipitation Clear liquid and yolk precipitation carry out high speed refrigerated centrifuge respectively, and it is thick that the supernatant of merging is obtained special yolk antibody IgY Extract, the yolk sediment of merging is the nutritional ingredients such as lecithin.
By special yolk antibody IgY crude extracts（Supernatant）Freeze-drying is carried out, solid-state is respectively obtained and is resistedE.Coli- IgY Yolk antibodies dry powder and anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder, it is standby.Yolk sediment is sprayed respectively dry It is dry, prepare anti-E.ColiThe solid-state powdery yolk in-IgY antibody source and the solid-state powdery yolk in anti-TGE-PED-PoRV-IgY antibody source, It is standby.
The preparation of the probiotics yeast bacterium dry powder of embodiment 2, step is as follows：
（1）The preparation of culture medium：
Fluid nutrient medium：Composition includes the gL of glucose 50-1, the gL of yeast extract 2.0-1, brewer's wort 120 mL/L, MgSO4 0.5 g·L-1、FeSO4 0.1 g·L-1, the gL of urea 1.0-1.PH is to 6.0 for adjustment, 121 DEG C, and sterilize 30 min, packing to cone Shape bottle, it is standby.
Inclined-plane and plating medium（g·L-1）：Composition includes the gL of glucose 50-1, the gL of yeast extract 2.0-1, wheat Bud juice 120 mL/L, MgSO4 0.5 g·L-1、FeSO4 0.1 g·L-1, the gL of urea 1.0-1, the gL of agar 20-1.Adjustment To 6.0,121 DEG C, sterilize pH 20 min, and pour plate is standby.
（2）Saccharomycete culture：
Saccharomyces cerevisiae（Saccharomyces cerevisiae, purchased from Chinese veterinary microorganism culture presevation administrative center, to raise Material yeast, strain number 2.1011）, strain is chosen into a ring from inclined-plane storage kind and grows more plentiful saccharomycete lawn, connect In the fluid nutrient medium of 50 mL, 30 DEG C of incubated, about 24 h or so, you can be sent to logarithmic phase, now shake up bacterium solution are put, It is transferred in the fluid nutrient medium of 100 mL, 24 h is cultivated under the same terms, is obtained final product rejuvenation yeast.
Switching, Amplification Culture：Rejuvenation yeast is accessed into fluid nutrient medium, 30 DEG C, 40 r/min shaking table cultures, observation.Bacterium The inoculum concentration control for expanding Secondary Culture is planted in 6-10 %, less than 60 % to ensure dissolved oxygen amount, 30 DEG C of cultivation temperature turns liquid amount Fast 40 r/min constant-temperature tables cultures are increasing throughput, the pH value 6.0 of fluid nutrient medium, the % of sugared concentration 5, yeast extract concentration 2.0 g·L-1.Control bacterial content is with 9.0 × 108CFU/mL is advisable（3.0 Maxwell opacity tube concentration）.
（3）Prepare the lyophilized bacterium powder of saccharomycete：
Strain centrifugal concentrating（4 DEG C, under the conditions of 5000 rpm, 8-10 min are centrifuged）, collect wet thallus.Add respectively in wet thallus Enter basic freeze drying protectant and protein stabiliser（Below it is mass percent：3-5 % glycerine, 5-10 % skimmed milk powers, 10- 15% sucrose or mannitol, the beta-schardinger dextrin or Arabic gum of 10-15 %）, mix, the pH value of bacteria suspension is adjusted to 5.2 or so, Equilibration time is 60 min.- 20 DEG C of h of pre-freeze 12, the h of vacuum freeze drying 24, obtain probiotics yeast bacterium dry powder, and bacterium contains for control Amount is with 1.0-2.0 × 109CFU/g is advisable, vacuum sealing storage, standby.
The preparation method of the traditional Chinese medicine compound extract dry powder of embodiment 3, wherein the proportioning of each single medicinal material is as follows：
Chinese medicine composition is grouped into by each group of following weight portion：
Kuh-seng 10-30 parts, garden burnet 10-30 parts, coptis 10-20 parts, golden cypress 10-20 parts, Chinese gall 10-20 parts, dandelion 10-20 Part, wolf's milk 10-20 parts, cordate houttuynia 5-15 parts, hairyvein agrimony 5-15 parts, purslane 5-15 parts, Asiatic plantain 5-15 parts, polygonum perfoliatum 2- 15 parts, milfoil 2-15 parts, Radix Glycyrrhizae 5-15 parts.
The preparation method of traditional Chinese medicine compound extract dry powder is：
（1）Each single medicinal material is crushed through micronizer, 100-120 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus the 3-10 times of nothing of quality from Sub- water-cooled is soaked 30-60 minutes, is decocted 3 times, is decocted 1-3 hours every time, filter and remove residue, merges the filtrate for decocting, in 0.09MPa Under conditions of 50-70 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract dry powder.
Preferably, it is by step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 7-10 times The deionized water cold soaking of quality 30-60 minutes, decoct 2-3 hour, after filtering, then in filter residue addition the 3-6 times of nothing of quality from Sub- decocting is boiled twice, is decocted 1-2 hours every time, three liquids of decoction is merged, in 0.09MPa and 50-70 DEG C of condition Under, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract dry powder.
