复方中药提取工艺优化及抗致羔羊腹泻病原菌作用研究

Study on Extraction Process Optimization of Compound Traditional Chinese Medicine and Its Anti-Pathogenic Bacteria Causing Diarrhea in Lambs

试验旨在优化复方中药最佳提取工艺,探究复方中药抗致羊腹泻病原菌的作用。试验分离并鉴定了致羔羊腹泻病原菌,采用多孔板-噻唑蓝(MTT)比色法测定复方中药提取液对病原菌的抑制活性;然后,根据中药药性,筛选出甘草、秦皮、干姜等多味中药,基于单因素和正交设计优化提取工艺,以对病原菌的抑制率为评价指标,研究了乙醇浓度、料液比、回流时间等3个因素对复方中药提取工艺的影响;确定最佳提取工艺后,进行了中药有效成分抑菌试验及临床抗腹泻试验。结果显示:复方中药的最佳提取工艺参数为:乙醇浓度30%、料液比1∶10 g/L、回流时间2 h,该条件下提取物对致羊腹泻的大肠杆菌和沙门氏菌均具有明显的抑制作用,抑制率分别为74.68%、78.16%,平均抑制率为76.42%;腹泻羔羊饲粮中添加适量复方中药提取物7d后,其腹泻指数极显著降低(P<0.01)。试验为复方中药在治疗羊腹泻疾病中的应用提供理论参考。

In order to optimize the extraction process of compound Chinese medicine and explore the effect of compound Chinese medicine on pathogenic bacteria of sheep diarrhea.The experiment first isolated and identified the pathogenic bacteria causing sheep diarrhea.The inhibitory activity of the extract against the pathogenic bacteria was determined by using the porous plate-thiazole blue(MTT)colorimetry.Based on the properties of the traditional Chinese medicine,multiple traditional Chinese medicines such as licorice,Qinpi,and dried ginger were selected.The extraction process was optimized based on single factor and orthogonal design.The effects of ethanol concentration,solid-liquid ratio and reflux time on the extraction process of compound traditional Chinese medicine were studied.The bacteriostasis test and clinical anti-diarrhea test of active components of Chinese herbal medicine were carried out after the optimal extraction process was determined.The results showed that the extract of the compound Chinese medicine had obvious inhibitory effect on Escherichia coli and Salmonella causing diarrhea in sheep.The best extraction process of the compound Chinese medicine was scientifically optimized.The parameters were ethanol concentration of 30%,solid-liquid ratio of 1∶10,and reflux time of 2 h.Under this condition,the inhibition rates of Enterobacter coli and Salmonella were 74.68%and 78.16%,respectively,with an average inhibition rate of 76.42%.The clinical trial results showed that aftter adding an appropriate amount of compound Chinese medicine extract to the diet of diarrhea lambs for 7 days,the diarrhea index singnificantly decreased(P<0.01).The experiment provides theoretical reference for the application of compound Chinese medicine in the treatment of sheep diarrhea disease.