CN1682843A - Chinese medicine composition for preventing bird flu - Google Patents
Chinese medicine composition for preventing bird flu Download PDFInfo
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Abstract
The present invention relates to a kind of Chinese medicine composition for preventing bird flu of chicken and duck. The Chinese medicine composition is prepared with forsythia 90-110 weight portions, honeysuckle 40-60 weight portions, northern isatis root 25-45 weight portions, skullcap root 40-60 weight portions, basket fern rhizome 40-60 weight portions and astragalus root 70-90 weight portions. The Chinese medicine composition is for fowls to take orally, has obvious bird flu preventing effect and is safe and convenient.
Description
Technical field
The present invention relates to a kind of Chinese medicine for oral administration composition and method of making the same and application that is used to prevent poultry bird flus such as chicken, duck.
Background technology
Bird flu is a kind of acute height lethal infectious disease of the caused birds of A type bird flu virus.Poultry such as chicken, turkey, duck and Carnis Coturnicis japonicae and wild bird, aquatic bird, seabird etc. all can infect, but based on chicken and turkey.Multiple animal and human's classes such as pig, cat, monkey also can infect.
Bird flu occurred in Italy early than 1878, once was called as fowl plague or fowl plague in history.Over more than 100 year, bird flu is constantly being attacked the aviculture of All Around The World, and directly threatens human beings'health and life.1997, the bird flu of Hong Kong outburst caused poultry dead on a large scale, and successively has 18 people to infect bird flu virus, wherein 6 people's death; In December, 2003 to 2004 year March, the H5N1 type bird flu in the Southeast Asia outburst involved nearly all Southeast Asian countries and regions, cause the death of catching an illness of tens million of poultry, and the people infected bird flu virus and death surplus in the of 20.
Though bird flu is found the history in existing more than 100 year, human so far prevention in full force and effect, treatment and the control technology do not grasped as yet.Avian influenza vaccine is the main means of birds flu-preventing outburst.If the vaccine that uses is consistent with the hypotype of the virus of invasion and attack, then vaccine can reach more than 90% the protective rate of poultry.But isolated at present bird flu has 15 kinds of H antigens and 9 kinds of N antigens, two kinds of antigenic various combinations can form up to a hundred blood serum subtypes, and popular virus subtype instability, producing " drift ", this brings very big difficulty for the development of vaccine and the diagnosis and the control of bird flu.
Virus is the microorganism that parasitizes in host's living cells, all virus is had suppress or the medicine of killing action, host cell is also had certain toxicity, this brings very big difficulty for development of antiviral drugs.Chemotherapeutics at influenza and bird flu virus mainly is amantadine and rimantadine at present.Ribavirin is a kind of broad-spectrum antiviral medicament, also is usually used in treating influenza clinically, but viral inhibition test shows that ribavirin there is no inhibitory action external to influenza virus.Amantadine and rimantadine have better effects at the application initial stage to prevention chicken, duck bird flu, but general, prolonged application along with this class medicine, bird flu virus produces serious Drug resistance to this class medicine, causes this class medicine to use no positive effect clinically.The influenza new drug zanamivir (Zaramivir) and the Ao Simiwei (Oseltamivir) of listing in 1999 have definite prevention effect to human influenza, and the prevention of poultry bird flu is not appeared in the newspapers.But these two kinds of medicines cost an arm and a leg, even bird flu effectively also is difficult to use in aviculture.Therefore in the control of bird flu, also there is not ideal chemotherapeutics so far.
In China, Chinese medicine is used to prevent and treat people's flu and influenza is existing exceedes history in thousand, has accumulated rich experience.The medical Chinese medicine in treatment flu and influenza of people mainly is the antipyretic and antidotal type Chinese medicine and the apoplexy due to endogenous wind medicine that induces sweat; the former has Fructus Forsythiae, Flos Lonicerae, Radix Scutellariae, Herba Houttuyniae, Radix Isatidis, Folium Isatidis, Flos Chrysanthemi Indici, Folium Mori, Fructus Arctii, Herba Taraxaci, Calculus Bovis etc., and the latter has Rhizoma Zingiberis Recens, Ramulus Cinnamomi, Herba Ephedrae, Radix Bupleuri, Herba Schizonepetae, Folium Perillae, Flos Magnoliae, Herba Menthae etc.In addition, Chinese medicines such as the Radix Astragali, Radix Saposhnikoviae have remarkable promotion regulating action to human immunity, also are the control cold medicines of using always.The Chinese patent medicine that is used to prevent and treat people's flu is also various in style, more famous having " YINQIAO sheet ", " SHUANGHUANLIAN ", " QINGKAILING ", " Ramulus Cinnamomi Ephedrae Decoction " etc.