It is below several more excellent formulas and preparation method：
（1）The component proportion that traditional Chinese medicine compound extract 1（Unit：g）：Kuh-seng 15, garden burnet 15, the coptis 15, golden cypress 15, Chinese gall 15th, dandelion 15, wolf's milk 15, cordate houttuynia 10, hairyvein agrimony 10, purslane 10, Asiatic plantain 10, polygonum perfoliatum 8, milfoil 8, Radix Glycyrrhizae 10 Part.
The preparation method of No. 1 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 100 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 8 times of deionized waters of quality Cold soaking 40 minutes, decocts 2 h, decoction liquor filtering；Then 3 times of deionized waters of quality are added in filter residue, 1 h is decocted, is decocted Liquid is filtered；Filter residue adds 3 times of deionized waters of quality, decocts 1 h, decoction liquor filtering.Merge the decoction filtering of above-mentioned 3 times Liquid, under conditions of 0.09MPa and 60 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract and does Powder.
（2）The component proportion that traditional Chinese medicine compound extract 2（Unit：g）：Kuh-seng 10, garden burnet 10, the coptis 10, golden cypress 10, five Gall nut 10, dandelion 10, wolf's milk 10, cordate houttuynia 5, hairyvein agrimony 5, purslane 5, Asiatic plantain 5, polygonum perfoliatum 2, milfoil 2, Radix Glycyrrhizae 5.
The preparation method of No. 2 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 120 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 7 times of deionized waters of quality Cold soaking 30 minutes, decocts 2h, decoction liquor filtering；Then 3 times of deionized waters of quality are added in filter residue, 2 h, decoction liquor are decocted Filtering；Filter residue adds 3 times of deionized waters of quality, decocts 1 h, decoction liquor filtering.Merge the decoction filtering liquid of above-mentioned 3 times, Under conditions of 0.09MPa and 50 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract dry powder.
（3）The component proportion that traditional Chinese medicine compound extract 3（Unit：g）：Kuh-seng 30, garden burnet 30, the coptis 20, golden cypress 20, five Gall nut 20, dandelion 20, wolf's milk 20, cordate houttuynia 15, hairyvein agrimony 15, purslane 15, Asiatic plantain 15, polygonum perfoliatum 15, milfoil 15, Radix Glycyrrhizae 15.
The preparation method of No. 3 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 110 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 9 times of deionized waters of quality Cold soaking 60 minutes, decocts 3h, decoction liquor filtering；Then 5 times of deionized waters of quality are added in filter residue, 1.5 h are decocted, is decocted Liquid is filtered；Filter residue adds 4 times of deionized waters of quality, decocts 1.5h, decoction liquor filtering.Merge the decoction filtering of above-mentioned 3 times Liquid, under conditions of 0.09MPa and 55 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract and does Powder.
（4）The component proportion that traditional Chinese medicine compound extract 4（Unit：g）：Kuh-seng 20, garden burnet 20, the coptis 15, golden cypress 15, five Gall nut 15, dandelion 15, wolf's milk 15, cordate houttuynia 15, hairyvein agrimony 15, purslane 15, Asiatic plantain 15, polygonum perfoliatum 10, milfoil 10, Radix Glycyrrhizae 10.
The preparation method of No. 4 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 110 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 9 times of deionized waters of quality Cold soaking 50 minutes, decocts 3h, decoction liquor filtering；Then 4 times of deionized waters of quality are added in filter residue, 2 h, decoction liquor are decocted Filtering；Filter residue adds 3 times of deionized waters of quality, decocts 2h, decoction liquor filtering.Merge the decoction filtering liquid of above-mentioned 3 times, Under conditions of 0.09MPa and 65 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract dry powder.
（5）The component proportion that traditional Chinese medicine compound extract 5（Unit：g）：Kuh-seng 25, garden burnet 25, the coptis 15, golden cypress 15, five It is gall nut 10, dandelion 10, wolf's milk 10, cordate houttuynia 10, hairyvein agrimony 10, purslane 10, Asiatic plantain 10, polygonum perfoliatum 5, milfoil 5, sweet Grass 5.
The preparation method of No. 5 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 100 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 10 times of quality without ion Water-cooled is soaked 60 minutes, decocts 2h, decoction liquor filtering；Then 6 times of deionized waters of quality are added in filter residue, 1 h is decocted, is decocted Liquid is filtered；Filter residue adds 4 times of deionized waters of quality, decocts 1h, decoction liquor filtering.Merge the decoction filtering of above-mentioned 3 times Liquid, under conditions of 0.09MPa and 70 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract and does Powder.
（6）The component proportion that traditional Chinese medicine compound extract 6（Unit：g）：Kuh-seng 15, garden burnet 15, the coptis 12, golden cypress 12, five It is gall nut 12, dandelion 15, wolf's milk 10, cordate houttuynia 10, hairyvein agrimony 12, purslane 12, Asiatic plantain 10, polygonum perfoliatum 8, milfoil 6, sweet Grass 6.
