CN104784657A

CN104784657A - Application of traditional Chinese veterinary medicine compound preparation for preparing medicine for treating chicken colibacillosis

Info

Publication number: CN104784657A
Application number: CN201510124327.4A
Authority: CN
Inventors: 赖小平; 倪晨; 黄松; 袁捷; 谢友良; 汪芳; 张军
Original assignee: Guangzhou University of Chinese Medicine
Current assignee: Guangzhou University of Chinese Medicine
Priority date: 2015-03-20
Filing date: 2015-03-20
Publication date: 2015-07-22
Anticipated expiration: 2035-03-20
Also published as: CN104784657B

Abstract

The invention discloses an application of a traditional Chinese veterinary medicine compound preparation for preparing a medicine for treating chicken colibacillosis, and particularly relates to an application of a traditional Chinese veterinary medicine compound preparation for preparing a medicine for treating chicken colibacillosis in subtropical climate regions such as Southern China. The traditional Chinese veterinary medicine compound preparation comprises the following components by percent: 5-55% of common andrographis herb, 10-65% of rhizoma atractylodis, 5-55% of radix aucklandiae, 5-55% of cortex magnoliae officinalis and 10-65% of rhizoma zingiberis. The traditional Chinese veterinary medicine compound preparation is powder. A preparation technology comprises the steps that the five traditional Chinese medicinal materials are crushed into coarse particles respectively, are mixed according to a prescription ratio, are further pulverized, pass through a 30-mesh sieve, are placed in a V-shaped mixer for mixing for 30min, and then are subpackaged. The traditional Chinese veterinary medicine compound preparation particularly has a very good curative effect on the chicken colibacillosis in high-temperature, rainy and hot-humid regions, can effectively control spreading and diffusion of the chicken colibacillosis, is safe and small in side effect, is significant for poultry husbandry in the subtropical climate regions such as Southern China, and has better economic and social significance.

Description

The application of a kind of herbal medicine compound preparation in the medicine of preparation treatment chicken colibacillosis

Technical field

The invention belongs to herbal medicine technical field.More specifically, the application of a kind of herbal medicine compound preparation in the medicine of preparation treatment chicken colibacillosis is related to.

Background technology

In recent years, along with the development of aviculture, chicken colibacillosis extensively betides poultry-farm in all parts of the country, and becomes a primary study object in veterinary clinic diseases prevention and treatment already.Chicken colibacillosis is common clinical, frequently-occurring disease, by number of ways infected chicken, cause chicken multiple organ inflammation and can cause death, this sick rapid onset, infectiousness are strong, carrying out huge economic loss to cultivation industrial belt, is one of infectious disease of harm aquaculture.

Current most of raiser prevents and treats colibacillary thinking and is still medicine and controls, and this disease of clinical treatment is main based on antimicrobial drug, but has R-plasmid due to escherichia coli, easily produce drug resistance, reduction curative effect of medication, therefore clinical therapeutic efficacy is poor; Simultaneously due to a large amount of, the Use out of range of antimicrobial drug, cause drug residues a large amount of in Carnis Gallus domesticus and egg, carry out great hidden danger to food seat belt.In addition, due to the impact of the comprehensive extraneous factors such as weather, for different regions poultry-farm, obvious diversity is also also existed to the control of chicken colibacillosis.

South China, especially South China, based on subtropical monsoon climate, annual precipitation is 1400 ~ 2000 millimeters, high temperature and rainy, damp and hot, aquaculture is chicken farm especially, and chicken colibacillosis all can occur throughout the year, every year rainy, sultry, wet season is multiple, prevention effect is poor, easily recurs.

In this context, especially for the aviculture in the subtropical climate areas such as China south China, develop the herbal medicine new drug of safe and effective, that side effect is little treatment chicken colibacillosis, there is good economic implications and social meaning.

Summary of the invention

The technical problem to be solved in the present invention is the defect and the deficiency that overcome existing chicken colibacillosis medicine, a kind of application of herbal medicine compound preparation in preparation control chicken colibacillosis medicine for the treatment of chicken colibacillosis is provided, especially for the application in the subtropical climate areas such as China south China, spreading and spreading of this disease is controlled.

Above-mentioned purpose of the present invention is achieved through the following technical solutions:

Treat the application of herbal medicine compound preparation in the medicine of preparation treatment chicken colibacillosis for chicken colibacillosis, the application especially in the medicine of preparation treatment subtropical climate area chicken colibacillosis.

Preferably, described subtropical climate area refers to South China of China.

Above-mentioned herbal medicine compound preparation is made up of the component of following percentage composition: 5 ~ 55% Herba Andrographis, 10 ~ 65% Rhizoma Atractylodis, 5 ~ 55% Radix Aucklandiae, 5 ~ 55% Cortex Magnoliae Officinalis, 10 ~ 65 % Rhizoma Zingiberiss.

The dosage form of the herbal medicine compound preparation of described treatment chicken colibacillosis is powder, and preparation technology is as follows: after five kinds of Chinese crude drugs are broken into coarse granule respectively, mix in prescription ratio, further fine powder is broken, crosses 30 mesh sieves, puts in V-Mixer and mixes 30 minutes, subpackage.

Preferably, the herbal medicine compound preparation of above-mentioned treatment chicken colibacillosis is made up of the component of following percentage composition: 10 ~ 40% Herba Andrographis, 15 ~ 55% Rhizoma Atractylodis, 10 ~ 40% Radix Aucklandiae, 10 ~ 40% Cortex Magnoliae Officinalis, 15 ~ 55% Rhizoma Zingiberiss.

More preferably, the herbal medicine compound preparation of above-mentioned treatment chicken colibacillosis is made up of the component of following percentage composition: 10 ~ 25% Herba Andrographis, 20 ~ 35% Rhizoma Atractylodis, 10 ~ 25% Radix Aucklandiae, 1 ~ 25% Cortex Magnoliae Officinalis, 20 ~ 35% Rhizoma Zingiberiss.

Most preferably, the herbal medicine compound preparation of above-mentioned treatment chicken colibacillosis is made up of the component of following percentage composition: Herba Andrographis 16.7%, Rhizoma Atractylodis 25%, the Radix Aucklandiae 16.7%, Cortex Magnoliae Officinalis 16.6%, Rhizoma Zingiberis 25%.

The herbal medicine compound preparation of above-mentioned treatment chicken colibacillosis is preparing the application in the medicine for the treatment of chicken diarrhea also within protection scope of the present invention, and described chicken diarrhea is the diarrhoea caused by Oleum Ricini or Folium Sennae.

The herbal medicine compound preparation of above-mentioned treatment chicken colibacillosis is preparing the application in suppression chicken or mice Intestinal pushing function medicament also within protection scope of the present invention.

The pharmaceutical characteristic of each active component of the present invention is as follows:

Herba Andrographis: heat-clearing and toxic substances removing, removing heat from blood are subsided a swelling.Cure mainly the cardiovascular disease such as acute bacillary dysentery, gastroenteritis, upper respiratory tract infection, cold, fever and hypertension.Modern medicine study shows, Herba Andrographis has obvious inhibitory action to avian pasteurella multocida, S. pullonum, escherichia coli, staphylococcus aureus, bacillus pyocyaneus etc., also has antiinflammatory, antivirus action simultaneously.On modern clinic, also conventional CHUANXINLIAN ZHUSHEYE treats diarrhoeal diseases.

Rhizoma Atractylodis: expelling wind and cold, the spleen strengthening and damp drying.Cure mainly distension and fullness in the abdomen, the retention of dampness in middle-JIAO card such as to have loose bowels.Modern pharmacological research shows, Rhizoma Atractylodis all have obvious inhibitory action to staphylococcus aureus, tulase, dead leaf bacillus and bacillus pyocyaneus, and the mice that causes of the mouse small intestine diarrhoea that Oleum Ricini is caused and Folium Sennae too enteral diarrhea have obvious anti-diarrhea effect.In addition, Rhizoma Atractylodis also have inhibitory action in various degree to the multiple fungus such as trichophyton, trichophyton gypseum.

The Radix Aucklandiae: promoting the circulation of QI to relieve pain, strengthening the spleen to promote digestion.The treatment of the digestive tract disease such as treating gastric ulcer, biliary colic, diarrhoea is the common medicine for the treatment of gastrointestinal disease.Modern pharmacological research shows, the Radix Aucklandiae can reduce mouse small intestine diarrhoea and large enteral diarrhea number of times, also has weak inhibitory action to mice prepared Chinese ink gastrointestinal propulsion.

Cortex Magnoliae Officinalis: dampness expectorant, the therapeutic method to keep the adverse QI flowing downwards are except full.Cure mainly humidity hysteresis damaging the spleen and stomach, the vomiting and diarrhoea of gastral cavity painful abdominal mass, stagnation of QI due to dyspepsia, abdominal distention constipation, phlegm retention breath with cough, have antiulcer and regulate the effect of digestive system.

Rhizoma Zingiberis: warming spleen and stomach for dispelling cold, recuperating depleted YANG is promoted blood circulation, dampness expectorant.Cure mainly enteritis, diarrhoea, vomiting etc.Modern pharmacological research shows, Rhizoma Zingiberis has the antibacterial Zuo ﹑ of the scorching Zuo ﹑ of analgesia Kang and improves the effects such as local blood circulation.

