| In recent years, with the development of frog breeding industry, some of diseases began to frequent outbreaks on frogs. cataract disease is one of them, The disease has some features of spread fast,death rate,easy to repeate,causing huge economic losses to the aquaculture industry of frogs. The traditional control methods simple and crude. Having the problems of drug residue and environmental pollution. This study purified pathogen was isolated from the parts of heart and liver of diseased nigromaculata. through the artificial regression infection experiments, the test of Biochemical identify,drug allergy testing, phylogenetic analysis based on 16 S rDNA sequencing results, final confirmation of the pathogenic bacteria as Elizabethkingia menin-gosepticum, Antibiotic susceptibility tests showed Elizabethkingia menin-gosepticum is sensitive On six kinds of drug in 30 kinds of drugs, accounting for 20% of the total, it shows that it has a strong resistance, Therefore, to the healthy development of frogs aquaculture industry must seek the way that more efficient and environmentally friendly. Natural herbal medicine is an important part of traditional medicine, with the characteristics of abundant resources, significant efficacy, renewable, less pressure on the environment, Through the research of antibacterial on 60 kinds of traditional Chinese medicine, hold 60 kinds of traditional Chinese medicine into six groups, Screened out the best inhibitory effect of five kinds of Chinese medicine, Traditional Chinese Medicines subject to multiple effects, especially in the extraction process is often affected by the soaking time, ethanol concentration, extraction time, liquid ratio, extraction temperature. This study examined the effect of extract concentration on thetraditional Chinese medicine of inhibitory effect, The results show that when the low concentrations inhibitory effect with increasing concentration of the drug solution the efficacy has been enhanced, when the liquid concentration reaches a certain level, continue to increase the liquid concentration, antibacterial effect does not significantly increase.Experimental study on the single factor of five kinds of Chinese medicine found soaking time has a very significant impact on where the four kinds of traditional Chinese medicine, They are schisandra, myrobalan, barbata, gall, effect of ebony was not significant, soaking time in the range of 0 min-40 min, Inhibition zone diameter of these five kinds of Chinese medicine expanded with longer soaking time. Ethanol concentration has significant effects on two kinds of Chinese medicine, are barbata, gall, on Schisandra, myrobalan, ebony was not significant, The curve of three medicine with the changes of ethanol concentration modest, extraction time has a very significant impact on the four kinds of traditional Chinese medicine, they Are schisandra, barbata, ebony, gall, Influence of coesite was not significant, The Antibacterial effect of ebony, barbata, Schisandra are gradually increased when extraction time is within the range of 10 min-50 min, two other medicine declining or unchanged trends. Liquid-solid ratio has a significant impact on the five kinds of traditional Chinese medicine, The best inhibitory effect of 5 TCMs appears in the range of 50: 1 to 60: 1, Effect of extraction temperature are significant on five Chinese herbs, The inhibitory effect of 5 kinds of Chinese medicine uptrend when the temperature increased from 30 ℃ to 90 ℃ in the process, the smallest rise is coesite, the efficacy of the other four kinds of traditional Chinese medicine of the magnitude are greatly improve, by this experiment to determine the main factors influencing the efficacy of each extract of herbs, it provides the basis data for the extraction technology of Chinese medicine.Through preliminary experiments to determine the main factors affect the efficacy of traditional Chinese medicine, depending on the circumstances of each medicine designed five corresponding orthogonal experiment. Study the extraction time,liquid ratio, extraction temperature,in the orthogonal experiment of ebony, they were set up three levels, Experimental results show that the liquid antibacterial effect improved 7.81% in the use of optimized extraction process, The orthogonal experiment of schisandra investigated the soaking time, extraction time, liquid ratio, extraction temperature, set three levels, Verification experimental results show that extraction technology to effectively improve 6.83% of the efficacy of schisandra, it is to improve the efficacy of the smallest. Gallic orthogonal experiments involving four factors, are soaking time, extraction time, liquid ratio, extraction temperature, experimental effectively improve the efficacy of gallic 13.13%. is the sharpest increase in the five kinds of Chinese medicine. coesite orthogonal experimental design three factors,them are soaking time, liquid ratio, extraction temperature, set three levels, Enhance the antibacterial effect 10.79% of coesite, Banzhilian orthogonal experiment investigated five factors, set three levels, optimization process to improve the inhibitory effect of 10.71%,Comparison the Differences between Optimization and Common by ANOVA.,the results show that the difference is significant, description Optimization of extraction process of each medicine can significantly improve the inhibitory effect of the five kinds of Chinese medicine.Orthogonal experiment to optimize the extraction process of each medicine, for the study of Compound provides important theoretical basis. put The five kinds of traditional Chinese medicine combined into 10 Compound by checkerboard method in the Experiment study of Compoun. first, do these 10 kinds of compound synergy study, according to the method of interpretation that If DHC≤0.5 as a significant synergistic effect, 0.5 <DHC <1 synergistic effect, 1≤DHC <2 is irrelevant action, 2≤DHC antagonistic, interpretation of the synergistic effect of compound, The results showed significant synergy, synergy unrelated action, each 3 Compound, antagonism has a compound, the further research of the Compound proportion results show that play a leading role is more potent single herb in Compound, Get a compound Herb is A2D8 that the best inhibitoryeffect.Through the above study found pathogens that cause frogs cataracts disease, Found the medicine that fight the pathogen and single herb, optimize the extraction conditions of single herb, screened out the better compound Chinese medicine, for the use of Chinese herbal medicine provides a theoretical basis in the prevention and treatment of cataract disease in frogs. |
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