The preparation method of No. 6 dry powder of traditional Chinese medicine compound extract is：
（1）Each single medicinal material is crushed through micronizer, 120 mesh sieves are crossed；
（2）By step（1）In each Ultramicro-powder single medicinal material for obtaining by formula ratio prescription, mixing, plus 10 times of quality without ion Water-cooled is soaked 60 minutes, decocts 2h, decoction liquor filtering；Then 5 times of deionized waters of quality are added in filter residue, 1 h is decocted, is decocted Liquid is filtered；Filter residue adds 5 times of deionized waters of quality, decocts 1h, decoction liquor filtering.Merge the decoction filtering of above-mentioned 3 times Liquid, under conditions of 0.09MPa and 70 DEG C, vacuum-concentrcted, concentrate spray drying obtains final product traditional Chinese medicine compound extract and does Powder.
The preparation method of the medicinal composition powders of embodiment 4, step is as follows：
Pharmaceutical composition of the invention includes following each component：
It is anti-E.Coli- IgY Yolk antibody dry powder 2.5-10wt %, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder 2.5-10 Wt %, traditional Chinese medicine compound extract dry powder 60-90 wt %, powdery yolk 5-30 wt %, probiotics yeast bacterium dry powder 0.5 × 108--5.0 ×108CFU/g；Wherein, resistE.Coli- IgY Yolk antibodies dry powder, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder, Chinese medicine The mass percent of compound extract dry powder and powdery yolk and be 100%.
Wherein, resistE.Coli- IgY Yolk antibodies, anti-TGE-PED-PoRV-IgY Yolk antibodies and powdery yolk are according to implementation It is prepared by the method in example 1.
Powdery yolk is by resistingE.ColiThe solid-state powdery yolk in-IgY antibody source and consolidating for anti-TGE-PED-PoRV-IgY antibody source State powdery yolk is constituted, and both mass ratioes are 1:1.
Traditional Chinese medicine compound extract dry powder is prepared according to the method in embodiment 3.
Probiotics yeast bacterium dry powder is prepared according to the method in embodiment 2.
The preparation method of pharmaceutical composition of the invention is：
First will be anti-E.Coli- IgY Yolk antibodies dry powder carries out waiting weight portion with anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder Mixing, then mixed Yolk antibody dry powder is mixed with traditional Chinese medicine compound extract dry powder and powdery yolk according to formula ratio, and add Enter probiotics yeast bacterium dry powder（Addition quantity is 0.5 × 108--5.0×108CFU/g）Mixed, that is, be made pharmaceutical composition Powder.
It is below several more excellent formulas and preparation method：
（1）Pharmaceutical composition 1：It is prepared from by following each component：
It is anti-E.Coli- IgY Yolk antibodies dry powder 10 wt%, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 10, Chinese medicine Compound extract No. 1 wt% of dry powder 70, the wt% of powdery yolk 10；Probiotics yeast bacterium dry powder 5.0 × 108 CFU/g。
（2）Pharmaceutical composition 2：It is prepared from by following each component：
It is anti-E.Coli- IgY Yolk antibodies dry powder 2.5 wt%, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 2.5, in Medicine compound extract No. 2 wt% of dry powder 65, the wt% of powdery yolk 30；Probiotics yeast bacterium dry powder 2.0 × 108 CFU/g。
（3）Pharmaceutical composition 3：It is prepared from by following each component：
It is anti-E.ColiThe wt% of-IgY Yolk antibodies dry powder 5, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 5, Chinese medicine is multiple Square extract solution No. 3 wt% of dry powder 85, the wt% of powdery yolk 5；Probiotics yeast bacterium dry powder 1.0 × 108 CFU/g。
（4）Pharmaceutical composition 4：It is prepared from by following each component：
It is anti-E.ColiThe wt% of-IgY Yolk antibodies dry powder 6, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 6, Chinese medicine is multiple Square extract solution No. 4 wt% of dry powder 73, the wt% of powdery yolk 15, probiotics yeast bacterium dry powder 3.0 × 108 CFU/g。
（5）Pharmaceutical composition 5：It is prepared from by following each component：
It is anti-E.ColiThe wt% of-IgY Yolk antibodies dry powder 8, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 8, Chinese medicine is multiple Square extract solution No. 5 wt% of dry powder 64, the wt% of powdery yolk 20, probiotics yeast bacterium dry powder 4.0 × 108 CFU/g。
（6）Pharmaceutical composition 6：It is prepared from by following each component：
It is anti-E.ColiThe wt% of-IgY Yolk antibodies dry powder 5, the anti-wt% of TGE-PED-PoRV-IgY Yolk antibodies dry powder 5, Chinese medicine is multiple Square extract solution No. 6 wt% of dry powder 70, the wt% of powdery yolk 20, probiotics yeast bacterium dry powder 5.0 × 108 CFU/g。
The preparation method of above-mentioned more excellent formula is：
First will be anti-E.Coli- IgY Yolk antibodies dry powder carries out waiting weight portion with anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder Mixing, then mixed Yolk antibody dry powder is mixed with traditional Chinese medicine compound extract dry powder and powdery yolk according to formula ratio, and add The probiotics yeast bacterium dry powder for entering formula ratio is mixed, that is, be made medicinal composition powders.
