中草药有效成分酶解法提取工艺优化及其不同配方对引起仔猪腹泻细菌的抑制作用  

Optimization of Enzymolysis Method Extracting Active Ingredients from Chinese Herbs and Inhibitory Effect of Their Different Formulas on Bacteria Causing Diarrhea of Piglets

作　　者：李文星 武迪 孙毅 曲威[1] LI Wenxing;WU Di;SUN Yi;QU Wei(Yantai Research Institute of China Agricultural University,Yantai,Shandong 264670,China)

机构地区：[1]中国农业大学烟台研究院,山东烟台264670

出　　处：《贵州农业科学》2023年第12期75-83,共9页Guizhou Agricultural Sciences

基　　金：烟台市校地融合发展项目“基于生物技术的断奶仔猪应激综合症的绿色防治模式”(2021XDRHXMQT19);中国农业大学烟台研究院“一流教研室”建设项目(202201Ys)。

摘　　要：【目的】探明中草药有效活性成分的提取条件,并筛选抑制仔猪腹泻细菌的高效中药方剂,为生猪养殖过程中绿色防治断奶仔猪腹泻提供理论依据。【方法】采用酶解法提取中草药的有效成分,通过单因素试验研究有效成分提取的温度、时间、pH、酶用量、料液比和处理方式对提取效果的影响,从而确定酶解的最佳条件;利用抑菌圈法测定不同中草药对肠产毒性大肠埃希氏菌、猪霍乱沙门氏菌和绿脓杆菌的抑菌情况,筛选具有较好抑菌效果的中药方剂。【结果】酶解法提取中草药有效成分的最佳工艺为提取温度55℃+提取时间2 h+酶添加量为3%+料液比为1∶25,pH 10,提取前对中草药进行研磨处理。肠产毒性大肠埃希氏菌对单味中草药连翘表现极敏〔抑菌圈直径(Φ)22.2 mm〕,对石榴皮(Φ15.7 mm)和山楂(Φ19.4 mm)表现高敏,优化配方为山楂+虎杖+石榴皮+苦参(Φ18.4 mm);猪霍乱沙门氏菌对单味中草药黄芪(Φ19.2 mm)和川黄柏(Φ15.5 mm)表现高敏,优化配方为黄芪+川黄柏+石榴皮(Φ24.4 mm);绿脓杆菌对单味中草药板蓝根(Φ11.5 mm)和党参(Φ12.1 mm)表现高敏,优化配方为党参+鱼腥草+甘草+龙胆草(Φ15.0 mm)。【结论】中药方剂山楂+虎杖+石榴皮+苦参、黄芪+川黄柏+石榴皮和党参+鱼腥草+甘草+龙胆草分别对引起仔猪腹泻的肠产毒性大肠埃希氏菌、猪霍乱沙门氏菌和绿脓杆菌具有高效抑制作用。【Objective】The extraction conditions of effective active components of Chinese herbs were explored,and the efficient Chinese herbal formulas to inhibit the bacteria causing piglet diarrhea were screened,which provided the theoretical basis for preventing and controlling diarrhea of weaned piglets in the process of pig breeding.【Method】The active ingredients in Chinese herbs were extracted by the enzymolysis method,and the effects of temperature,time,pH,enzyme dosage,material-liquid ratio and treatment on the extraction of the active ingredients were investigated through a single factor test to determine the optimal conditions for the enzyme extraction;On this basis,the bacteriostatic circle method was used to determine the bacteriostatic situation of different Chinese herbs against enterotoxigenic Escherichia coli,Salmonella choleraesuis and Pseudomonas aeruginosa,and then Chinese herbal formulas with better bacteriostatic effects were screened out.【Result】The optimal process for the enzymolysis method extracting the active ingredients of the Chinese herbs included 55℃of extraction temperature+2 h of extraction time+3%enzyme amount+1∶25 of material-liquid ratio,pH 10 and grounding the Chinese herbs before extraction.Enterotoxigenic E.coli was extremely sensitive to the single herb Forsythia suspensa〔(Φ)22.2 mm of inhibition zone diameter〕and highly sensitive to pomegranate peel(Φ15.7 mm)and hawthorn(Φ19.4 mm),and the optimized formula was hawthorn+Polygonum cuspidatum+pomegranate peel+kuh-seng(Φ18.4 mm);S.choleraesuis showed high sensitivity to the single herb Astragalus membranaceus(Φ19.2 mm)and Phellodendron chinense(Φ15.5 mm),and the optimized formula was A.membranaceus+P.chinense+pomegranate peel(Φ24.4 mm).P.aeruginosa was highly sensitive to the single herb Radix isatidis(Φ11.5 mm)and Codonopsis pilosula(Φ12.1 mm),and the optimized formula was C.pilosula+Houttuynia cordata+Glycyrrhiza uralensis+Gentiana cruciata(Φ15.0 mm).【Conclusion】The Chinese herbal formulas of hawthorn+P

关 键 词：中草药 酶解法 提取工艺 抑菌率 仔猪腹泻 肠产毒性大肠埃希氏菌 猪霍乱沙门氏菌 绿脓杆菌 

分 类 号：S858.28[农业科学—临床兽医学]