But, for the treatment of chicken colibacillosis in real life, not not merely apply the curative effect that can suppress colibacillary medicine just can to obtain, the integrated survey of the birds life habits such as chicken and physiological situation is also absolutely necessary, the effect that environmental factors is treated for chicken colibacillosis also has great impact, therefore composite for medicine, must consider the impact of environmental factors, just can reach optimum therapeutic effect.

The loose treatment being mainly used in chicken colibacillosis of lotus perfume of the present invention, especially for high temperature, rainy, damp-heat area, both selected Herba Andrographis heat-clearing and toxic substances removing, the dampness of bitter cold, compatibility Rhizoma Atractylodis are drying damp and strengthening spleen again; The Radix Aucklandiae, Cortex Magnoliae Officinalis circulation of qi promoting removing dampness, help with Rhizoma Zingiberis needle warming moxibustion with dehumidifying, and the property of his medicine bitter cold of assistant system is to protect gastric qi, and all medicines share, and play the merit of heat-clearing and toxic substances removing, spleen invigorating dehumidifying, antidiarrheal altogether.Because except turbid damp and antidiarrheal, except part of the body cavity below the umbilicus, housing the bladder, kidneys and bowels turbid damp improves tunica mucosa tubae uterinae hyperemia, rotten to the corn thinning, play antiinflammatory bacteriostasis efficacy.

The prescription composition of this aspect is scientific and reasonable: Herba Andrographis heat-clearing and toxic substances removing, dampness removing and diarrhea-relieving are monarch drug, and Rhizoma Atractylodis the spleen strengthening and damp drying is ministerial drug, and the Radix Aucklandiae, Cortex Magnoliae Officinalis circulation of qi promoting removing dampness are adjuvant drug, and dehumidifying that Rhizoma Zingiberis is needle warming moxibustion is for making medicine.Common except damp and hot, adjust taste, antidiarrheal dysentery.In side, Herba Andrographis is bitter, cold.Return large intestine channel, heat clearing away large intestine detoxifies, removing heat from blood dampness-eliminating and dysentery-stopping.

Rhizoma Atractylodis: pungent, bitter, temperature.Return taste warp. yE Tian shi" linzheng Zhinan Yi" say: " and husband's epidemic disease is foul gas, ancients thus drink fragrance, adopt Herba Chlorophyti, to attack the gas of fragrance also, heavily wash also dirty." " Mingyi Bielu " record Rhizoma Atractylodis " swollen by edema due to wind pathogen knot between skin, except spasm in the epigastrium is full and cholera is told incessantly lower, warming the stomach rapid digestion of food is addicted to food for main headache, expectorant water." Compendium of Material Medica: " control damp-phlegm prolonged fluid-retention ... spleen wets dirty, turbid drop leukorrhagia, lingering diarrhea hemorrhoidal hamorrhage." the bitter temperature of Rhizoma Atractylodis, the power of dry taste turbid damp wins especially, the pungent perfume (or spice) of its gas, can invigorating the spleen and regulating the stomach, to help disease chicken food for taking a turn for the better, reduces diarrhoea number of times.

The Radix Aucklandiae, pungent, bitter, temperature.Return taste, large intestine channel." medicineization justice " cloud: " Radix Aucklandiae, perfume can be ventilated, and closes the five internal organs, for adjusting all gas key medicines.Control the vexed belch of painful abdominal mass with this, edema abdominal distention, dysentery beriberi, all adjust the merit of stagnant loose gas.Therefore this product benefaction gastrointestinal stagnation of QI, intestinal stasis relieving pain relieving in tune, for the disease of weight after dysentery." " Japan hanako materia medica ": " and control all gas of trusted subordinate, antidiarrheal, cholera, dysentery, antiabortive, strengthening the spleen to promote digestion.Treat win bad, bladder cold type of pain, vomiting and regurgitation." " herbal classic ": " main pathogen, wards off malicious epidemic disease, Qiang Zhi, main pouring dew.”

Cortex Magnoliae Officinalis, bitter, pungent, temperature.Return spleen, stomach, large intestine channel." Mingyi Bielu ": " main warming middle-JIAO, QI invigorating, the expectorant therapeutic method to keep the adverse QI flowing downwards.Control cholera and stomachache, distension, cold in the stomach is inverse, and vomiting is more than in the heart, lets out dysentery, drenches dew.Except frightened, going or staying heat, only stuffy sensation with restlessness, thick the intestines and stomach." " book on Chinese herbal medicine just " cloud Cortex Magnoliae Officinalis can " temperature drop, falls apart stagnant, eliminating cold-damp dysentery." the bitter dry pungent row of this product, can dampness, again can circulation of qi promoting, benefaction gastrointestinal stagnation of QI distension." good recipe tested by collection " stops immediately watery diarrhea side, and " Shengji Zonglu " Cortex Magnoliae Officinalis soup etc. all wet to be contained to control with card compatibility with this product, cold-damp dysentery.Cortex Magnoliae Officinalis and the Radix Aucklandiae compatible, strengthen the merit of promoting the circulation of QI to relieve pain, improve the symptoms such as disease chicken poor appetite, diarrhoea.

Rhizoma Zingiberis, pungent, heat.Return spleen, stomach warp.This product is hot dry strong, and master enters taste and is longer than warming spleen and stomach for dispelling cold, strong activating the spleen sun, is the principal agent of warm middle Jiao.Shennong's Herbal: " hemostasis, perspires for main fullness in the chest cough with dyspnea, warming middle-JIAO, by rheumatism numbness, and intestinal Bleach dysentery.Survivor is especially good." " pearl sac ": " and Rhizoma Zingiberis its with having four: logical heart-yang, one also; Remove internal organs chronic diseases, two also; The cold sending out warp all, three also; Control the cold stomachache of sense, four also." side in add Rhizoma Zingiberis, both helped and controlled the cold of heat clearing away medicine, prevent and treat too cold and cool damage spleen sun, get again the effect of Rhizoma Zingiberis temperature fortune, make spleen wet easily remove.

Make a general survey of we, Herba Andrographis heat-clearing and toxic substances removing, Rhizoma Atractylodis dampness, the compatibility Radix Aucklandiae, Cortex Magnoliae Officinalis adjust smooth taste mechanism of qi, assistant, with needle warming moxibustion Rhizoma Zingiberis, had both helped the transporting of spleen, and had prevented heat-clearing and toxic substances removing again and do not hinder the foundation of acquired constitution, all medicines share plays heat-clearing and toxic substances removing, dampness circulation of qi promoting altogether, effect of solid protection taste, makes damp and hotly easily to remove, and gastrointestinal mechanism of qi is adjusted suitable, strengthen disease chicken immunity, alleviate escherichia coli disease symptoms.

In sum, the selection of this prescription has sufficient Chinese veterinarian's theoretical foundation: chicken colibacillosis is mainly in hot climate wet season, damp and hot inclined Sheng, chicken experiences Shi Re Bing Pian morbidity, pathogen is by the change of disease of epidemic febrile disease defensive-qi-nutrient-blood, and syndrome of WEI system performance feather is fluffy and disorderly, lethargy; Damp invasion of lower energizer large intestine, therefore performance aquiform stool and loose stool.Damp and hotly import edema caused by disorder of QI into, performance heating, breathe and breathe heavily thick, injure the five internal organs, be easy to dead.Severe patient pathogenic heat the pathogen attacking the pericardium adversely or go deep into blood system, we lotus perfume is loose tightly keeps above etiology and pathogenesis, when not falling ill, timely heat clearing and damp drying, dispel damp-heat disease because preventing morbidity, compatibility spleen invigorating circulation of qi promoting method for the treatment of, the gastrointestinal function of adjustment and enhancing chicken, improve immunity and the resistance of chicken self, thus reduce the sickness rate of colibacillosis.Strive prevention of diseae.In the morbidity course of disease, defend stage by stage, damp and hot Yong Sheng, incoordination between the spleen and stomach, large intestine conduction is not normal, shows as listless, and feed intake declines, feather is fluffy and disorderly, heat-clearing and toxic substances removing, dampness early when aquiform stool and loose stool, wetly do not go then heat not remove, regulating gastointestinal mechanism of qi simultaneously, prevent and treat damp and hot pathogen and enter the inner change of disease further, reduce the infringement after Colibacillosis.In addition, we inheriting damp eliminating heat clearing away, defend on basis that gas rules together, strengthen the power of dampness, and combine the protecting stomach-QI principle of " taste are ruled together ", strengthen and help strong fortune chicken spleen and stomach function Chinese medicine, with strengthening vital QI to eliminate pathogenic factors, treating both the principal and the secondary aspects of a disease at the same time.

The present invention has following beneficial effect:

The invention discloses the application of a kind of herbal medicine compound preparation in the medicine of preparation treatment chicken colibacillosis, this prescription Chinese medicine has extraordinary curative effect to chicken colibacillosis, effectively can control spreading and spreading of this disease, and safety, side effect are little, application prospect is good.

Experimental result shows, theoretical for foundation with sufficient Chinese veterinarian, and by reasonably composite, in South China of China, the curative effect of prescription Chinese medicine of the present invention to chicken colibacillosis is better than contrast medicine four taste Herba Andrographis and falls apart.Especially to high temperature, rainy, damp-heat area, therapeutic effect is good, speed is fast, and safety, side effect are little, can reach the effect for the treatment of both the principal and secondary aspects of a disease, significant to the aviculture in the subtropical climate areas such as China south China, have good economic implications and social meaning.

Accompanying drawing explanation

Fig. 1 is preparation technology's flow chart that lotus perfume is loose.