When aforementioned pharmaceutical compositions powder is used as feed addictive, medicinal composition powders are in weanling pig basal diet In addition be 10--30 %（W/v, is mass/volume percentage when drinking-water is taken.W/w, feed is mixed clothes and is added as feed Plus during agent be mass/mass percentage）.
Pharmaceutical composition of the invention can be applied in grice diarrhoea disease is prevented and treated, and be particularly useful for preparing and prevent and treat disconnected The medicine of milk grice diarrhoea, when using, can be by medicinal composition powders spice mixed feeding, or by digesting canal drug administration after drinking-water.Make Use method：Drinking water administration or feed mix clothes administration.Consumption：200 g/L（W/v, drinking-water）, 200 g/kg（W/w, feed mix clothes and Feed addictive）, 2 times/d, 5 d are 1 course for the treatment of.
The performance test research of the pharmaceutical composition of the invention of embodiment 5
Through experiment, the pharmaceutical composition prepared in the method for the embodiment of the present invention 4 is respectively provided with good stability, safety Property, nontoxicity.The traditional Chinese medicine compound extract prepared in the method for the embodiment of the present invention 3 is respectively provided with good In Vitro Bacteriostasis Can, can be good at suppressing EHEC, Salmonella paratyphi B, proteus, Pseudomonas aeruginosa, Staphylococcus aureus Bacterium, micrococcus scarlatinae, streptococcus dysgalactiae, bacillus subtilis etc., wherein with the bacteriostasis property of traditional Chinese medicine compound extract 6 most It is good；In clinical effect trial, the pharmaceutical composition that the method for the embodiment of the present invention 4 is prepared has to diarrhea of weaned piglets There is good curative effect, wherein optimal with the curative effect of pharmaceutical composition 6.Traditional Chinese medicine compound extract given below 6, drug regimen The effect test that thing 6.
Test the bacteriostatic test of 1 traditional Chinese medicine compound extract dry powder of the invention
1 materials and methods
The MIC value of Chinese medicine is determined using test tube doubling dilution joint agar plate method, by cylinder-plate method（Filled using Oxford cup The In Vitro Bacteriostasis method of liquid）The bacteriostatic activity intensity of medicine is observed respectively.Dilution is MH meat soups, and Spawn incubation uses brother's human relations Than sub- blood plate.
Test medicine：No. 1 dry powder of traditional Chinese medicine compound extract, No. 5 dry powder and No. 6 dry powder, according to method system shown in embodiment 3 It is standby.
No. 1 dry powder of Chinese medicine extract, No. 5 dry powder and No. 6 dry powder are chosen, appropriate amount of drug dry powder is weighed, and matched somebody with somebody with purified water Suspension is made, No. 1, No. 5 and No. 6 medicine stoste is prepared respectively（Concentration is 500 mg/mL）, in 121 DEG C of autoclavings 15 Min, puts 4 DEG C of Refrigerator stores, standby.
Comparison medicine：Dual coptis powder, No. 2 TCM Factory's production, Chinese medicines quasi-word Z10940044, specification 600 Mg/ branch, medicine stoste compound concentration is 600 mg/mL.
Test strain：EHEC（E. coli) [CMCC (B) 44102], Salmonella paratyphi B （Salmonella paratyphi B）[CMCC（B）50094], proteus (Proteusbacillus vulgaris) [CMCC (B) 49027], Pseudomonas aeruginosa (Pseudomonas aeruginosa) [CICC10419], staphylococcus aureus （S. aureus) [CMCC (B) 26003], micrococcus scarlatinae (S. pyogenes) [ATCC19615], streptococcus dysgalactiae (S. dysgalactiae) [ATCC 35666], bacillus subtilis (Bacillus subtilis) [CMCC(B) 63501], respectively purchased from China General Microbiological culture presevation administrative center and Chinese industrial Microbiological Culture Collection and management The heart, with ATCC official authorizations.
1.1 extracorporeal bacteria inhibitor tests
1.1.1 test tube doubling dilution combines agar plate method
The preparation of quantity of microorganism inoculated：Take each bacterium（Bacterium）Broth culture, it is molten with 0.9% SPSS by 10 times of dilution methods Liquid, prepares concentration equivalent to 0.5 Maxwell standard opacity tube for examination bacteria suspension, then with MH broth dilutions to bacteria containing amount for 1 × 107CFU/mL, it is standby.
The step of test tube method dilution medicine stoste：Test group is identical with the dilution process of control group.Take 22 5 mL aseptic Centrifuge tube, test group 11, control group 11, numbering.Every group except 1# manage in addition to, remaining each pipe（2# pipes to 11# manage）Add 2 The MH meat soups of mL（For bacterium, streptococcus is using the MH meat soups for adding 2 % calf serums）, in 1# pipes, 2# pipes respectively 2 mL Bactericidal medicine stostes are added, then 1# pipes are antibacterials stoste.The medicine liquid and nutrient broth of 2# pipes are mixed, is suctioned out 2 mL add 3# pipes, the like, doubling dilution to 10# is managed, and suctioning out 2 mL from 10# pipes discards, and the 11# pipes of test group are The not growth control of drug containing.The 1# pipes of control group only add 2 mL liquids, are not added with MH meat soups and bacterium, are liquid control.It is right 11# pipes according to group only add 2 mL MH meat soups, are not added with liquid and bacterium, are blank.