Along with developing rapidly of China's livestock and poultry breeding industry and deepening continuously of veterinary drug research, the application of Chinese medicine on livestock and poultry disease control and prevention is also more and more general, and survival rate and the health level that improves poultry has been played positive role.But as a whole, at present the herbal medicine of China is in also that compatibility of drugs lacks systematic study, product forms is coarse, effective ingredient is indeterminate, the low technical level of unstable product quality.Use the research of Chinese herb prevention poultry bird flu and report seldom, do not see the research report that has system deep, the herbal medicine product that bird flu is had fine prevention effect is not seen in veterinary drug market yet.
Summary of the invention
The object of the present invention is to provide a kind ofly has Chinese medicine composition of remarkable preventive effect and its production and application to poultry bird flus such as chicken, ducks.
To achieve these goals, the present invention takes following technical scheme:
The Chinese medicine composition of birds flu-preventing of the present invention, it is made up of effective ingredient and/or pharmaceutically acceptable carrier, and wherein said effective ingredient is made by following raw materials according: Fructus Forsythiae 90-110 weight portion, Flos Lonicerae 40-60 weight portion, northern Radix Isatidis 25-45 weight portion, Radix Scutellariae 40-60 weight portion, Rhizoma Dryopteris Crassirhizomatis 40-60 weight portion, Radix Astragali 70-90 weight portion.
Preferably, Fructus Forsythiae 100 weight portions, Flos Lonicerae 50 weight portions, northern Radix Isatidis 35 weight portions, Radix Scutellariae 50 weight portions, Rhizoma Dryopteris Crassirhizomatis 50 weight portions, the Radix Astragali 80 weight portions.More preferably Fructus Forsythiae 1000g, Flos Lonicerae 500g, northern Radix Isatidis 350g, Radix Scutellariae 500g, Rhizoma Dryopteris Crassirhizomatis 500g, Radix Astragali 800g.
Chinese medicine composition dosage form of the present invention is powder, drinking agent or dissolved granule.
The preparation method of Chinese medicine composition of the present invention comprises the steps:
(1) Fructus Forsythiae, Flos Lonicerae, northern Radix Isatidis, Radix Scutellariae, Rhizoma Dryopteris Crassirhizomatis, Radix Astragali pulverize separately are become coarse powder;
(2) get step (1) Fructus Forsythiae coarse powder and carry out the percolation extraction as solvent, collect percolate with ethanol water;
(3) get Flos Lonicerae coarse powder, northern Radix Isatidis coarse powder, Radix Scutellariae coarse powder, the Rhizoma Dryopteris Crassirhizomatis coarse powder that step (1) makes, make solvent, carry out percolation and extract, collect percolate with ethanol water;
(4) learn from else's experience radix astragali coarse powder that step (1) makes with the distillation water as solvent, carry out percolation and extract, collect percolate;
(5) merge the percolate of (2), (3), (4) step, leave standstill, filter, ethanol is reclaimed in distilling under reduced pressure, and residue medicinal liquid concentrating under reduced pressure under temperature 65-75 ℃, vacuum 0.02-0.04Mpa becomes thick paste, adds adjuvant and makes required dosage form.
Another preparation method of Chinese medicine composition of the present invention, comprise the steps: to take by weighing Fructus Forsythiae, Flos Lonicerae, northern Radix Isatidis, Radix Scutellariae, Rhizoma Dryopteris Crassirhizomatis, the Radix Astragali by the described percentage ratio consumption of claim 1, and pulverize separately becomes coarse powder, pressing the decocting cooking method decocts, collect the fried liquid of each time, after sedimentation is filtered, the weight that is evaporated to medicinal liquid under temperature 65-75 ℃, vacuum 0.02-0.04Mpa is 1.0 ± 0.1 times of Chinese medicine raw material gross weight, add 0.1% sodium benzoate and 0.05% nipalgin in the concentrated medicament and make antiseptic, promptly get drinking agent.
Chinese medicine composition of the present invention is used for the application of the medicine of birds flu-preventing in preparation.