Detailed description of the invention

Further illustrate the present invention below in conjunction with Figure of description and specific embodiment, but embodiment does not limit in any form to the present invention.Unless stated otherwise, the present invention adopts reagent, method and apparatus are the art conventional reagent, method and apparatus.

Unless stated otherwise, agents useful for same of the present invention and material are commercial.

embodiment 1 the present invention treats preparation method and the lab scale craft of the herbal medicine compound preparation of chicken colibacillosis

1, preparation method

(1) prescription name: lotus perfume is loose.

Prescription: Herba Andrographis 167g, Rhizoma Atractylodis 250g, Radix Aucklandiae 167g, Cortex Magnoliae Officinalis 166g, Rhizoma Zingiberis 250g, makes 1000g.

(2) preparation method: above five kinds of Chinese crude drugs, are broken into coarse granule respectively, then in the mixing of prescription ratio, pulverize further, cross 30 mesh sieves, put in V-Mixer and mix 30 minutes, subpackage.

Dosage form: the dosage form of this herbal medicine compound preparation is powder.

(3) this lotus perfume is loose is made up of Herba Andrographis, Rhizoma Atractylodis, the Radix Aucklandiae, Cortex Magnoliae Officinalis and Rhizoma Zingiberis 5 taste medical material, wherein Rhizoma Atractylodis, the Radix Aucklandiae, Rhizoma Zingiberis 3 taste medical material are mainly containing volatile oil, inventor has investigated this 3 taste medical material water extraction and has received oily technique, found that oil yield lower (being about 0.53%); And the andrographolide in principal agent Herba Andrographis medical material is by thermally labile, is easily converted into dehydrorographolide, therefore determine that directly medical material being beaten powder is used as medicine.

2, lab scale craft

(1) flour extraction is investigated

Take the clean medical material of Herba Andrographis, Rhizoma Atractylodis, the Radix Aucklandiae, Cortex Magnoliae Officinalis and Rhizoma Zingiberis by recipe quantity, weigh 3 parts, every part of 10kg, pulverize, cross 30 mesh sieves, calculate flour extraction.Result is as shown in table 1.

The loose flour extraction of table 1 lotus perfume

(2) research before preparation prescription: the fragrant loose powder of lotus broken mistake 30 order, measure mobility, hygroscopicity, bulk density, granularity etc., result is as shown in table 2.Result shows, and the loose mobility of 30 order lotus perfume is better, and hygroscopicity is not strong.

The character of the loose raw material of table 2 lotus perfume

(3) adjuvant is selected: according to the character that lotus perfume is loose, its mobility is better, and hygroscopicity is not strong, add and use screw rod powder subpackage, less demanding to mobility, therefore need not use fluidizer, medicated powder gas is fragrant, bitter in the mouth, after mixed feeding, bitterness is not clearly, and chicken feels not obvious to bitterness, do not affect the feed of chicken, therefore need not add the adjuvant such as correctives, fluidizer.

(4) process route is selected: according to Chinese powder medicine preparations shaping process route, determines the loose production process route of lotus perfume as shown in Figure 1.

(5) preparations shaping technical study: lotus perfume (or spice) is dispersed in raw medicinal material pre-treatment research have been pulverized and sieved, and therefore preparations shaping technique mainly mixes and divided dose.

It is V-Mixer that powder mixes the most frequently used mixing apparatus, therefore uses V-Mixer to test, below tests the relation of paper examines incorporation time and mixing homogeneity and uniformity of dosage units, thus determine incorporation time.

The investigation of incorporation time: take the fragrant divided powder 20kg of lotus, mix 10 minutes at laboratory V-Mixer respectively, 20 minutes, 30 minutes time carry out sampling (sample point is respectively in the sampling of upper, middle and lower each point), check appearance uniform degree.Result is as shown in table 3.

Table 3 incorporation time investigates result

As shown in Table 3, mix 10 minutes, 20 minutes, 30 minutes appearance uniform degree qualified, appearance uniform degree does not have too big-difference, uniformity of dosage units and production feed intake scale and equipment relevant, follow-up Pilot scale runs shows, mixing 30 minutes uniformity of dosage units can be qualified, therefore considers the suitability of technique, and incorporation time selects be advisable for 30 minutes.

(6) subpackage: the lotus perfume getting mix homogeneously is loose, equipment debugging accurately after subpackage 30 bags, check by Minimum loading capacity examination method, all meet requirement under powder item.

3, process certification test

In process certification test, by the loose fixed recipe quantity of lotus perfume and technique, long run test 3 batches.Namely the fragrant loose powder of lotus is broken, is investigate project, the results are shown in Table 4 with flour extraction.

Table 4 lotus fragrant day labor skill demonstration test result

Result shows: adopt the loose flour extraction average of this technique lotus perfume to be 97.6%, technique favorable reproducibility, process stabilizing.

In sum, according to above research, determine that the preparation method that lotus perfume is loose is: get Herba Andrographis, Rhizoma Atractylodis, the Radix Aucklandiae, Cortex Magnoliae Officinalis, Rhizoma Zingiberis five kinds of clean medical materials, be ground into coarse granule respectively, then in the mixing of prescription ratio, further fine powder is broken, cross 30 mesh sieves, put in V-Mixer and mix 30 minutes, subpackage, get final product (concrete process for producing as shown in Figure 1).

the loose quality standard of embodiment 2 lotus perfume

[ prescription name] the loose Lian xiang san of lotus perfume

[ prescription] Herba Andrographis 167g, Rhizoma Atractylodis 250g Radix Aucklandiae 167g Cortex Magnoliae Officinalis 166g Rhizoma Zingiberis 250g.

[ method for making] above gomi herbs, pulverize, sieve, mixing, to obtain final product.

[ character] this product is lark powder; Gas is fragrant, bitter in the mouth.

[ function] heat-clearing and toxic substances removing, except humidifying stagnant.

[ cure mainly] for chicken colibacillosis.

[ specification] 250g/ bag.

[ usage and consumption] mixed feeding: every 1kg feedstuff, chicken 10g.

[ storage] sealing, moistureproof.

[ effect duration] 2 years.

[ differentiate] (1) get this product 6g, add methanol 100mL, soaked overnight, supersound process (power 360 W, frequency 40 kHz) 30min, filter, filtrate 60 DEG C of evaporates to dryness, the residue 15mL that adds water makes dissolving, cross AB-8 type macroporous adsorbent resin (post footpath 1.3cm, post height 15cm), wash with water to colourless, use 50mL 40% ethanol more respectively, 100mL 70% ethanol gradient elution, collect 70% ethanol elution, be evaporated to without alcohol taste, add water 20mL, 3 times are extracted with chloroform jolting, each 20mL, merge chloroform liquid, 60 DEG C of evaporates to dryness, residue adds ethyl acetate 1mL makes dissolving, as need testing solution.Separately get Herba Andrographis control medicinal material 2 g, add ethanol 30 mL, supersound process (power 360 W, frequency 40 kHz) 30 minutes, filter, filtrate 60 DEG C of evaporates to dryness, residue adds ethanol 2 mL makes dissolving, medical material solution in contrast.Test according to thin layer chromatography (" Chinese veterinary pharmacopoeia " 2010 editions two annex 32 pages), draw above-mentioned need testing solution 10 μ L, control medicinal material solution 8 μ l, put respectively on same GF254 lamellae, with chloroform-acetate-methanol (4:3:0.4) for developing solvent, launch, take out, dry, inspect under putting ultra-violet lamp (254 nm).In test sample chromatograph, on the position corresponding to control medicinal material chromatograph, the fluorescence speckle of aobvious same color.

(2) get this product powder 6g, add methanol 50mL, supersound process (power 360 W, frequency 40 kHz) 30 minutes, filter, filtrate reduced in volume is to thick paste, and the 30mL that adds water dissolves, 3 times are extracted with chloroform jolting, each 20 mL, merge chloroform liquid, 50 DEG C of evaporates to dryness, residue adds methanol 5mL makes dissolving, as need testing solution.Separately get Rhizoma Atractylodis control medicinal material 1g, add methanol 10mL, supersound process (power 360 W, frequency 40 kHz) 15 minutes, filter, get filtrate, in contrast medical material solution.Test according to thin layer chromatography (" Chinese veterinary pharmacopoeia " 2010 editions two annex 32 pages), draw need testing solution 6 μ L, control medicinal material solution 8 μ L, put respectively on same silica gel g thin-layer plate, with cyclohexane-ethyl acetate (22:1) for developing solvent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, and 105 DEG C to be heated to spot development clear.In test sample chromatograph, on the position corresponding to control medicinal material chromatograph, the speckle of aobvious same color.

(3) [discriminating] 2 lower need testing solutions are got as need testing solution.Separately get Radix Aucklandiae control medicinal material 1g, add methanol 20mL, supersound process (power 360 W, frequency 40 kHz) 30 minutes, filter, 50 DEG C of evaporates to dryness, residue adds methanol constant volume to 2mL, in contrast medical material solution.Separately get dehydrocostuslactone reference substance, costunolide reference substance, add methanol and make the solution of every 1mL containing 0.5mg respectively, product solution in contrast.Test according to thin layer chromatography (" Chinese veterinary pharmacopoeia " 2010 editions two annex 32 pages), draw need testing solution 3 μ L, control medicinal material solution 4 μ L under [discriminating] 2, two kinds of each 6 μ L of reference substance solution, put respectively on same silica gel g thin-layer plate, with toluene-methanol (27:1) for developing solvent, launch, take out, dry, spray is with 5% vanillin-sulfuric acid solution, and 105 DEG C to be heated to spot development clear.In test sample chromatograph, on the position corresponding with reference substance chromatograph to control medicinal material, the speckle of aobvious same color.

