摘要
为探索金属离子螯合剂乙二胺四乙酸(EDTA)单独及联合抗菌药物对水霉菌生物膜的清除效应,采用微量稀释法测定EDTA对水霉菌游动孢子最小抑菌浓度(MIC)。用常用剂量和2倍剂量抗菌药亚甲基蓝、美婷及1、4、16倍MIC的EDTA联合二者常用剂量分别作用于水霉菌生物膜,CCK-8法检测药物作用后生物膜活力变化。并用Calcofluor White M2R染色EDTA处理后水霉菌生物膜,荧光显微镜观察胞外基质变化。结果显示,亚甲基蓝、美婷在常用剂量下作用于水霉菌生物膜效果不佳,与对照组无显著性差异(P>0.05),亚甲基蓝在2倍剂量下仍不能发挥较好的效果,而亚甲基蓝、美婷与EDTA联合作用均表现出较好的作用。EDTA作用后水霉菌生物膜结构破坏,胞外基质减少。研究结果表明,EDTA可破坏水霉菌生物膜结构,与抗菌药联合使用对生物膜有显著的清除作用。
The clean-up effect of EDTA,a metal-chelator,was investigated when used alone or combined with antimicrobial agents on Saprolegnia biofilm.The minimal inhibitory concentration (MIC)of EDTA to Saprolegnia spore was measured by microdilution broth method.Saprolegnia biofilms were treated by common and 2 times dose of anti-Saprolegnia drugs.Meanwhile,Saprolegnia biofilms were treated by 1 MIC,4 MIC and 16 MIC EDTA combining with common dose of methylene blue and Mei Ting,respective-ly.And then all results were measured by the CCK-8 (Cell Counting Kit-8).Furthermore,the biofilm treated by EDTA was stained with calcofluor white M2R,observed by fluorescence microscope,to investi-gate the change of the extracellular maxtrix.The results showed that,compared with the control group,it played no significant role when the biofilms were treated by common dose of methylene blue and Mei Ting respectively(P >0.05),the same as 2 times dose of methylene blue,but it changed when combining with EDTA.And the extracellular maxtrix of Saprolegnia biofilm reduced after being treated by EDTA.In conclusion,EDTA could destroy the structure of biofilm.The combination of EDTA with anti-Saproleg-nia drugs had significant activity to remove Saprolegnia biofilm.
出处
《动物医学进展》
北大核心
2015年第8期55-58,共4页
Progress In Veterinary Medicine
基金
国家863计划项目(2011AA10A216)
上海市科技兴农重点攻关项目(沪农科攻字(2011)第4-8号)
国家水产种质资源平台运行服务项目
上海高校知识服务平台资助项目
关键词
水霉菌
生物膜
乙二胺四乙酸
抗菌药
联合用药
Saprolegnia
biofilm
ethylenediaminetetraacetic acid
antimicrobial agent
drug combination