The present invention at first utilizes " bird flu virus Embryo Gallus domesticus challenge test " (hereinafter to be referred as " Embryo Gallus domesticus test ") that kind of the heat-clearing and toxic substances removing surplus 40 and the apoplexy due to endogenous wind medicine that induces sweat are screened, and finds that kind of Chinese medicine has remarkable inhibitory action to bird flu virus in the Embryo Gallus domesticus surplus Rhizoma Dryopteris Crassirhizomatis, Fructus Forsythiae, Flos Lonicerae, Radix Scutellariae, Herba Houttuyniae, Radix Isatidis, Folium Isatidis, Radix Glycyrrhizae, Herba Hyperici perforati, the Radix Bupleuri etc. ten.Utilize the said medicine that filters out to carry out different prescriptions then, extract active ingredient with decocting in water and two kinds of methods of alcohol extraction respectively.At last compound recipe is carried out Embryo Gallus domesticus test and the therapeutic test of artificial onset's bird flu chicken, filter out effect compound recipe preferably.According to result of the test, determine that finally with 6 flavor Chinese medicines such as Fructus Forsythiae, Flos Lonicerae, northern Radix Isatidis, Radix Scutellariae, Rhizoma Dryopteris Crassirhizomatis, the Radix Astragali be that raw material carries out prescription, this side has remarkable prevention effect to chicken, duck bird flu.Fang Zhongqian 5 flavor medicines are the key medicine of heat-clearing and toxic substances removing, and bird flu virus is all had strong inhibitory action; Each medicine synergism of distinguishing the flavor of has not only been strengthened the inhibitory action to virus, antibacterial, and has been reduced the toxicity of single medicinal material and the generation of virus drug resistance.Wherein, the Fructus Forsythiae master contains phenethanol caffeic acid glycoside compound, and the Rhizoma Dryopteris Crassirhizomatis master contains pannol, the trident phenol of fainting, the Flos Lonicerae master contains chlorogenic acid, the Radix Isatidis master contains indigo, indirubin, and the Radix Scutellariae master contains baicalin, wogonin, and these compositions are to antibacterial and the viral inhibition activity that all has in various degree.The Radix Astragali contains astragalus polysaccharides, and astragalus polysaccharides has significant facilitation to the immunocompetence of humans and animals, can obviously strengthen the phagocytic function of macrophage, increases the TNF secretion, promotes the generation of antiviral antibody.
The specific embodiment
Embodiment 1
The preparation method of powder of the present invention comprises the steps:
(1) take by weighing Fructus Forsythiae 10000g, Flos Lonicerae 5000g, northern Radix Isatidis 3500g, Radix Scutellariae 5000g, Rhizoma Dryopteris Crassirhizomatis 5000g, Radix Astragali 8000g, and pulverize separately becomes coarse powder;
(2) get the Fructus Forsythiae coarse powder and make solvent with 80% ethanol, carry out percolation according to the fluid extract percolation, 2.5ml/kg.min collects percolate;
(3) Flos Lonicerae coarse powder, northern Radix Isatidis coarse powder, Radix Scutellariae coarse powder, the Rhizoma Dryopteris Crassirhizomatis coarse powder that step (1) makes of learning from else's experience made solvent with 60% ethanol, carries out percolation according to the fluid extract percolation, and 3ml/kg.min collects percolate;
(4) learn from else's experience radix astragali coarse powder that step (1) makes with the distillation water as solvent, carry out percolation according to the fluid extract percolation, 3ml/kg.min collects percolate;
(5) percolate in merging (2), (3), (4) step leaves standstill, and filters distilling under reduced pressure
Reclaim ethanol, residue medicinal liquid concentrating under reduced pressure under 65 ℃ of temperature, vacuum 0.02Mpa becomes thick paste, and spray drying or drying under reduced pressure were pulverized 60 mesh sieves, got dry fine powder;
(6) add 1.5% white carbon in the dry fine powder of step (5), promptly get powder.Wherein the effect of white carbon is to improve free-running property, reduces moisture absorption.Powder of the present invention is good powder of free-running property or powdery granule, is brown or sepia, bitter in the mouth.
Embodiment 2
The preparation method of dissolved granule of the present invention comprises the steps:
(1) take by weighing Fructus Forsythiae 9000g, Flos Lonicerae 5000g, northern Radix Isatidis 3500g, Radix Scutellariae 5000g, Rhizoma Dryopteris Crassirhizomatis 5000g, Radix Astragali 7000g, and pulverize separately becomes coarse powder;
(2) get the Fructus Forsythiae coarse powder and make solvent with 70% ethanol, carry out percolation according to the fluid extract percolation, 2.5ml/kg.min collects percolate;
(3) Flos Lonicerae coarse powder, northern Radix Isatidis coarse powder, Radix Scutellariae coarse powder, the Rhizoma Dryopteris Crassirhizomatis coarse powder that step (1) makes of learning from else's experience made solvent with 50% ethanol, carries out percolation according to the fluid extract percolation, and 2.5ml/kg.min collects percolate;
(4) learn from else's experience radix astragali coarse powder that step (1) makes with the distillation water as solvent, carry out percolation according to the fluid extract percolation, 3ml/kg.min collects percolate;
(5) merge the percolate of (2), (3), (4) step, leave standstill, filter, ethanol is reclaimed in distilling under reduced pressure, and it is 1.37 thick paste that residue medicinal liquid concentrating under reduced pressure under 75 ℃ of temperature, vacuum 0.03Mpa becomes relative density;
(6) an amount of to the thick paste adding cane sugar powder of step (5), sucrose accounts for 28% of granule finished weight, and mixing is made granule, and oven drying promptly gets dissolved granule.