(4) get this product powder 6g, add methanol 50mL, supersound process (power 360 W, frequency 40 kHz) 30 minutes, filter, filtrate evaporate to dryness, residue adds dilute hydrochloric acid 40mL and dissolves, and extracts 3 times with chloroform jolting, each 20mL, merges chloroform liquid, extracts 3 times with 2%NaOH jolting, each 20mL, merges alkaline solution, adds hydrochloric acid and adjusts pH to 1 ~ 2, extract 3 times with chloroform jolting, each 20mL, merge chloroform liquid, evaporate to dryness, residue adds methanol 1mL and dissolves, as need testing solution.Separately get Cortex Magnoliae Officinalis control medicinal material 1g, add methanol 20mL, close plug, supersound process (power 360 W, frequency 40 kHz) 30 minutes, filter, evaporate to dryness, residue adds methanol constant volume to 2mL, in contrast medical material solution.Test according to thin layer chromatography (" Chinese veterinary pharmacopoeia " 2010 editions two annex 32 pages), draw above-mentioned need testing solution 5 μ L, control medicinal material solution 3 μ L, put respectively on same silica GF254 lamellae, with cyclohexane-acetone (7:2) for developing solvent, launch, take out, dry, inspect under putting ultra-violet lamp (254 nm).In test sample chromatograph, on the position corresponding to control medicinal material chromatograph, the fluorescence speckle of aobvious same color.

[ check] powder item should be met under relevant every regulation (" Chinese veterinary pharmacopoeia " 2,010 two annex 5 pages).

[ assay] andrographolide, dehydrorographolide, costunolide and dehydrocostuslactone content according to high performance liquid chromatography " Chinese veterinary pharmacopoeia " 2010 editions two annex 34 pages of mensuration.

Chromatographic condition and system suitability: be filler with octadecylsilane chemically bonded silica, take methanol as mobile phase A, water is Mobile phase B, carries out gradient elution according to table 5; Dehydrorographolide determined wavelength is 254 nm, and andrographolide, costunolide, dehydrocostuslactone all detect under 225 nm.Theoretical cam curve is not less than 3000.

Table 5 gradient elution table

The preparation of reference substance solution: get andrographolide, dehydrorographolide, costunolide and dehydrocostuslactone reference substance respectively appropriate, accurately weighed, add methanol and make the mixing reference substance solution of every 1mL containing andrographolide 50 μ g, dehydrorographolide 50 μ g, costunolide 100 μ g, dehydrocostuslactone 150 μ g, to obtain final product.

The preparation of need testing solution: get this product content under content uniformity item, about 3g, accurately weighed, put in tool plug conical flask, precision adds methanol 50 ml, weighed weight, supersound process (power 360 W, frequency 40 kHz) 30 minutes, let cool, more weighed weight, the weight of less loss is supplied with methanol, shake up, filter, get subsequent filtrate, to obtain final product.

Algoscopy: accurate absorption reference substance solution and each 10 μ L of need testing solution respectively, injection liquid chromatography, measures, to obtain final product.This product is containing the Radix Aucklandiae with the total amount of costunolide and dehydrocostuslactone, and every g must not be less than 2.5mg, and containing Herba Andrographis with the total amount of andrographolide and dehydrorographolide, every g must not be less than 1.0mg.

In sum, quality research have employed the index components consistent with technical study and controls, and in prescription, whole medical material all establishes the thin layer chromatography method of inspection, and Herba Andrographis, the Radix Aucklandiae, Cortex Magnoliae Officinalis three taste principal agent all establish high performance liquid chromatography content assaying method simultaneously; Above-mentioned quality research result is that the controllability of this product provides support; Results of stability confirms the stable controllability of this product.

embodiment 3 pilot process

The loose scale up test of lotus perfume is carried out according to the technique of embodiment 1, take recipe quantity 20 times of medical materials respectively, undertaken pulverizing by the loose preparation technology of lotus perfume, sieve, mix, subpackage, pick test, by production technology scale up test 10 batches of products of embodiment 1, and detect according to embodiment 2 " the loose quality standard of lotus perfume " related request, result is as shown in table 6 ~ 8.

Table 6 pilot-scale experiment

Table 7 pilot-scale experiment

Table 8 pilot-scale experiment

According to the data of above pilot process, technology controlling and process is carried out according to the scheme of pilot scale, with andrographolide, dehydrorographolide, costunolide, dehydrocostuslactone for index, tracking evaluation pilot process, result shows, production technology is smooth, simple and feasible, composition retention rate is high, and pilot product is through being up to the standards, and quality has has all met or exceeded laboratory level, explanation enlarged experiment is feasible, have operability, can instruct to amplify and produce, its cost control is suitable with ordinary powder cost.Point out this preparation technology's reasonable, be suitable for producing.

embodiment 4 pharmacodynamic experiment is studied

The present embodiment carries out the pharmacodynamic experiment research of medicine of the present invention, and all test datas adopt SAS 9.2 to carry out statistical analysis, carry out variance analysis (analysis of variance, ANOVA) or Kruskal-Wallis rank test.Data with means standard deviation ( ± SD) represent, P<0.05 is significant difference, and P<0.01 is and significant difference.

1, experiment material

(1) experimental animal: SPF level KM mice, body weight 20 ± 2g, ♀ ♂ half and half, is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center, (quality certification SCXK(Guangdong) 2013-0002).Divide cage routine to raise, raising temperature is 25 ~ 35 DEG C, and humidity is 60%, raises with solid feed, freely drinks water.

(2) test sample:

The loose sample of lotus perfume, Guangzhou Experimental Veterinary Medicine Co., Ltd. of South China Agricultural University provides, lot number: 2013030103(is according to the above-mentioned production technology preparation of the present invention).

Folium Sennae (lot number: YPA3A0001 is purchased from Guangzhou Cai Zhilin Pharmaceutical chain store)

Loperamide hydrochloride capsules, specification: 2mg, (lot number: 130219290, Xian-Janssen Pharmaceutical Ltd.)

Oleum Ricini, Tianjin good fortune chemical reagent factory in morning, lot number: 20130427

Berberine Hydrochloride Tablets, Guangdong Huanan Pharmaceutical Co., Ltd, lot number: 20131214

Escherichia coli O ₇₈, purchased from China Veterinery Drug Inspection Office.

20121228), agar powder (lot number:: 20121020) producer is Qingdao GaoKeYuan Hai Bo biotech company 20130530), brain heart infusion broth BHI(lot number nutrient broth (lot number:

(3) test sample process:

The preparation of the fragrant aproll extract of lotus: get the loose 62.5g of lotus perfume respectively, 50g, 40g, every portion adds 9,6 times amount water extraction twice respectively, extraction time is 1.5h, 1h, each time medicinal liquid filters, and is concentrated into the gradient concentration medicinal liquid making 0.625g/ml, 0.5g/ml, 0.4g/ml after merging in 85 DEG C with Rotary Evaporators, for subsequent use in 4 DEG C of Refrigerator stores.

The preparation of Folium Sennae leachate: before use, takes Folium Sennae 10g(and crosses 20 mesh sieves), with 10 times amount distilled water, soak 0.5h in 90 DEG C, get filtrate reduced in volume to 0.1g/ml medicinal liquid.

The preparation of loperamide hydrochloride solution: take loperamide hydrochloride 4mg, adds appropriate distilled water, and after ultrasonic dissolution, adding distil water is joined final concentration is 0.1mg/ml medicinal liquid, saves backup in 4 DEG C.

The preparation of berberine hydrochloride solution: get Berberine Hydrochloride Tablets 10 (altogether hydrochloric berberine 1g), add appropriate distilled water, after ultrasonic dissolution, adding distil water is joined final concentration is 1.5mg/ml medicinal liquid, saves backup in 4 DEG C.

Charcoal end suspension: take 5g active carbon, 1g sodium carboxymethyl cellulose, mix with 100ml distilled water, fully after stirring and get final product.

The preparation of BHI fluid medium: take brain heart infusion broth BHI dry powder 3.85g in conical flask, add 100mL double distilled water, and use turbula shaker shaken well, for subsequent use after autoclaving.

The preparation of BHI solid medium: take BHI dry powder 11.55g, agar powder 4.5g in conical flask, adds 300mL double distilled water turbula shaker shaken well, for subsequent use after being sub-packed in glass plate after autoclaving.

(4) instrument and equipment

Vacuum pump using circulatory water SHZ-D (III) (Yuhua Instrument Co., Ltd., Gongyi City, maximum vacuum/Mpa:0.098);

The two outstanding T1000 electronic balance (Shuan Jie test instrunment factory of Changshu City, product type T1000) of the G & G U.S.;

EYELA SB-1000 type Rotary Evaporators (Shanghai Ai Lang Instrument Ltd.);

TC-15 jacket formula heating thermostat (Haining City Xinhua District medical apparatus and instruments factory);

High-pressure steam sterilizing pan (Shanghai Sanshen Medical Instrument Co., Ltd.);

Constant incubator (pond this physics and chemistry industry Co., Ltd.);

Air bath agitator (Dong Lian electronics technology development corporation, Ltd. of Harbin City).

