Analysis Of The Active Ingredients Of Huang Lian Extracts And Preliminary Research On Outer Membrane Protein Of Riemerella Anatipestifer

Riemerella anatipestifer disease is an infectious disease that threats to the duck industry seriously, which is caused by Riemerella anatipestifer . There is no effective prevention methods except for the antibiotic therapy. But the RA become resistant to antibiotics easily. Huang Lian Tang,a new kind decoction of pure Chinese herbs, was developed by my instructor ducks at the Zhejiang Academy of Agricultural Sciences, which is extracted from 3 kinds of herbs of Coptis chinensis Phellodendron amurense and Glycyrrhiza uralensis, The inhibition rate of Huang Lian Tang to the epidemic Riemerella anatipestifer disease is over 98%. The extract method has been applied for patent. In order to understand the mechanism of Huang Lian Tang and to provide the theoretical basis for the exploiture veterinary medicine to prevent and cure RA disease preliminary study on the content of effective active ingredient, inhibitory rates and effect on outer membrane protein of huang lian tang has been carried out in this paper.The effective component are extracted by Water Extraction and Alcohol Precipitation. The 6 major components are determined and their contents are tested with high performance liquid chromatography. The content of each component is as follows:Berberine Hydrochloride 5.7 mg·g^-1,Epiberberine 0.29973 mg·g^-1,Jatrorrhizine 0.10866 mg·g^-1,Palmatine 0. 254 mg·g^-1 and Glyeyrrhizinate 0.009 mg·g^-1,Glycerrhetinic acid 0.000 mg·g^-1 were contained in huang lian tang respectively. The determination of Huang lian tang main components lays foundation for further pharmacological experiments and standardization of Huang Lian Tang.Inhibitory effect on RA of huang lian tang is researched by Vitro antibacterial method. Standard concentration curve of RA was drafted at first, and then bacteria of 20μL, 50μL, 120μL,190μL and 250μL are added to medium with different drug concentrations and the UV absorbance are tested after 12 h culture in .The bacterial content of various medical concentrations were calculated and then the inhibitory rate of the bacteria were measured, and inhibition rate of analysis of variance were carried on at last. The results show that the inhibitory effect of the medicine is significantly, The study of medicine lay foundation of extracting outer membrane protein under the effects of the medicine at the next step.outer membrane protein, a transmembrane protein attached to the bacterial cell membrane, is one of the factors related to bacteria infection. The comparation of Electrophoretic band of bacteria with medicine and without out medicine is carried out in SDS-PAGE experiment. The results of the blank group bacteria Omp of serotype 1 in SDS-PAGE experiment showed that it was composed of 12 to 15 protein polypeptides, and the molecular weight of the bands are from 20 to 100 ku. But the electrophoretic bands of RA with medicine disappeared partly or thoroughly. Inhibitory effect of huang lian tang on outer membrane protein was clarified initially, Hence Mechanism of Therapy of huang lian tang is revealed at another level .