Embodiment 3
The preparation method of drinking agent of the present invention comprises the steps:
(1) take by weighing Fructus Forsythiae 1100g, Flos Lonicerae 500g, northern Radix Isatidis 300g, Radix Scutellariae 600g, Rhizoma Dryopteris Crassirhizomatis 400g, Radix Astragali 800g, and pulverize separately becomes coarse powder;
(2) get the Fructus Forsythiae coarse powder and make solvent with 70% ethanol, carry out percolation according to the fluid extract percolation, 2.5ml/kg.min collects percolate;
(3) Flos Lonicerae coarse powder, northern Radix Isatidis coarse powder, Radix Scutellariae coarse powder, the Rhizoma Dryopteris Crassirhizomatis coarse powder that step (1) makes of learning from else's experience made solvent with 50% ethanol, carries out percolation according to the fluid extract percolation, and 2.5ml/kg.min collects percolate;
(4) learn from else's experience radix astragali coarse powder that step (1) makes with the distillation water as solvent, carry out percolation according to the fluid extract percolation, 3.0ml/kg.min collects percolate;
(5) merge percolate, leave standstill, filter, ethanol is reclaimed in distilling under reduced pressure, and residue medicinal liquid concentrating under reduced pressure under 65 ℃ of temperature, vacuum 0.02Mpa gets the concentrated medicament that weight is 3700g;
(6) add 0.1% sodium benzoate and 0.05% nipalgin in the concentrated medicament of step (5) and make antiseptic, promptly get drinking agent.
Embodiment 4
Another preparation method of drinking agent of the present invention comprises the steps:
Take by weighing Fructus Forsythiae 1050g, Flos Lonicerae 400g, northern Radix Isatidis 450g, Radix Scutellariae 400g, Rhizoma Dryopteris Crassirhizomatis 600g, Radix Astragali 700g, and pulverize separately becomes coarse powder; Press the decocting cooking method and decoct, collect the fried liquid of each time, after sedimentation was filtered, the weight that is evaporated to medicinal liquid under 72 ℃ of temperature, vacuum 0.04Mpa was 3600g, and the weight of this concentrated medicament all is feasible in the 3240g-3960g scope.Add 0.1% sodium benzoate and 0.05% nipalgin in the concentrated medicament and make antiseptic, promptly get drinking agent.
Embodiment 5: powder of the present invention
According to embodiment 1 described method, difference is to select following raw materials in weight for use: Fructus Forsythiae 9500g, Flos Lonicerae 4500g, northern Radix Isatidis 3000g, Radix Scutellariae 5500g, Rhizoma Dryopteris Crassirhizomatis 6000g, Radix Astragali 8500g.
Embodiment 6: dissolved granule of the present invention
According to embodiment 2 described methods, difference is to select following raw materials in weight for use: Fructus Forsythiae 10500g, Flos Lonicerae 5500g, northern Radix Isatidis 4000g, Radix Scutellariae 4500g, Rhizoma Dryopteris Crassirhizomatis 5000g, Radix Astragali 7500g.
Embodiment 7: drinking agent of the present invention
According to embodiment 3 described methods, difference is to select following raw materials in weight for use: Fructus Forsythiae 920g, Flos Lonicerae 420g, northern Radix Isatidis 260g, Radix Scutellariae 580g, Rhizoma Dryopteris Crassirhizomatis 520g, Radix Astragali 710g.
Embodiment 8: drinking agent of the present invention
According to embodiment 4 described methods, difference is to select following raw materials in weight for use: Fructus Forsythiae 1080g, Flos Lonicerae 580g, northern Radix Isatidis 430g, Radix Scutellariae 430g, Rhizoma Dryopteris Crassirhizomatis 590g, Radix Astragali 870g.
Medicine of the present invention can adopt the conventional method of Chinese medicine preparation to be prepared into any conventional innerwear Chinese medicine preparation.For example, ultra micro powder, tablet etc.But for the ease of giving chicken, the medication of duck colony, the preferred drinking agent of the present invention, dissolved granule, powder etc.Powder is made feedstuff and is added medication, and drinking agent and dissolved granule are made drinking-water and added medication.But these can not be used to limit protection scope of the present invention.
The consumption of Chinese medicine composition birds flu-preventing of the present invention is respectively:
(1) powder is used to prevent chicken, duck bird flu, and the addition in feedstuff is that feedstuff 1500 grams per ton are to 2000 grams; Be used for the treatment of chicken, duck bird flu, the addition in feedstuff is that feedstuff 2500 grams per ton are to 3000 grams;
(2) drinking agent is used to prevent chicken, duck bird flu, and addition is that the per 1000 gram body weight of chicken, duck are used drinking agent 3-5ml every day in an amount of drinking-water; Be used for the treatment of chicken, duck bird flu, addition is that the per 1000 gram body weight of chicken, duck are used drinking agent 6-8ml every day in an amount of drinking-water;
(3) dissolved granule is used to prevent chicken, duck bird flu, and addition is that the per 1000 gram body weight of chicken, duck restrain with electuary 0.4-0.6 every day in an amount of drinking-water; Be used for the treatment of chicken, duck bird flu, addition is that the per 1000 gram body weight of chicken, duck restrain with electuary 0.7-0.9 every day in an amount of drinking-water.