2, lotus perfume is loose tests the impact of Folium Sennae diarrhea mice

(1) experiment grouping: the KM mice 60 of getting 20 ± 2g, ♀ ♂ half and half, random packet is blank group, model group, positive drug group, the loose high, medium and low dosage group of lotus perfume, often organize 10.

The loose high, medium and low dosage group of lotus perfume presses the corresponding medicinal liquid of 0.2mL/10g gavage, positive drug group is with 0.2mL/10g gavage Loperamide Hydrochloride solution, blank group, model group are to equal-volume distilled water, administration 5d altogether, before modeling, fasting be can't help water and is spent the night, test the same day according to the modeling of 0.2mL/10g body weight gavage Folium Sennae extracting solution, blank group to equal-volume distilled water, after modeling 30min each treated animal give corresponding medicinal liquid.

(2) start observed and recorded after administration 1h, 4h observed by each treated animal list cage, and lower berth filter paper is done wetly just to count, and per hourly changes a filter paper.The accumulative wet just number of record, wet just level, and calculate each treated animal Scours index.

Loose stool rate: loose stool number/total just number that every animal is arranged

Scours index: loose stool rate × loose stool level

Loose stool level: the degree representing loose stool, pollute with loose stool the size deciding grade and level that paper forms stain area (polluting diameter cm), be divided into 4 grades, standard progression is as follows:

Table 9 mice loose stool level (unit: cm)

(3) experimental result is as shown in table 10

The loose impact (n=10, x ± s) on Folium Sennae Diarrhea Model mice of table 10 lotus perfume

Result shows, and the loose height of each lotus perfume, middle dosage group all can obviously reduce model mice Scours index, have obvious anti-diarrhea effect.

3, lotus perfume is loose tests the impact of Oleum Ricini diarrhea mice

Operate according to the method for above-mentioned 2nd (lotus perfume is loose tests the impact of Folium Sennae diarrhea mice), modeling reagent is to the modeling of same volume Oleum Ricini.

Lotus perfume is loose as shown in table 11 to the result of the test of Oleum Ricini diarrhea mice.

The loose impact (n=10, x ± s) on Oleum Ricini Diarrhea Model mice of table 11 lotus perfume

4, lotus perfume is loose to escherichia coli O ₇₈cause the impact of peritonitis

(1) escherichia coli O ₇₈the preparation of suspension:

Take the logarithm the escherichia coli O of trophophase ₇₈be inoculated in nutrient agar panel, put 37 DEG C of constant incubators and cultivate 24h, the single bacterium colony of picking turns in nutrient broth, cultivates 18h for 37 DEG C, measure bacterial concentration with Maxwell opacity tube in water bath with thermostatic control agitator, and is 3.0 × 10 with normal saline adjustment bacterial concentration ⁸cFU/mL.By bacterium liquid (by the weight of animals 0.01mL/g) and containing 10% BaSO ₄meat soup press 1:1 mixing, be modeling suspension.

(2) divide into groups: the KM mice 60 of getting 20 ± 2g, ♀ ♂ half and half, random packet is blank group, model group, positive drug group, the loose high, medium and low dosage group of lotus perfume, often organize 10.

The loose high, medium and low dosage group of lotus perfume press the corresponding medicinal liquid of 0.2mL/10g gavage, positive drug group with 0.2mL/10g gavage berberine hydrochloride solution, blank group, model group to equal-volume distilled water, common administration 5d.Before modeling, fasting be can't help water and is spent the night, and each group of the same day of test is according to 0.01mL/g lumbar injection escherichia coli O ₇₈suspension modeling, blank group to equal-volume normal saline, after modeling each treated animal of 30min give corresponding medicinal liquid.

(3) start each treated animal death condition of observed and recorded after administration, dead animal is dissected, and observes 1 week altogether.

(4) result is as shown in table 12.

The loose impact (n=10) on escherichia coli type peritonitis mice of table 12 lotus perfume

Result shows, and the loose height of lotus perfume, middle concentration group can reduce escherichia coli fatality rate, have good protective effect.

5, the loose impact on mice Intestinal pushing function of lotus perfume

(1) divide into groups: the KM mice 50 of getting 20 ± 2g, ♀ ♂ half and half, high, medium and low dosage group that random packet is blank group, positive drug group, lotus perfume fall apart, often organizes 10.

The loose high, medium and low dosage group of lotus perfume presses the corresponding medicinal liquid of 0.2mL/10g gavage, blank group to equal-volume distilled water, positive drug group to 0.2mL/10g gavage Loperamide Hydrochloride solution, altogether administration 5d, before modeling, fasting be can't help water and is spent the night, after test administration on same day 1h, each group of gavage gives 5% charcoal end suspension 0.2mL/10g, put to death animal after 20min, open abdominal cavity and be separated mesentery, get the intestinal tube of pylorus to ileocecus, measure total small intestinal length and charcoal end advance distance, and calculate propelling rate.

Charcoal end propelling rate=charcoal end advance distance/total small intestinal length × 100%

(2) result is as shown in table 13.

The loose impact (n=10, x ± s) on mice Intestinal pushing of table 13 lotus perfume

Note: *: compare with blank group, P < 0.05; *: compare with blank group, P < 0.01.

Result shows, and the loose height of lotus perfume, middle concentration group all obviously can suppress mice Intestinal pushing function.

6, in sum, this result of the test shows, can effectively reduce by reagent lotus perfume is loose the mortality rate that escherichia coli cause peritonitis mice; Suppress the diarrhoea of Folium Sennae and Oleum Ricini Diarrhea Model mice simultaneously effectively, be wherein excellent with high, middle dosage group, also can suppress Intestinal motility; Also for veterinary drug clinical trial provides foundation, the herbal medicine of anti-colibacillosis can be developed further.

the experimental clinical research of embodiment 5

According to " veterinary Chinese medicine, natural drug clinical trial technological guidance principle ", the loose prevention effect to test chicken artificial challenge colibacillosis of Study of Traditional Chinese Medicine lotus perfume and best dosage, for the concrete dosage regimen in Control of Colibacillosis of Chickens work common in clinical poultry provides test data and scientific basis.

1, experiment material

(1) experimental animal: 1 age in days Lingnan Yellow commercial broiler flocks, purchased from Guangdong Wiz Agricultural Science & Technology Co., Ltd..

(2) test drug: lotus perfume is loose: Guangzhou Experimental Veterinary Medicine Co., Ltd. of South China Agricultural University provides, lot number: 2013030102(is according to the above-mentioned production technology preparation of the present invention).

Four taste Herba Andrographis fall apart, and Guangzhou Experimental Veterinary Medicine Co., Ltd. of South China Agricultural University provides, lot number: 2013121301.

(3) test apparatus: constant incubator, Shanghai leap medical apparatus and instruments factory produces; Aseptic operating platform, Suzhou hundred million reaches cleaning laboratory equipment company limited and produces.

(4) strain: strains tested is chicken colibacillosis O ₇₈(purchased from China Veterinery Drug Inspection Office, lot number: CVCC1569).

(5) experimental animal basal diet: with reference to NRC(1994 version) feeding of broiler standard and China Broiler feeding standard preparation basal diet in 1998, not drug containing additive in basal diet.

2, the preparation of test organisms and counting

By chicken colibacillosis (O ₇₈serotype) inoculate ordinary flat, after 37 DEG C of cultivation 24 h, select colonies typical and be inoculated in meat soup, take out after cultivating 18 h through 37 DEG C again.By this broth culture inoculation chicken, also can by lethal for inoculation chicken in the hope of its pathogenicity rejuvenation.Inoculating slat chain conveyor from the liver separating Escherichia coli of dead chicken, finally select colonies typical ordinary broth and cultivate, adopt colony counting method to carry out colony counting, is 2 × 10 containing bacterium number by the dilution of bacterium liquid ⁹cfu/mL, for subsequent use.In the clinical symptoms of inoculation fore-and-aft observing chicken, cut open the dead chicken of inspection, get liver, spleen and heart and carry out bacteria distribution cultivation and qualification, to determine whether successfully to induce an illness.

3, the foundation of artificial challenge's chicken colibacillosis pathological model

By 0.2 × 10 ⁹cfu/mL ~ 1 × 10 ⁹bacterial concentration between cfu/mL is adjusted to 9 concentration groups, is respectively 0.2 × 10 ⁹cfu/mL, 0.3 × 10 ⁹cfu/mL, 0.4 × 10 ⁹cfu/mL, 0.5 × 10 ⁹cfu/mL, 0.6 × 10 ⁹cfu/mL, 0.7 × 10 ⁹cfu/mL, 0.8 × 10 ⁹cfu/mL, 0.9 × 10 ⁹cfu/mL and 1 × 10 ⁹cfu/mL.Often organize 20 chickling, every chicken each lumbar injection counteracting toxic substances bacterium liquid 0.5mL.Separately set one group as normal healthy controls group.The state of chickling after observation counteracting toxic substances, record chicken death number of elements.

Result of the test shows, with 0.6 × 10 ⁹cfu/ml and 0.7 × 10 ⁹cfu/ml, every chicken lumbar injection 0.5ml, test chicken 100% can be made to fall ill, present typical chicken colibacillosis clinical symptoms, fatality rate is 65%.Therefore, escherichia coli O in experiment can be inferred ₇₈strain is to the LD of chickling artificial challenge on the 10th ₅₀0.2 ~ 0.6 × 10 ⁹between cfu/ml.In formal counteracting toxic substances process, this test and Selection concentration is 0.6 × 10 ⁹the 0.6ml Escherichia coli bacteria liquid of cfu/ml is as the counteracting toxic substances dosage setting up chicken escherichia coli infection model.

