| Fowl cholera is a kind of poultry diseases and contagious diseases caused by pasteurella multocida,which are mainly harmful to poultry and wild birds, also known as pasteurella diseases or poultry hemorrhagic septicaemia. Naatural incubation period is generally 2-9 d, it’s often present symptoms of septic.morbidity and mortality rate is very high, but sometimes changing into chronic or benign. Fowl cholera treatment are mainly composed of western medicine, although have a certain therapeutic effect, but western medicine viscera poisonous side effects of poultry, and problems existing antibiotics residue in the body. Natural Chinese medicine is to avoid the harm the animals themselves, and not cause drug residues. Traditional Chinese medicine do no harmful to the animals, and cause not drug residues. Out a few of the experiment of TCM from 20 strong antimicrobial activity of traditional Chinese medicine, and conduct joint bacteriostatic experiment, selected with additive and synergistic action of traditional Chinese medicine formula used for clinical curative effect observation and acute toxicity experiment. The main content results are as follows:1. Extraction of active ingredients of 20 kinds of traditional Chinese medicine were in vitro antimicrobial activity to Psteurella multocida. By 20 herbs using ultrasound, ethanol reflux, water bath heating method of extracting the active ingredients, the active ingredient in traditional Chinese medicine extracted from in vitro inhibition of Pasteurella multocida test, measure the active ingredients of traditional Chinese medicine for Pasteurella multocida minimum inhibitory concentration (Minimal inhibitory concentration, MIC) and minimum bactericidal concentration (Minimal bactericidal concentration, MBC), and the active ingredients of traditional Chinese medicine combined inhibition was determined by checkerboard method for Pasteurella multocida effect. The results showed that:With the result the minimal inhibitory concentration (MIC) was in a range of 6.25-50 mg/mL for Moxa leaf, Rhizoma coptidis, Cortex phellodendri, Cortex fraxini, Garden burnet, Giant knotweed’s extracts Moxa leaf, Rhizoma coptidis, Cortex phellodendri, Cortex fraxini, Garden burnet, Giant knotweed’s extracts; MIC was in range of 50 to 100 mg/mL for Radix paeoniae rubra, Radix-polygoni multiflori, Herba artemisiae capillaris, Rhizoma belamcandae, Licorice root. Dandelion, Cortex dictamni’s extracts; Flos lonicerae, Radix bupleuri, Radix isatidis, Selfheal, Andrographis, Rhizoma’s extracts of MIC are more than 100 mg/mL. The results of joint bacteriostatic test showed that the fractional inhibitory concentration index (FJCI) of Rhizoma coptidis, Cortex phellodendri, Cortex fraxini, Giant knotweed, and Artemisia argyi were less than or equal to 0.5 and the fractional inhibitory concentration index (FICI) of Cortex phellodendri, Cortex fraxini Rhizoma coptidis and Giant knotweed; Qin leather, Garden burnet and Cortex phellodendri were more than or equal to 2. So we could have a conclusion that Rhizoma coptidis, Cortex phellodendri, Garden burnet. Folium artemisiae argyi have good antimicrobial activities against pasteurella multocida; Rhizoma coptidis. Cortex phellodendri, Garden burnet and Mugwort showed additive or synergy action; Sanguisorba and Cortex phellodendri were showed antagonistic action. Finally selected rhizoma coptidis, cortex phellodendri and mugwort the three traditional Chinese medicine formula.2.Optimum research formula granules. The aim was to screen the best extraction technology of anti Pasteurella activity of three traditional Chinese Medicine such as Artemisia argyi effective parts.The extract yield of effective parts,minimal inhibitory concentration (MIC) of extracts to Pasteurella and minimum bactericidal concentration (MBC) of extacts to Pasteurella were used as three indicators comprehensive evalu-ation.The best extraction technology was screened by orthogonal design with five factors, which were solid-liquid ratio, extraction time, each time of extraction, concentration of ethanol and temperature. Artemisia argyi:50% ethanol, 70℃,2 hours every extracting time,30 times the solvent; Rhizoma coptidis,70% ethanol,12 times the solvent, extracting 2 times,1 hour every time;Cortex phellodendri,70% ethanol,7 times the solvent, extracting 3 times,80 minutes every time.3. Acute toxicity experiment of formula granules and the artificial infection curative effect observation of Pasteurella multocida. According to the veterinary drug research technical guidelines for traditional Chinese medicine formula extract lavage acute experiment was carried out. Drug doses more than 5000 mg/kg, if rats without any reaction and without death, LDso> 5000mg/kg. Then Using mice to Pasteurella multocida building, using traditional Chinese medicine formula granules to model mice of high, medium and low (780 mg/ml,390 mg/ml,78 mg/ml) treatment dose groups According to body weight in mice every 10 g to 0.1 ml. High dose group of survival rate was 80%; Dose group survival rate was 80%; Low dose group survival rate was 50%; The control group survival rate was 20%. Conclusion:Chinese medicine formula was non-toxic. High, middle dose groups have certain protective effects on the animals infected with Pasteurella multocida providing a theoretical basis for clinical drug development.4.The preparation of formula granules and its quality control. By orthogonal design method of the formula granules material:alpha, lactose and soluble starch and dextrin filtered according to the different proportion, and Using HPLC and ultraviolet spectrophotometer on berberine in granules and eucalyptol content were determined. The results showed that: Traditional Chinese medicine formula solution with the best ratio of excipients forl:5, The chromatographic conditions are acetonitrile and Potassium dihydrogen phosphate(30:70,v/v); Column temperature is 30℃;speed is 1.0 mL℃in-1;The wavelength is 265nm; Sample quantity is 20μL, berberine content in the formula granules is about 18.67ug.The conclusion:according to orthogonal test screening of excipients granules size, water content, clarity all conform to the requirements of the pharmacopoeia, its quality standards within the prescribed scope. |
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