Below by test animal acute toxicity that example further specifies the Chinese medicine reconsideration preparation of birds flu-preventing of the present invention, drug effect and clinical before to the prevention effect of bird flu chicken.
Test one: the animal acute toxicity test of Chinese medicine composition
Materials and methods
Animal: Kunming mouse, body weight are 20.3 ± 1.2g, male and female half and half.Buy from Military Medical Science Inst., Guangzhou Military Region Rear-service Departmen.
Medicine: the Chinese medicine composition dissolved granule, Agricultural University Of South China's veterinary college Guangdong Province veterinary drug development is developed with safety evaluation key lab and is provided.
(1) mice LD
50Measure
60 of mices are divided into 6 groups at random, and 10 every group, feed the method administration with irritating, dosage and concentration are auspicious to see Table 1, and dose ratio is 1: 0.8 between group, and agent was apart from 1: 3, and mice is normal behind the single administration, observes continuously 7 days, and mice all survives, and can't record LD
50
(2) mtd test
Get 30 of mices, observe and test after three days, water is can't help in test fasting the previous day, under 25 ± 2 ℃ of conditions of room temperature, to irritate and feed once, drug level is 0.5g/ml (a maximum suspendible concentration), dosage is 1ml/20g (maximum volume), administration accumulative total is 25g/kg in one day, after the administration, and two weeks of breeding observing.
Viewing duration after the administration, mice diet, feces, fur are normal, and vegetative activity is all right, and none animal dead cuts open inspection and observes the main organs no abnormality seen.
Table 1 Chinese medicine composition drinking agent LD
50Measurement result
| Group | Number of animals | Drug level g/ml | Administration volume ml | Dosage g/kg | The dead mouse number | The survival mice situation |
| 1 | ?10 | ?0.16 | ?0.5 | ?4.00 | ?0 | Normally |
| 2 | ?10 | ?0.20 | ?0.5 | ?5.00 | ?0 | Normally |
| 3 | ?10 | ?0.26 | ?0.5 | ?6.50 | ?0 | Normally |
| 4 | ?10 | ?0.32 | ?0.5 | ?8.00 | ?0 | Normally |
| 5 | ?10 | ?0.40 | ?0.5 | ?10.00 | ?0 | Normally |
| 6 | ?10 | ?0.50 | ?0.5 | ?12.50 | ?0 | Normally |
Discuss and brief summary
Maximum suspendible concentration (0.50g/ml) with medicine is irritated hello administration (dosage is 12.5g/kg), can't measure LD
50So, measure its maximum tolerated dose.The maximum volume of (0.5g/ml) (1ml) carries out determination of acute toxicity under the Cmax that mice can be accepted, and mainly observes the untoward reaction in the two weeks.Mice is that viewing duration does not see that mice has untoward reaction under the maximum drug dose of 25g/kg in dosage as a result, and postmortem does not see that main organs has ANOMALOUS VARIATIONS yet, and the basic avirulence of this medicine of results suggest is clinical safe in utilization.
Test two: Chinese medicine composition oral liquid pharmacodynamics test
1. test material
1.1 test chicken: the yellow chicken of high-quality in 50 day age.Buy from poultry institute Guangdong, Guangdong Province fowl company during 1 day age, begin test raising to 50 day age.Adopt hemagglutination inhibition test (HI test) (HI) mensuration bird flu virus (AIV) negative 40 day age chicken.
1.2 feedstuff: buy from animal science institute of Agricultural University Of South China test chicken house, do not add any medicine.
1.3 medicine: used Chinese crude drug raw material is all bought from Guangzhou medical material company, and Agricultural University Of South China's veterinary college Guangdong Province veterinary drug development is extracted with safety evaluation key lab and preparation Chinese medicine composition drinking agent (oral liquid).Every milliliter of oral liquid is equivalent to the 1g crude drug.Amantadine is available from Hua Tian company of Agricultural University Of South China, and rimantadine is available from Zhejiang Yu Kang pharmaceutical Co. Ltd, and two kinds of medicines are all consistent with literature value with mass spectral analysis through fusing point test.Amantadine and rimantadine directly dissolve with distilled water, are made into mass concentration and are 0.2% medicinal liquid.
1.4 counteracting toxic substances strain: (half infects concentration 1.0 * 10 to H5 blood serum subtype bird flu virus
-6), Agricultural University Of South China veterinary college fowl diseases research department preserves.