4, experimental animal grouping and process

Test packet transaction situation is in table 14.By 150 1 age in days test chickens, when raising to 10 age in days, choose 120 close test chickens of healthy weight, be divided into 6 groups at random, often organize 20.1st ~ 3 groups is that test medicine group (infects, test medicine is treated), wherein the 1st group is low dosage (0.5% lotus perfume is loose) treatment group, and the 2nd group is middle dosage (1% lotus perfume is loose) treatment group, and the 3rd group is high dose (2% lotus perfume is loose) treatment group, 4th group is that control drug group (infects, to fall apart control drug to 0.5% 4 taste Herba Andrographis), the 5th group for infecting matched group (infecting, not administration), 6th group be normal healthy controls group (not infecting, not administration) and with the isolated rearing of 1-5 group.To every test chicken inoculation chicken colibacillosis (O of 1st ~ 5 groups ₇₈serotype) meat soup bacterium liquid 0.6 ml/, the 6th group of injection normal saline.Namely carry out process dispensing by table 1 after counteracting toxic substances, 1-4 group is dispensing 7 d continuously, observe 14 d to inoculation.At duration of test test chicken free choice feeding, drinking-water, and program carries out epidemic disease immunity, hygienic cleaning and sterilization routinely.

Table 14 tests grouping and disposition

5, the main Testing index of duration of test and result

Test data represents with (mean+SD), and applies SPSS statistical analysis software and carry out statistical analysis and process.

(1) the clinical growth of test chicken and onset state etc. after manual-induced broiler colibacillosis

Examine the clinical manifestation situations such as test chicken is searched for food, drunk water, diarrhoea in duration of test every morning, afternoon and do itemized record, and recording test chicken death time and quantity, pathological anatomy being carried out to death experiments chicken and bacteria distribution is cultivated.

Result of the test shows, and other 5 the infected group test chickens except the 6th group start morbidity after counteracting toxic substances 6 h, shows as lassitude, dirty and messy by hair, inverse vertical, flocks together, necking down, wing are sagging, row's HUANGBAI(sic) loose stool, drink loss of appetite.Cut open the dead chicken of inspection, observe viscera, main manifestations is perihepatitis and pericarditis, liver enlargement, hyperemia, severe patient liver surface has fibrous exudate, and air bag thickens, muddiness, Glisson's capsule thickens and has petechia or hemorrhage band, and pericardium thickens, and small intestinal mucosa has petechia.A large amount of typical escherichia coli are all isolated from liver, spleen and heart.

Result of the test shows, by tests such as test chicken clinical symptoms, bacteria distribution cultivations, determines successfully to bring out chicken colibacillosis in this test.

(2) the loose observation of curative effect to test chicken colibacillosis of lotus perfume

The recovery situation of duration of test record test chicken after morbidity after Drug therapy, record and the sickness rate, mortality rate, cure rate, effective percentage, the inefficiency that calculate test chicken.

Mortality rate: at duration of test, affected animal occurs that the classical symptom of chicken colibacillosis is also dead, and necropsy has the characteristics of lesion of typical chicken colibacillosis, and separation and Culture goes out chicken colibacillosis from liver, spleen and heart, be judged to infect death, and calculate mortality rate according to death toll gauge.

Cure rate: at duration of test, the affected animal mental status, appetite after administration recover normal, and the clinical symptoms such as excrement that no longer occur having loose bowels all belong to healing, and calculate cure rate according to healing quantity.

Effective percentage: at duration of test, affected animal its clinical symptoms after administration has obvious alleviator, is judged as effectively, and calculates effective percentage according to its quantity.

Inefficiency: at duration of test, affected animal its clinical symptoms after administration has no obvious improvement, and add severe one even to some extent, it is invalid to be judged as, and calculates effective percentage according to its quantity.

The loose therapeutic outcome to artificial challenge's chicken colibacillosis test broiler of Chinese medicine lotus perfume is as shown in Table 15.

(unit: only) added up by table 15 respectively group test chicken death time and number of elements

Result of the test shows, about 6 h after inoculation chicken colibacillosis, and clinical symptoms appears in total Test chicken, and sickness rate is 100%, and after treating 7 d through use various dose Chinese powder medicine, each test group chicken death time and number of elements are in table 15.Result shows, and 4 d have merely hit 12 and all concentrated on front 4 d test chicken death most time of concentration, particularly the 5th group of (infection matched group) test chicken death toll 13 before the test, account for more than 90% of dead sum; And 1st ~ 4 groups (infecting, treatment) account for about 70% of total death toll at the dead number of elements of front 4 d.Meanwhile, after the 8d of experimental period, all there is not dead chicken in each test group.

Concrete efficacy data statistical result is shown in table 16.

The loose curative effect to test chicken colibacillosis of table 16 lotus perfume

Note: # represents that the infection matched group test chicken oneself of not offeing medicine crosses situation to disease is resistance to.

Result of the test display (see table 8), the 6th group of (normal healthy controls group) test chicken sickness rate is for be 0% with mortality rate, and the 5th group of (infection matched group) test chicken mortality rate is 65%, apparently higher than the mortality rate (see table 3) of 1st ~ 4 groups (infecting, treatment group); Wherein, the mortality rate of the 2nd, 3 group (1% lotus perfume falls apart, 2% lotus perfume falls apart) is all lower than the 1st, 4 group (0.5% lotus perfume is loose, four taste Herba Andrographis are loose).5th group of test chicken inefficiency is all apparently higher than 1st ~ 4 groups; 5th group of (infection matched group) test chicken spontaneous recovery rate is 15%, all be starkly lower than 1st ~ 4 groups (to infect, treatment group), wherein, the cure rate of the 2nd, 3 group (1% lotus perfume falls apart, 2% lotus perfume falls apart) and effective percentage are all higher than the 1st, 4 group (0.5% lotus perfume is loose, four taste Herba Andrographis are loose).

Result of the test shows, when adding respectively in test chicken basal diet, lotus perfume is loose, four taste Herba Andrographis fall apart, and all significantly can reduce the mortality rate of artificial infection chicken's coli test chicken, and can improve its cure rate and effective percentage; Meanwhile, it is best to the curative effect of infected chicken when, 2% lotus perfume loose at interpolation 1% lotus perfume is loose.

(3) test chicken Gain weight

On-test with at the end of, to test chicken by only weighing, calculate each group of test chicken average weight.

The each group of impact of medicine on test chicken body weight is shown in table 17.

Table 17 is group test chicken body weights after off-test respectively

Note: with difference between capitalization different in string extremely significantly (p<0.01), significant difference (p<0.05) between different lower case.

Result of the test shows, when on-test, and each test group chicken average weight zero difference (96 ± 1g), find when off-test calculates survival test chicken body weight, whole infected group test chicken body weight is all remarkable in the 6th group (normal healthy controls group); Wherein, 1st ~ 4 groups of (infecting, treatment group) test chicken body weight are significantly higher than the 5th group (infection matched group).

Result of the test shows, when adding respectively in test chicken basal diet, lotus perfume is loose, four taste Herba Andrographis fall apart, and all has and promotes the resistance to mistake of artificial infection chicken coli test chicken and the effect better recovered, and can improve the upgrowth situation of test chicken; Wherein, the effect of the 2nd, 3 group (1% lotus perfume falls apart, 2% lotus perfume falls apart) is better than the 1st, 4 group (0.5% lotus perfume is loose, four taste Herba Andrographis are loose).

(4) inspection is dissected

After off-test, get each group of test chicken at random from each test group and analyse, and record relevant anatomy data.

Result of the test display, infects and the liver, heart, pericardium, small intestinal etc. of survival test chicken has no obvious pathological changes or have slight pathological changes after Drug therapy; And infect but all there is obvious pathological changes in the liver, heart, kidney, small intestinal etc. of untreated 5th group of test chicken.

Result of the test shows, when adding respectively in test chicken basal diet, lotus perfume is loose, four taste Herba Andrographis fall apart, and all can promote that artificial infection chicken's coli test chicken better recovers, and can alleviate effect of test chicken lesion tissue.

6, in this test, by manual-induced test chicken colibacillosis, and observe lotus perfume loose (high, medium and low dosage), four taste Herba Andrographis fall apart to the curative effect of infected chicken, paper examines index comprises: test chicken clinical symptoms performance situation, sickness rate, mortality rate, cure rate, effective percentage and inefficiency, and test chicken before the test after average weight, survival test chicken analyses the indexs of correlation such as pathological change.

Result of the test shows, and chicken colibacillosis is brought out in this success of the test, and morbidity chicken main manifestations is lassitude, dirty and messy by hair, inverse vertical, flocks together, necking down, wing are sagging, row's HUANGBAI(sic) loose stool, drink loss of appetite.Cut open the dead chicken of inspection, observe viscera, main manifestations is perihepatitis and pericarditis, and pericardium thickens, and small intestinal mucosa has the main pathological changes such as petechia, and all isolates a large amount of typical escherichia coli from liver, spleen and heart.

