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机构地区:[1]四川大学华西基础医学与法医学院,成都610041 [2]西南大学生物技术中心,重庆400716
出 处:《四川大学学报(自然科学版)》2007年第5期1091-1096,共6页Journal of Sichuan University(Natural Science Edition)
摘 要:小皱蝽是半翅目蝽科外生翅类昆虫,以其为实验材料,蘸取大肠杆菌和金黄色葡萄球菌菌液,针刺小皱蝽成虫腹部,诱导免疫防御反应,24 h后收集虫体.应用11种溶剂提取昆虫抗菌物质,黑曲霉生长抑制试验及琼脂孔半径扩散试验检测提取物的抗真菌、抗细菌活性,筛选最适提取溶剂.结果表明,80%甲醇能较好提取小皱蝽成虫的抗真菌、抗细菌物质,0.05%TFA的昆虫生理盐水浸提能较好提取小皱蝽成虫的抗细菌物质.两种抗菌物质提取液经超速离心、超滤、固相萃取分级分离,活性回收率分别为14.22%、11.54%,纯化倍数分别为4.64、5.36.本实验为昆虫生物活性物质的筛选和制备提供实验依据.Cyclopelta parva Distant adults, an hemipteran insect with an incomplete metamorphosis was one of endopterygote insects, which was pricked in abdomen with needle dipped into a bacterial combined pellet of Escherichia coli D22 and Staphyloccus aureus, thus the wound infection and immunal defense response were induced by the septic injury, and extracting the antimicrobial substances by eleven extraction solutions. The antifungal activity was detected by the growth inhibition of Aspergillus niger in agar plate and the antibacterial activity was analysed by the radial diffusion assay. The optimal solvents were exploited to extract the an- timicrobial substances. The antimicrobial activity of the extracted solutions were detected by the radial diffusion assay in agar pore. The results showed that 80 % methanol could extract the antifungal and antibacterial substances in C. parva Distant adults, and insect physiological saline of 0.05 %TFA could extract antibacterial substances in C. parva Distant. Both of the extraction solutions were fractionated by ultracentrifugation, ultrafiltration and solid phase extraction, and the antimicrobial activity were analysed. Purification folds were 4.64 and 5.36 and the activity recovery were 14.22 % and 11.54 % respectively.
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