2. test method
The test grouping: 90 of the yellow chickens of high-quality in 50 day age, male and female half and half are divided into 5 groups at random, 10 of normal control groups, all the other each groups are 20, and grouping situation in detail sees Table 2.
Counteracting toxic substances: the chick embryo allantoic liquid dilution that will contain bird flu virus is 0.5 * 10
-5Concentration, every chicken abdominal muscles injection 0.2ml inoculation.
Administration: each group of test administration was irritated and is fed administration on inoculation AIV same day, and each group of administration is all irritated and fed 5ml, and normal healthy controls group and infect matched group and irritate and feed normal saline is irritated and feed 1 time every day, and continuous irrigation was fed 7 days.
The evaluating drug effect standard: test duration 21 days, observe death condition and the clinical manifestation of chicken every day, the chicken that dies of illness carries out necropsy, and every chicken is weighed before and after the test.By following standard drug effect is estimated: (1) death: all at duration of test, bird flu classical symptom and dead appears, and it is dead that necropsy has promptly being judged to be of typical cytopathic feature to infect.(2) effective: after the off-test, all after administration still chickens of survival all be judged to be effectively, the percentage rate that every group of chicken number of surviving accounts for the test chicken number during off-test is an effective percentage.(3) weightening finish: the body weight of every chicken during according to on-test and during off-test, calculate the average weight and the standard deviation of the weightening finish of every chicken, every group of test chicken.The relative weight gain rate is to calculate by the ratio of each medication treatment group with the average weight gain of normal healthy controls group, and wherein the normal healthy controls group is made as 100%.
3. animal test results
3.1 infection matched group
Have in 20 infected chickens 17 dead in inoculation bird flu virus 15 days, 3 to the 21st day still healthy survivals of survival.Wherein behind the virus inoculation the 3rd day dead 2, the 4th day dead 3, the 4th day dead 5, the 5th day to the 10th day dead 5, the 11st day to the 15th day dead 2.Cardinal symptom: spirit is depressed, and appetite descends or is useless exhausted; Feather pine disorderly, the eye liquid of shedding tears; Draining sample feces; Be reluctant to walk about, severe patient does not rise with crouching, nervous symptoms such as curved neck is arranged, spin, jump.The necropsy feature: there is mucus in the oral cavity, and a plurality of organ enlargements such as liver, kidney, gallbladder also have the petechia, and the petechia is atropurpureus usually.
3.2 administration group
Infected chicken is behind drug treatment, and the Chinese medicine composition group has only 1 death, and survival rate is 95%, and clinical symptoms does not appear in most of chicken, indivedual chickens go out to lose face shed tears, feather pine slight disease such as disorderly, but in test symptom complete obiteration in the 21st day.Dead 18 of amantadine group chicken, dead 15 of rimantadine group chicken.Specifically see Table 2.
Three kinds of medicines of table 2 are to the prevention effect of avian influenza chicken
| Group | Quantity | Handle | Death toll | Mortality rate % | Effective percentage % | 50d body weight m/g | 71d body weight m/g | Rate of body weight gain % | The relative weight gain rate % |
| The normal healthy controls group | 10 | Do not infect, not administration | 0 | ?0 | ?- | ?685.3± ?58.7 | ?1103.6 ?±63.0 | ?61.04 | ?100 |
| Chinese medicine compound system group | 20 | Infection, 5.0ml oral liquid are irritated and are fed | 1 | ?5 | ?95 | ?683.9± ?63.7 | ?1091.2 ?±72.8 | ?59.56 | ?97.58 |
| The amantadine group | 20 | Infection, 10mg irritate and feed | 18 | ?90 | ?10 | ?692.4± ?67.5 | ?978.2± ?55.3 | ?41.28 | ?67.63 |
| The rimantadine group | 20 | Infection, 10mg irritate and feed | 15 | ?75 | ?25 | ?679.0± ?72.5 | ?982.0± ?81.6 | ?44.62 | ?73.10 |
| Infect matched group | 20 | Infection, not administration | 17 | ?85 | ?- | ?677.6± ?62.0 | ?971.6± ?35.4 | ?43.39 | ?71.10 |
4. conclusion
Result of the test shows; Chinese medicine composition of the present invention has good protection effect to the chicken of artificial challenge's bird flu virus; the survival rate that infects chicken reaches 95%; and the effective percentage of traditional anti-influenza virus medicament " amantadine ", " rimantadine " has only 10% and 25% respectively, and difference is extremely remarkable.The weightening finish of Chinese medicine composition group chicken is suitable with the normal healthy controls group, significantly is better than " amantadine " and " rimantadine " group, shows that Chinese medicine composition not only has good prevention effect to avian influenza, and the growth of chicken is had no adverse effects.