Inoculation test group 1# pipes to 11# manage, often pipe sequentially add 100 μ L dilution bacterium solution, the branch pipe of control group 11, no Plus the bacterium solution of dilution.It is placed in and 18-24 h is incubated in 37 DEG C of constant incubators.
As a result preliminary judgement：Each pipe of each pipe of test group drug concentration identical with control group respectively is contrasted.If test tube Internal haze, has illustrated bacterial growth；If clear in test tube, illustrates without bacterial growth.Contain liquor strength without bacterial growth The lowest, as this kind medicine is to various bacteria and the minimum inhibitory concentration MIC of fungi.
To blood plate be placed in for the culture of Chinese medicine and bacterium in test group and control group respectively pipe, streak inoculation by transferred species 18-24 h are incubated in 37 DEG C of constant incubators.
Bacterial growth is whether there is on result judgement observation blood plate, the corresponding test tube without bacterial growth on blood plate, as Minimum inhibitory concentration of the test medicine to the bacterium（MIC）.Each bacterium is repeated 3 times, and averages.
1.1.2 cylinder-plate method：
The preparation of culture medium flat plate：Caseinhydrolysate agar medium（pH 7.2-7.4）Prepared by catalogue.121 DEG C sterilizing 15 min, treat that temperature is down to 50~60 DEG C or so, pour into sterilized petri dishes, make agar thickness be 4 mm.Streptococcus uses Containing the de- fiber Sheep Bloods of 5 %（v/v）MH agar plates.MH agar plates should put 35 DEG C of 30 min of incubation before, make its table Face dries.
Oxford cup fills liquid method：The bacterium solution sterile saline of preparation is diluted to optimal coating concentration, with sterilizing Cotton swab dips bacterium solution, and unnecessary bacterium solution is extruded along tube wall, and then even spread is to MH agar plates, totally 3 times, each rotary flat 60 ° of plate, finally smears one week, along flat board inner edge to obtain uniform inoculation.Flat board is added a cover and is placed in superclean bench, is dried extremely Few 3 min, no more than 15 min.The Oxford cup that will sterilize is micro- on alcolhol burner to burn, equidistant in each agar plate surface to put 4 Oxford cups are put, and the μ L of tested liquid 200 are added in every Oxford cup（Drug concentration chooses 500 mg/mL）.In ox 2 sterilizing filter papers are covered on the cup of Tianjin（In case the globule falls down influence experimental result during culture）, closing lid, plate horizontal, in 37 DEG C of perseverances 20~24 h are incubated in warm incubator.
Result judgement：Inhibition zone diameter is less than 10 mm, is without bacteriostasis；It is bacteriostasis between 10~15 mm It is weaker；It is moderate strength bacteriostasis between 15~20 mm；It is with stronger bacteriostasis more than 20 mm.
2 results and analysis
Primarily determined that by preliminary experiment, Chinese medicine extract 1, No. 5, No. 6 dry powder be respectively provided with good fungistatic effect, traditional Chinese medicine extraction The medicine stoste of No. 6 dry powder of liquid has most strong fungistatic effect compared with No. 1 dry powder, No. 5 dry powder, therefore Chinese medicine extract is chosen in formal test The medicine stoste of No. 6 dry powder is tested.Result of the test is shown in Table 1.
The minimal inhibitory concentration (MIC) and antibacterial activity of 1 Chinese medicine extract dry powder of table 6
3 conclusions
No. 6 dry powder of Chinese medical extract have bacteriostasis to livestock and poultry common pathogen, wherein to streptococcus and golden yellow grape The fungistatic effect of coccus is stronger, and MIC value also has preferable between 15.6-31.25 mg/mL to general pathogenic enterobacteria Bacteriostasis, MIC value has moderate strength bacteriostasis in 31.25-62.5 mg/mL or so to strong pathogenic enterobacteria.Compared with Swap buffers medicine, both are more or less the same at MIC value, are respectively provided with middle intensity bacteriostasis to encountered pathogenic bacteria, and fungistatic effect is relatively managed Think.
Cylinder-plate method result is consistent with MIC value, in the case of the mg/mL of drug concentration 500, No. 6 dry powder of Chinese medical extract There is preferable fungistatic effect and bactericidal action to streptococcus and staphylococcus aureus, inhibition zone diameter is between 22.0- 26.0 mm scopes；There is the bacteriostatic activity of moderate strength to common enterobacteria, inhibition zone diameter between 16.0-21.0 mm it Between.Than swap buffers medicine, the antibacterial circle diameter and fungistatic effect of No. 6 dry powder of Chinese medical extract are better than comparison medicine, therefore to intestines Road bacterial diarrhea has therapeutic action.
Test the acute toxicity test of 2 pharmaceutical compositions of the invention
1 materials and methods
Test drug：Medicinal composition powders, prepare according to the method for the embodiment of the present invention 4, and Chinese medicinal compound extract therein is No. 6 dry powder of traditional Chinese medicine compound extract, the formula of pharmaceutical composition is pharmaceutical composition 6.