When adding respectively in test chicken basal diet, lotus perfume falls apart (high, medium and low dosage), four taste Herba Andrographis fall apart, all Chinese powder medicines all obviously can reduce the mortality rate of artificial infection chicken's coli test chicken, and its cure rate and effective percentage can be improved, alleviate test chicken main organs pathological changes, promote that test chicken resistance to mistake reaches healing with better getting well, improve the upgrowth situation of test chicken, the effect such as to put on weight.Meanwhile, comparative test result shows, is better than the loose and four taste Herba Andrographis of 0.5% lotus perfume falls apart when, 2% lotus perfume loose at interpolation 1% lotus perfume is loose to the curative effect of artificial infection chicken's coli test chicken.

In sum, this result of the test shows, when adding lotus perfume and be loose in basal diet, significantly can reduce the mortality rate of artificial infection chicken's coli test chicken, and its cure rate and effective percentage can be improved, promote the resistance to mistake of test chicken and better recovery, alleviate the lesion tissue of test chicken main organs, with the upgrowth situation improving test chicken.Comparative test result shows, when adding in basal diet that 1% lotus perfume is loose, 2% lotus perfume is loose respectively, being all better than loose and four tastes of interpolation 0.5% lotus perfume wearing lotus and fall apart the curative effect of infected chicken.

embodiment 6 expands clinical trial

According to " veterinary Chinese medicine, natural drug clinical trial technological guidance principle ", the loose prevention effect to chicken colibacillosis natural cases clinically of Study of Traditional Chinese Medicine lotus perfume and best dosage, for the concrete dosage regimen in Control of Colibacillosis of Chickens work common in clinical poultry provides test data and scientific basis.

1, experiment material

(1) experimental animal: 28 ~ 35 week age Roman egghen, Yunfu City: original breeding layer chicken company limited of Luoding City.

(3) test apparatus: constant incubator (Shanghai leap medical apparatus and instruments factory produces); Aseptic operating platform (Suzhou hundred million reaches cleaning laboratory equipment company limited and produces).

(4) experimental animal basal diet: with reference to NRC (1994) laying hen nutrition requirements preparation basal diet, not drug containing additive in daily ration.

2, the choosing and making a definite diagnosis of hens salpingitis type escherichia coli natural cases in clinical cultivation

The raw Jiang Danjiyangzhichang of original breeding layer chicken company limited in Luoding City, chooses clinical signs and goes out spiritual tired, row's brown or white fecal pollution anus, day by day become thin, egg drop reduction or stopping, producing abnormal-shape egg, the sick chicken of the clinical symptoms such as eggshell color is thin out and frangible.Carry out pathology cut open inspection visible: theca folliculi distortion, hyperemia and hemorrhage, some follicles are taupe; Tunica mucosa tubae uterinae is congested, swelling, and fallopian tube is thicker, and quality becomes fragile, rotten to the corn thinning, inside has yellow-white caseous substance and other secretions.

Carry out laboratory bacterial separation and Culture: get cheesy agglomerate pathological material of disease in fallopian tube and rule on nutrient agar panel, 37 DEG C of constant temperature culture 24 hours, present the colony growth that a kind of form is homogeneous, be characterized as circular protrusions, neat in edge, smooth moistening, semi-transparent clear and diameter 1 ~ 2 mm.Its pure culture inoculation eosin methylene blue agar is dull and stereotyped, and bacterium colony is atropurpureus, band metallic luster; Connect maconkey agar flat board, bacterium colony is shiny red.

According to above clinical symptoms, analyse pathological change and to share with laboratory bacterial and cultivate and qualification, making a definite diagnosis disease chicken is the hens salpingitis disease that escherichia coli cause.

3, experimental animal grouping and process

Test packet transaction situation is in table 18.Choose the laying hen 100 suffering from salpingitis type colibacillosis, be divided into 2 groups at random, often organize 50.1st group is the loose treatment group of lotus perfume, the 2nd group of control drug treatment group (four taste Herba Andrographis fall apart).And process by described in table 1, dispensing 7 d, observe 14 d to medication continuously.Duration of test test chicken free choice feeding, drinking-water, and program carries out epidemic disease immunity, hygienic cleaning and sterilization routinely.

Table 18 tests grouping and administrations

4, testing inspection index and result

(1) laying hen clinical manifestation after treatment by Chinese herbs is tested

Examine the clinical manifestations such as test chicken is searched for food, drunk water, diarrhoea in duration of test every morning, afternoon and do itemized record, record test chicken death time and quantity, pathological anatomy being carried out to dead chicken and bacteria distribution is cultivated.This test median oviduct scorching type colibacillosis natural occurrence laying hen respectively, four taste Herba Andrographis loose through Chinese powder medicine lotus perfume is treated 7d and observes 14d.

Result of the test shows, and along with the prolongation for the treatment of by Chinese herbs time, laying hen spirit is recovered to some extent, symptom of having loose bowels alleviates gradually, drink wish increases, and particularly egg laying performance improves gradually, and death stops gradually.

By the sick chicken of death is carried out pathology cut open inspection visible: theca folliculi distortion, hyperemia and hemorrhage, some follicles are taupe; Tunica mucosa tubae uterinae is congested, swelling, and fallopian tube is thicker, and quality becomes fragile, rotten to the corn thinning, inside has yellow-white caseous substance and other secretions.Carry out laboratory bacterial separation and Culture: get cheesy agglomerate pathological material of disease in fallopian tube and rule on nutrient agar panel simultaneously, 37 DEG C of constant temperature culture 24 hours, present the colony growth that a kind of form is homogeneous, be characterized as circular protrusions, neat in edge, smooth moistening, semi-transparent clear and diameter 1 ~ 2 mm.Its pure culture inoculation eosin methylene blue agar is dull and stereotyped, and bacterium colony is atropurpureus, band metallic luster; Connect maconkey agar flat board, bacterium colony is shiny red.According to morbidity laying hen clinical symptoms, analyse pathological change and to share with laboratory bacterial and cultivate and identify, making a definite diagnosis laying hen is the death that hens salpingitis disease that escherichia coli cause causes.

Result of the test shows, Chinese powder medicine lotus perfume is loose, four taste Herba Andrographis all have the clinical symptoms improving the hens salpingitis type disease that escherichia coli cause, and promotes that disease chicken gets well.

(2) curative effect of medication

Duration of test record morbidity test chicken at the recovery situation after Drug therapy, and calculates mortality rate, cure rate, effective percentage, inefficiency.

Mortality rate: at duration of test, there is typical Chicken salpingitis colibacillosis clinical symptoms and dead in affected animal, pathology cuts open inspection and finds: theca folliculi distortion, congested and hemorrhage, and some follicles are taupe; Tunica mucosa tubae uterinae is congested, blood stasis, and rotten to the corn thinning, quality becomes fragile, and inside has faint yellow albumen deposit; And a large amount of typical escherichia coli isolated by cheesy agglomerate pathological material of disease in fallopian tube, and calculate mortality rate according to death toll gauge.

Cure rate: at duration of test, affected animal after administration the mental status, appetite, to lay eggs etc. and all recover normal person and belong to healing, and calculate cure rate according to its quantity.

Each group of Chinese medicine to the efficacy result of morbidity test chicken as shown in table 19,20.

Respectively the group test chicken death time adds up (unit: only) to table 19

Result of the test shows, 1st, 2 test group laying hens through treatment by Chinese herbs 7d(experimental observation to treatment after 14d) after, phase (10d) has dead chicken to occur always before the test, but test chicken stops dead during 11st ~ 14d, wherein the 1st group (lotus perfume loose treatment group) at duration of test altogether dead 5 (10%), the 2nd group (four taste Herba Andrographis fall apart treatment group) at whole duration of test altogether dead 6 (12%).

Result of the test shows, salpingitis type colibacillosis natural occurrence laying hen is after treatment by Chinese herbs 7d, and the test chicken state of an illness takes a turn for the better all to some extent, dead minimizing.

The loose curative effect to test chicken colibacillosis of table 20 lotus perfume

Result of the test shows, and the 1st, 2 test group laying hens are after treatment by Chinese herbs 7d, and in the whole experimental observation phase, the loose treatment group mortality rate of lotus perfume is 10%, the four taste Herba Andrographis treatment group mortality rate that falls apart is 12%; The loose treatment group cure rate of lotus perfume is 52%, and it is 82% that cure rate adds effective percentage; The four taste Herba Andrographis treatment group cure rate that falls apart reaches 48%, and it is 76% that cure rate adds effective percentage.

Result of the test shows, salpingitis type colibacillosis natural occurrence laying hen is observing 14d through treatment by Chinese herbs 7d, and the test chicken state of an illness takes a turn for the better all to some extent, deadly reduces, part test chicken reaches criterion of cure, shows that lotus perfume is loose to the equal tool therapeutical effect of colibacillosis.

(3) test chicken laying rate and Egg Quality situation of change

Calculate each group of test laying hen egg production every day and calculate experimental period laying rate etc., calculating each group of test laying hen egg every day on average heavy, abnormal-shape egg rate, soft shell check rate etc. simultaneously.

Result is shown in table 21.

Table 21 Chinese medicine is on the impact of test chicken laying rate and Egg Quality

Note: result of the test is divided into 1 ~ 7d, 8 ~ 14d, two experimental stages are added up.