Test three: the Chinese medicine composition drinking agent is to the prevention effect test of cohabitation infection bird flu chicken
1. test material
1.1 test chicken: the yellow chicken of high-quality in 50 day age.Buy from poultry institute Guangdong, Guangdong Province fowl company during 1 day age, begin test raising to 50 day age.Adopt hemagglutination inhibition test (HI test) (HI) mensuration bird flu virus (AIV) negative 40 day age chicken.
1.2 feedstuff: buy from animal science institute of Agricultural University Of South China test chicken house, do not add any medicine.
1.3 medicine: used Chinese crude drug raw material is all bought from Guangzhou medical material company, and Agricultural University Of South China's veterinary college Guangdong Province veterinary drug development is extracted with safety evaluation key lab and preparation Chinese medicine composition drinking agent.Every milliliter of drinking agent is equivalent to the 1g crude drug.Rimantadine is available from Zhejiang Yu Kang pharmaceutical Co. Ltd, and fusing point test is consistent with literature value with mass spectrometry results.Rimantadine directly dissolves with distilled water, is made into mass concentration and is 1% medicinal liquid.
1.4 counteracting toxic substances strain: (half infects concentration 1.0 * 10 to H5 blood serum subtype bird flu virus
-6), Agricultural University Of South China veterinary college fowl diseases research department preserves.
2. test method
Artificial challenge's bird flu chicken: 50 of the yellow chickens of high-quality in 45 day age, male and female half and half, the chick embryo allantoic liquid dilution that will contain bird flu virus is 0.5 * 10
-5Concentration, every chicken abdominal muscles injection 0.2ml inoculation.The inoculation back began morbidity on the 3rd day, will have every day the morbidity chicken of obvious clinical symptoms to carry out labelling, and mean allocation is to each test chicken group, until inoculating back the 10th day.
The test grouping: the yellow chicken of age in days high-quality was 110 in 45 days, and male and female half and half are divided into 6 groups at random, 10 of normal control groups, and all the other each groups are 20, and grouping situation in detail sees Table 3.
Administration: began the administration of drinking water on the same day that has artificial challenge's bird flu chicken to change the test hurdle over to.Cut off the water supply 3 hours before the drinking-water administration.By the deal of every chicken 5ml, 10ml, 1% rimantadine is equipped with in the water fountain of 600ml tap water by the deal adding of every chicken 1ml, 2ml respectively the Chinese medicine composition drinking agent, every day 1 time, freely drinks water for chicken respectively.After the diseases caused by retention of fluid of adding medicine is intact, drink for chicken to tap water.Matched group is for the drink tap water.
The evaluating drug effect standard: test duration 28 days, observe death condition and the clinical manifestation of chicken every day, the chicken that dies of illness carries out necropsy, and every chicken is weighed before and after the test.The evaluating drug effect standard is with test two.
The malicious source of artificial challenge's confession chicken with live together the test chicken write down respectively, statistical disposition.
3. animal test results
3.1 cohabitation infection matched group
13 death in 28 days after changing artificial onset's bird flu chicken over to are arranged in 20 cohabitation infection chickens.Wherein the 3rd day dead 1, the 4th day dead 2, the 5th day dead 4, the 5th day to the 10th day dead 4, the 11st day to the 28th day dead 2.
3.2 administration group
The cohabitation infection chicken is behind drug treatment, and Chinese medicine composition 5ml dosage group is all survived, and 10ml dosage group has 1 death, and survival rate is 95%.Dead 9 and 11 respectively of rimantadine 10mg and 20mg dosage group chickens.Specifically see Table 3.The artificial challenge is all dead for malicious source chicken.
Table 3 Chinese medicine composition and rimantadine are to the prevention effect of cohabitation infection bird flu chicken
| Group | Quantity | Handle | Death toll | Mortality rate % | Survival rate % | 45d body weight m/g | 80d body weight m/g | Rate of body weight gain % | The relative weight gain rate % |
| The normal healthy controls group | 10 | Do not infect, not administration | 0 | ?0 | ?- | ?617.2 ?±45.3 | ?1356.3 ?±67.2 | ?119.8 | ?100 |
| Chinese medicine composition I group | 20 | Infection, a 5.0ml/ drinking agent | 0 | ?0 | ?100 | ?623.0 ?±60.5 | ?1327.0 ?±76.2 | ?113.0 | ?94.32 |
| Chinese medicine composition II group | 20 | Infection, a 10ml/ drinking agent | 1 | ?5 | ?95 | ?625.5 ?±56.6 | ?1330.5 ?±70.4 | ?112.7 | ?94.07 |
| Rimantadine I group | 20 | Infection, a 10mg/ drinking agent | 9 | ?45 | ?55 | ?612.7 ?±50.9 | ?1259.1 ?±63.2 | ?105.5 | ?88.06 |
| Rimantadine II group | 20 | Infection, a 20mg/ drinking agent | 11 | ?55 | ?45 | ?622.8 ?±68.7 | ?1179.0 ?±77.8 | ?89.31 | ?75.55 |
| Infect matched group | 20 | Infection, not administration | 13 | ?65 | ?35 | ?629.5 ?±47.2 | ?1297.6 ?±54.0 | ?106.1 | ?88.56 |
4. conclusion
The medical treatment result of the test of cohabitation infection shows; Chinese medicine composition of the present invention has the excellent protection effect to cohabitation infection bird flu chicken; supply the survival rate of 5ml and 10ml drinking agent to reach 100% and 95% respectively every every day, significantly is better than the protection effect of rimantadine.In addition, the chicken of cohabitation infection matched group is all morbidities not, and survival rate is 35%.