Experimental animal：Kunming system cleaning grade mouse 60, the g-22 g of Mouse Weight 18, male and female half and half, female mice is not produced Unpregnancy.Rearing conditions：Experiment cleaning grade particles mouse grain（Soviet Union raises and examines（2008）01008）, 22 ± 2 DEG C of indoor temperature, relative humidity It is 30 %~60 %, free choice feeding and drinking-water.Continuous Observation 1 week, clinical health is that can be used to test, and is routinely raised during experiment Support.
2 acute toxicity tests
Healthy mice is randomly divided into dose ratio between 5 dosage groups and 1 control group, group and is set to 1:1.2.Every group 10, male and female Half and half, and every mouse is weighed.4 h fasting after 8 h, gavage before gavage, but can free water.
Each drug dose group according to mouse weight, according to the medicine agent of 1.20,1.44,1.72,2.07,2.49 g/kg Amount calculates dosage, and administration is orally fed with isoconcentration oral liquid, and daily maximum gavage amount is no more than 1.0 mL/ only.Control group Gavage physiological saline.All mouse free choice feedings and drinking-water during experiment.
Observation index：After administration in 3 h, 1 health status of mouse, poisoning manifestations and dead mistake are observed every 30 min Journey, including feeding drinking-water, behavior and reaction, autonomic activities, be short of breath or slow down state, by hair shatter value, symptom starting when Between and the duration, it is dead situations such as；Afterwards, observed 1 time every 2 h；After 2 d, observation 6 times daily, the d of Continuous Observation 7, system The meter death rate.
3 results and analysis
After administration gavage, middle and high dosage group mouse（1.72nd, 2.07,2.49 g/kg dosage groups）There is spiritual depressed, activity The phenomenon of reduction, has no other obvious abnormal responses.2nd d after gavage, 1.20,1.44,1.72 g/kg dosage groups mouse drink Food, amount of drinking water have a certain degree of increase, and activity increases, and the state of mind is good, abnormal response does not occur.3-7 after gavage D, the 2.07 g/kg dosage group mouse state of mind are gradually recovered, and appetite is good, toxic reaction do not occur, and 2.49 g/kg medicines Dosage group mouse, spirit is not good enough, 1 dead mouse occurs to off-test, and dead mouse cut open inspection inspection result shows, internal organs without Substantive ANOMALOUS VARIATIONS, coat of the stomach has black region, has no obvious bleeding, extravasated blood.
4 conclusions
Result shows, using the LD of improvement karber's method calculating pharmaceutical composition50It is 3637.10 mg/kg, 95 % confidential intervals For：3248.6--4135.6 mg/kg.According to《Veterinary drug test requirement specification collects》Middle acute toxicity test regulation, in external source In learning thing acute toxicity grading criteria, mouse once oral LD50In the range of 500~5000 mg/kg, you can think the medicine Thing is substantially nontoxic.
Test the observation of the immune protection effect and production performance of 3 pharmaceutical composition diarrhoea piglets of the invention
1 materials and methods
1.1 test materials
Test drug：Medicinal composition powders, prepare according to the method for the embodiment of the present invention 4, and Chinese medicinal compound extract therein is No. 6 dry powder of traditional Chinese medicine compound extract, the formula of pharmaceutical composition is pharmaceutical composition 6.
Experimental animal：Lanzhou Yuzhong Peace Town Lanzhou kind pig farm new life delactational piglets.
1.2 test methods
1.2.1 experimental animal and packet
Ternary [Du × (big × long)] hybridization piglet 36 of 28 ages in days wean is chosen in experiment, and average original body mass is 5.22 ± 0.89 kg, using 2 treatment 3 repeat two factor designs, 2 treatment be respectively medicine group and control group, 3 repetitions of each treatment group, Each repeats 6 piglets.
1.2.2 feeding and management
All equal overbit piercing marks of piglet of participating in the experiment, each repeats feeding piglet in a column, and free choice feeding, free water are immune to disappear Malicious program is carried out by pig farm conventional method.The age in days of piglet 21 rises observes its clinical symptoms, and clinical symptoms person without exception is in 28 ages in days Start experiment.Consumption and usage：Control group fed basal diet, medicine group presses 200 g/kg dosage on this basis, and spice is mixed Raise, 2 times/d, be used in conjunction 3 d, Continuous Observation 14 days.
1.3 Observe and measure indexs
The spirit of daily observation pig, feeding, drinking-water, excrement and active situation, diarrhoea degree with the time started of suffering from diarrhoea, persistently when Between and Scours index react.Record each repeat piglet daily always suffer from diarrhoea head number, and individually record diarrhoea be in water sample head number, system Meter Incidence of Diarrhea and severe diarrhea incidence.On-test and terminate before carry out weekly individuality weigh, ramming material, with each repetition For unit calculates average daily gain.Record consumption flow vector, average feed-weight ratio is calculated in units of each repetition.
2 results and analysis
2.1 pairs of immune protection effects of delactational piglets diarrhoea（It is shown in Table 2）
Influence of the different disposal group of table 2 to diarrhea of weaned piglets rate
2.2 pairs of influences of delactational piglets production performance（It is shown in Table 3）
Influence of the different disposal group of table 3 to Production Performance of Weaning Pigs
Note：Data use average ± standard error in table（X±SE）Represent, compared with control group, * * represent that difference extremely shows Write,p＜ 0.01, * represents significant difference,p＜ 0.05.