Result of the test shows, and test group laying hen is adding up the stage through treatment by Chinese herbs 7d as one, and after drug withdrawal, 8 ~ 14 d are re-used as second statistics stage, and detailed results is in table 12.Statistical result showed, 1st, 2 test group laying hens at the egg productive rate in 8 ~ 14 d stages all apparently higher than 1st ~ 7d, egg is on average heavy also a little more than 1st ~ 7 d simultaneously, and the soft shell shell-breaking rate of egg and abnormal rate are all lower than the 1st ~ 7 d, comparative test result shows, and the 1st group of (the loose treatment group of lotus perfume) resultant effect is better than the 2nd group (four taste Herba Andrographis fall apart).

Result of the test shows, salpingitis type colibacillosis natural occurrence laying hen is observing 14d through treatment by Chinese herbs 7d, the laying rate of test chicken has obvious rising, and Egg Quality also makes moderate progress, and shows that two Chinese powder medicines all have treatment and the effect improving hens salpingitis disease.

5, this clinical expanding test place is selected in the raw Jiang Danjiyangzhichang of original breeding layer chicken company limited of Yunfu, Guangdong Province Luoding City, because Yunfu, Guangdong Province Luoding City is located in more hot place, South China of China, day-light saving time is long, especially the bacterial disease that annual 6 ~ JIUYUE (particularly 7 ~ August) often occurs at feeding process, the benefit that the hens salpingitis disease particularly caused by escherichia coli affects cultivation for a long time always improves.

This test is selected in July, 2014 to August, belongs to the hens salpingitis disease period occurred frequently that being caused by escherichia coli appears in breeding layer chicken field in one-year age.In this test, by morbidity laying hen clinical symptoms, typically analyse pathological change and the method such as bacteria distribution cultivation and qualification, filter out 100 hens salpingitis disease natural occurrence laying hens caused by escherichia coli, and be divided into 2 groups at random, the 1st group (lotus perfume is loose) often organizes 50 test chickens with the 2nd group (four taste Herba Andrographis fall apart).By Drug therapy 7d, and observe 14d, paper examines index comprises: the performance of laying hen clinical symptoms, mortality rate, cure rate and effective percentage, and the index such as the laying rate of test chicken, Egg Quality.

Result of the test shows, salpingitis type colibacillosis natural occurrence laying hen is at, four taste Herba Andrographis loose treatment 7d loose through Chinese medicine lotus perfume respectively and observe 14d, along with the prolongation for the treatment of by Chinese herbs time, natural occurrence test laying hen spirit is recovered gradually, symptom of having loose bowels constantly alleviates, drink for increasing, death stops gradually, laying rate improves constantly, Egg Quality makes moderate progress, and all have about 50% laying hen to be cured in the off-test time, comprehensive evaluation result shows, and the loose four taste Herba Andrographis that are better than of Chinese medicine lotus perfume fall apart.

In sum, this result of the test shows, in South Chinas such as Yunfus, in laying hens essential ration, add that lotus perfume is loose, four taste Herba Andrographis all have the clinical symptoms improving hens salpingitis disease (being caused by escherichia coli), reduce laying hen mortality rate, improve laying rate of laying hen and improve egg quality, promoting the resistance to mistake of disease chicken and gradually recover one's health and reach criterion of cure, comprehensive evaluation result shows, and the loose four taste Herba Andrographis that are better than of Chinese medicine lotus perfume fall apart.

In addition, target animals safety test, experimental clinical trial and expansion clinical trial all result show, in basal diet, adding tested medicine lotus perfume loose, is safe and effective for preventing and treating chicken colibacillosis.Quantity is recommended to be add the tested medicine lotus perfume of 1000 g in every 100 kg diet to fall apart, continuous use 7 d.

comparative example

1, the loose prescription of lotus perfume is selected

(1) test according to the method for embodiment 4, the prescription loose to lotus perfume is selected:

Contrast groups 1: Herba Andrographis.

Contrast groups 2: Herba Andrographis, Herba Diclipterae Chinensis.

(2) detect according to the index of embodiment 4, Comparative result shows, and contrast groups 1 and 2 all has the colibacillary effect of certain suppression, can have certain therapeutic effect to escherichia coli type peritonitis mice; But compared with formula of the present invention (: Herba Andrographis 16.7%, Rhizoma Atractylodis 25%, the Radix Aucklandiae 16.7%, Cortex Magnoliae Officinalis 16.6%, Rhizoma Zingiberis 25%), the formula of contrast groups 1 and 2 to the preventive effect of escherichia coli type peritonitis significantly lower than formula of the present invention.

And the formula of contrast groups 1 and 2 does not have obvious curative effect to the diarrhoea that Oleum Ricini and Folium Sennae cause, and does not significantly advance the effect of mice intestinal function yet.In real work, chicken colibacillosis and treatment time by being permitted multifactorial impact, be not that simple suppression escherichia coli just can perfect realization.Therefore, consider each conditions, formula of the present invention (: Herba Andrographis 16.7%, Rhizoma Atractylodis 25%, the Radix Aucklandiae 16.7%, Cortex Magnoliae Officinalis 16.6%, Rhizoma Zingiberis 25%) be the Chinese medicine formula for the treatment of colibacillosis the best.

2, the loose prescription proportioning of lotus perfume is selected

Test according to the method for embodiment 4 and index, the proportioning of the loose prescription of lotus perfume is optimized, contrast through experimental result, consider each side factor, when the percentage composition of each component is following proportioning in herbal medicine compound preparation of the present invention: 5 ~ 55% Herba Andrographis, 10 ~ 65% Rhizoma Atractylodis, 5 ~ 55% Radix Aucklandiae, 5 ~ 55% Cortex Magnoliae Officinalis, 10 ~ 65 % Rhizoma Zingiberiss, the effect of control chicken colibacillosis preferably can be reached.And optimum proportioning is: Herba Andrographis 16.7%, Rhizoma Atractylodis 25%, the Radix Aucklandiae 16.7%, Cortex Magnoliae Officinalis 16.6%, Rhizoma Zingiberis 25%, and medication optimal dose is 1%.

3, the investigation of the loose area of application of lotus perfume

During in order to investigate practical application, the regional suitability that lotus perfume of the present invention is loose, we also have also been made clinical research experiment in chicken farm, Henan area, concrete grammar is with embodiment 6.

Investigation result shows, and with embodiment 6 compared with the application result of chicken farm, Luoding City, when this chicken farm application in Henan, the loose therapeutic effect for chicken colibacillosis of lotus perfume has obvious reduction, and we analyze this may be because the difference of north-south climate causes.

But with above-mentioned contrast groups 1(Herba Andrographis) with contrast groups 2(Herba Andrographis, Herba Diclipterae Chinensis) curative effect compare, when this chicken farm application in Henan, the loose therapeutic effect for chicken colibacillosis of lotus perfume still has certain advantage.Therefore, in general, lotus perfume of the present invention is loose has significant curative effect for chicken colibacillosis, is particularly useful for the subtropical climate areas such as our southern area.

Claims

1. treat the application of herbal medicine compound preparation in the medicine of preparation treatment chicken colibacillosis of chicken colibacillosis for one kind, it is characterized in that, described herbal medicine compound preparation is made up of the component of following percentage composition: 5 ~ 55% Herba Andrographis, 10 ~ 65% Rhizoma Atractylodis, 5 ~ 55% Radix Aucklandiae, 5 ~ 55% Cortex Magnoliae Officinalis, 10 ~ 65 % Rhizoma Zingiberiss.

2. applying according to claim 1, it is characterized in that, is the application in the medicine of preparation treatment subtropical climate area chicken colibacillosis.

3. apply according to claim 2, it is characterized in that, described subtropical climate area refers to South China of China.

4. treat the application of herbal medicine compound preparation in the medicine of preparation treatment chicken diarrhea of chicken colibacillosis described in claim 1, it is characterized in that, described chicken diarrhea is the diarrhoea caused by Oleum Ricini or Folium Sennae.

5. described in claim 1, treat the application of herbal medicine compound preparation in preparation suppression chicken or mice Intestinal pushing function medicament of chicken colibacillosis.

6. apply according to claim 1, it is characterized in that, the herbal medicine compound preparation of described treatment chicken colibacillosis is made up of the component of following percentage composition: 10 ~ 40% Herba Andrographis, 15 ~ 55% Rhizoma Atractylodis, 10 ~ 40% Radix Aucklandiae, 10 ~ 40% Cortex Magnoliae Officinalis, 15 ~ 55% Rhizoma Zingiberiss.

7. apply according to claim 1, it is characterized in that, the herbal medicine compound preparation of described treatment chicken colibacillosis is made up of the component of following percentage composition: 10 ~ 25% Herba Andrographis, 20 ~ 35% Rhizoma Atractylodis, 10 ~ 25% Radix Aucklandiae, 1 ~ 25% Cortex Magnoliae Officinalis, 20 ~ 35% Rhizoma Zingiberiss.

8. apply according to claim 1, it is characterized in that, the herbal medicine compound preparation of described treatment chicken colibacillosis is made up of the component of following percentage composition: Herba Andrographis 16.7%, Rhizoma Atractylodis 25%, the Radix Aucklandiae 16.7%, Cortex Magnoliae Officinalis 16.6%, Rhizoma Zingiberis 25%.

9. apply according to claim 1, it is characterized in that, the dosage form of the herbal medicine compound preparation of described treatment chicken colibacillosis is powder, preparation technology is as follows: after five kinds of Chinese crude drugs are broken into coarse granule respectively, in the mixing of prescription ratio, further fine powder is broken, crosses 30 mesh sieves, put in V-Mixer and mix 30 minutes, subpackage.