The weightening finish of Chinese medicine composition group chicken is poorer slightly than normal healthy controls group, but difference is not remarkable, and healthy relatively group is respectively 94.32% and 94.07%.Rimantadine has a significant effect to the growth of chicken, and 10mg and the weightening finish of 20mg dosage group descend 12% and 24% respectively than normal healthy controls group.The cohabitation infection test shows that Chinese medicine composition of the present invention has good prevention effect to natural infection bird flu chicken.
Claims (7)
1. the Chinese medicine composition of a birds flu-preventing, it is made up of effective ingredient and/or pharmaceutically acceptable carrier, it is characterized in that described effective ingredient made by following raw materials according: Fructus Forsythiae 90-110 weight portion, Flos Lonicerae 40-60 weight portion, northern Radix Isatidis 25-45 weight portion, Radix Scutellariae 40-60 weight portion, Rhizoma Dryopteris Crassirhizomatis 40-60 weight portion, Radix Astragali 70-90 weight portion.
2. Chinese medicine composition according to claim 1 is characterized in that described effective ingredient made by following raw materials according: Fructus Forsythiae 100 weight portions, Flos Lonicerae 50 weight portions, northern Radix Isatidis 35 weight portions, Radix Scutellariae 50 weight portions, Rhizoma Dryopteris Crassirhizomatis 50 weight portions, the Radix Astragali 80 weight portions.
3. Chinese medicine composition according to claim 1 is characterized in that described effective ingredient made by following raw materials according: Fructus Forsythiae 1000g, Flos Lonicerae 500g, northern Radix Isatidis 350g, Radix Scutellariae 500g, Rhizoma Dryopteris Crassirhizomatis 500g, Radix Astragali 800g.
4. Chinese medicine composition according to claim 1 is characterized in that its dosage form is powder, drinking agent or dissolved granule.
5. a method for preparing the described Chinese medicine composition of claim 1 is characterized in that comprising the steps:
(1) Fructus Forsythiae, Flos Lonicerae, northern Radix Isatidis, Radix Scutellariae, Rhizoma Dryopteris Crassirhizomatis, Radix Astragali pulverize separately are become coarse powder;
(2) get step (1) Fructus Forsythiae coarse powder and carry out the percolation extraction as solvent, collect percolate with ethanol water;
(3) get Flos Lonicerae coarse powder, northern Radix Isatidis coarse powder, Radix Scutellariae coarse powder, the Rhizoma Dryopteris Crassirhizomatis coarse powder that step (1) makes, make solvent, carry out percolation and extract, collect percolate with ethanol water;
(4) learn from else's experience radix astragali coarse powder that step (1) makes with the distillation water as solvent, carry out percolation and extract, collect percolate;
(5) merge the percolate of (2), (3), (4) step, leave standstill, filter, ethanol is reclaimed in distilling under reduced pressure, and residue medicinal liquid concentrating under reduced pressure under temperature 65-75 ℃, vacuum 0.02-0.04Mpa becomes thick paste, adds adjuvant and makes required dosage form.
6. method for preparing the described Chinese medicine composition of claim 1, it is characterized in that comprising the steps: taking by weighing Fructus Forsythiae by the described percentage ratio consumption of claim 1, Flos Lonicerae, the north Radix Isatidis, Radix Scutellariae, Rhizoma Dryopteris Crassirhizomatis, the Radix Astragali, and pulverize separately becomes coarse powder, pressing the decocting cooking method decocts, collect the fried liquid of each time, after sedimentation is filtered, at temperature 65-75 ℃, the weight that is evaporated to medicinal liquid under the vacuum 0.02-0.04Mpa is 1.0 ± 0.1 times of Chinese medicine raw material gross weight, add 0.1% sodium benzoate and 0.05% nipalgin in the concentrated medicament and make antiseptic, promptly get drinking agent.
7. Chinese medicine composition as claimed in claim 1 is used for the application of the medicine of birds flu-preventing in preparation.
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