3 conclusions
3.1 than control group, and the pharmaceutical composition can significantly reduce the diarrhea rate of weanling pig, severe diarrhea rate and death Rate, wherein diarrhea rate decline without serious watery diarrhea and the generation of sick pig death state in 27.80 percentage points, and 3 repeating groups, Therefore, pharmaceutical composition of the invention can improve the survival rate of delactational piglets, have to alleviating and preventing and treating delactational piglets diarrhoea Obvious clinical effectiveness.
3.2 pharmaceutical compositions significantly improve the daily gain of weanling pig（p＜ 0.01）, daily ingestion amount（p＜ 0.01）, Reduce feed-weight ratio（p＜ 0.05）, promote weanling pig growth, help to improve weaned piglets, it is disconnected with preferably raising The trend of suckling piglet growth rate and weightening of preferable stage advantage, there is good preventing and treating effect in terms of piglet digestive disease is prevented Really.
Test therapeutic test of 4 pharmaceutical compositions of the invention to clinical type diarrhoea piglet
1 materials and methods
1.1 test materials
Test drug：Medicinal composition powders, prepare according to the method for the embodiment of the present invention 4, and Chinese medicinal compound extract therein is No. 6 dry powder of traditional Chinese medicine compound extract, the formula of pharmaceutical composition is pharmaceutical composition 6.
Experimental animal：Big-and-middle-sized kind of Lanzhou periphery pig farm, individual pig farm, Yuzhong Peace Town Lanzhou kind pig farm ablactation abdomen Piglet is rushed down, indivedual piglets are in serious watery diarrhea symptom.
1.2 clinical diarrhoea piglet therapeutic tests
1.2.1 therapeutic scheme
Drug usage and dosage：Dosage is 200 g/kg, and spice mixed feeding is offerd medicine 2 times daily, takes medicine continuously 3 days, is within 5 days 1 treatment Journey, the d of Continuous Observation 10.
1.2.2 criterion is cured：Symptom of diarrhea stops and thereafter lets out excrement normally, and piglet appetite recovers normal, And the active situation such as the state of mind, drinking-water is normal.
2 observation of curative effect results
Treatment diarrhoea piglet 160, cures 38 on 2nd altogether, accounts for 23.75 %；Cure 77 within 3rd, account for 48.13 %；Cure within 4th 36, account for 22.50 %.9 severe diarrhea piglets（Account for 5.63 %）, after first course for the treatment of terminates, do not recover completely, to At the end of two courses for the treatment of, return to normal.Piglet of being suffered from diarrhoea during medication treatment does not produce toxic and side effect, the stream in later stage to medicine Row disease is learned curative effect follow-up study result and is shown, rear piglet occurs without diarrhoea recurrence.
There are no the piglet phenomena of mortality of suffering from diarrhoea during whole observation of curative effect.
3 conclusions and analysis
Pharmaceutical composition of the invention is made up of special yolk antibody, probiotics, Chinese medicinal compound extract and powdery yolk.It is clinical Its order of severity of common grice diarrhoea and Escherichia coli, the propagation of cause diarrhea virus and propagation degree direct proportionality.Examination Test result to show, the yolk antibody IgY in the pharmaceutical composition is orientable to kill disease-producing pathogens, reduces gradient of infection, prognosis Recurrence is difficult, illustrates that it also has good same group's immune protective effect.Meanwhile, Chinese medicine compound prescription active principle also has to pathogen There is good antibacterial killing action, and have the effect for improving pigling immunity and premunition concurrently.Probiotics yeast bacterium therein passes through The balance of microorganism species in regulating intestinal canal, can improve the non-specific anti-infection ability of piglet, and powdery yolk also has higher Nutritive value, has vital effect to effectively preventing and treating grice diarrhoea, raising delactational piglets survival rate and production performance.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, Although being described in detail to the present invention with reference to the foregoing embodiments, for a person skilled in the art, it still may be used Modified with to the technical scheme described in foregoing embodiments, or equivalent is carried out to which part technical characteristic. All any modification, equivalent substitution and improvements within the spirit and principles in the present invention, made etc., should be included in of the invention Within protection domain.

Claims (10)

1. a kind of pharmaceutical composition for preventing and treating diarrhea of weaned piglets, it is characterised in that：Described pharmaceutical composition is by following Component is constituted：It is anti-E.Coli- IgY Yolk antibodies dry powder, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder, Chinese medicine compound prescription are extracted Liquid dry powder, powdery yolk and probiotics yeast bacterium dry powder；
The traditional Chinese medicine compound extract dry powder is prepared from by each component of following mass fraction：Kuh-seng 10-30 parts, garden burnet 10- 30 parts, coptis 10-20 parts, golden cypress 10-20 parts, Chinese gall 10-20 parts, dandelion 10-20 parts, wolf's milk 10-20 parts, cordate houttuynia 5-15 parts, hairyvein agrimony 5-15 parts, purslane 5-15 parts, Asiatic plantain 5-15 parts, polygonum perfoliatum 2-15 parts, milfoil 2-15 parts, Radix Glycyrrhizae 5- 15 parts.
2. pharmaceutical composition according to claim 1, it is characterised in that：The traditional Chinese medicine compound extract dry powder is by following matter The each component for measuring number is prepared from：15 parts of kuh-seng, 15 parts of garden burnet, 12 parts of the coptis, 12 parts of golden cypress, 12 parts of Chinese gall, dandelion 15 parts, 10 parts of wolf's milk, 10 parts of cordate houttuynia, 12 parts of hairyvein agrimony, 12 parts of purslane, 10 parts of Asiatic plantain, 8 parts of polygonum perfoliatum, milfoil 6 Part, 6 parts of Radix Glycyrrhizae.
3. pharmaceutical composition according to claim 1, it is characterised in that：In described pharmaceutical composition, the prebiotic ferment The addition quantity of female bacterium dry powder is 0.5 × 108--5.0×108CFU/g；It is described anti-E.ColiIt is-IgY Yolk antibodies dry powder, anti- The mass percent sum of TGE-PED-PoRV-IgY Yolk antibodies dry powder, traditional Chinese medicine compound extract dry powder and powdery yolk is 100%, wherein, resistE.Coli- IgY Yolk antibody dry powder 2.5-10%, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder 2.5-10%, traditional Chinese medicine compound extract dry powder 60-90%, powdery yolk 5-30%；
Preferably, described anti-E.Coli- IgY Yolk antibodies dry powder is 5%, anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder It is 5%, traditional Chinese medicine compound extract dry powder is 70%, and powdery yolk is 20%.
4. pharmaceutical composition according to claim 1, it is characterised in that：The probiotics yeast bacterium dry powder is by saccharomyces cerevisiae It is prepared from, bacterial content is 1.0-2.0 × 109 CFU/g。
5. pharmaceutical composition according to claim 1, it is characterised in that：It is described anti-E.Coli- IgY Yolk antibody dry powder Preparation method is：Healthy laying hen is immunized using the immune thing of coli somatic proteantigen, egg is collected, using water Dilution method-freezing separates special yolk antibody IgY, and supernatant is anti-E.Coli- IgY Yolk antibodies, supernatant is cold It is lyophilized it is dry after, resistedE.Coli- IgY Yolk antibody dry powder.
6. pharmaceutical composition according to claim 1, it is characterised in that：The anti-TGE-PED-PoRV-IgY Yolk antibodies The preparation method of dry powder is：Transmissible gastroenteritis of swine-pig epidemic diarrhea-porcine rotavirus G5 types trigeminal live vaccine is immune strong Health laying hen, collects egg, and special yolk antibody IgY is separated using water dilution method-freezing, and supernatant is anti-TGE- PED-PoRV-IgY Yolk antibodies, by supernatant freeze-drying after, obtain anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder.
7. pharmaceutical composition according to claim 1, it is characterised in that：The powdery yolk is by resistingE.Coli- IgY antibody source Solid-state powdery yolk and the weight portion such as solid-state powdery yolk in anti-TGE-PED-PoRV-IgY antibody source mix；
It is described anti-E.ColiThe preparation method of the solid-state powdery yolk in-IgY antibody source is：Exempted from using coli somatic proteantigen Epidemic disease thing is immunized to healthy laying hen, collects egg, is separated using water dilution method-freezing, and the spraying of yolk sediment is dry It is dry, resistedE.ColiThe solid-state powdery yolk in-IgY antibody source；
The preparation method of the solid-state powdery yolk in the anti-TGE-PED-PoRV-IgY antibody source is：By transmissible gastroenteritis of swine-pig Epidemic diarrhea-porcine rotavirus G5 type trigeminal live vaccine immune health laying hens, collects egg, using water dilution method-freezing Method is separated, and by the spray drying of yolk sediment, obtains the solid-state powdery yolk in anti-TGE-PED-PoRV-IgY antibody source.
8. pharmaceutical composition according to claim 1, it is characterised in that：The preparation side of the traditional Chinese medicine compound extract dry powder Method is：After each component of formula ratio is crushed, plus 3-10 times of decocting is boiled 3 times, and each 1-3 hours, merging filtrate, vacuum decompression is dense Contracting, concentrate spray drying, obtains traditional Chinese medicine compound extract dry powder.
9. the preparation method of any described pharmaceutical compositions of claim 1-8, it is characterised in that：First will be anti-E.Coli- IgY ovum Yellow antibody dry powder is carried out waiting weight portion to mix with anti-TGE-PED-PoRV-IgY Yolk antibodies dry powder, then mixed yolk is resisted Soma powder mixes with traditional Chinese medicine compound extract dry powder and powdery yolk according to formula ratio, and adds probiotics yeast bacterium dry powder to be mixed Close, that is, be made pharmaceutical composition.
10. any described pharmaceutical compositions of claim 1-8 are preparing the medicine of preventing and treating diarrhea of weaned piglets or feed addition Application in agent.
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CN107712376A (en) * 2017-12-10 2018-02-23 阜阳市玉琪农业科技有限公司 A kind of pig starter feed of pre- anti-diarrhea and preparation method thereof
CN107961284A (en) * 2017-12-29 2018-04-27 四川农业大学 A kind of oral formulations for preventing grice diarrhoea and preparation method thereof
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