畜禽养殖场附近污水中多粘菌素E耐药性肠杆菌的分离分析及内源质粒检测

发布时间：2018-03-01 12:43

本文关键词： 畜禽养殖场肠杆菌多粘菌素E 多重耐药性质粒　出处：《生态环境学报》2017年04期 　论文类型：期刊论文

【摘要】：畜禽养殖业大量使用抗生素引起了广泛的微生物耐药性问题,研究抗生素耐药菌株可以了解环境中细菌的耐药性情况和抗性基因的传播机制。采集畜禽养殖场周边的污水,分析了污水中多粘菌素E耐药性细菌和耐药性肠杆菌的丰度;同时以一株具有较高多粘菌素E耐药水平的肠杆菌R11为例,首先进行16S r DNA测序和抗生素耐药谱分析,然后通过质粒提取、酶切电泳以及脉冲场凝胶电泳(PFGE),分析其内源性质粒等情况。结果表明:在畜禽养殖场周边污水环境中,多粘菌素E耐药性细菌占细菌总数的0.14%~0.35%,多粘菌素E耐药性肠杆菌占肠杆菌总数的1.50%~7.40%,而肠杆菌占细菌总数的1.13%~2.45%,说明肠杆菌可能是携带多粘菌素E抗性基因的主要细菌种类,同时也是养殖业污水中细菌的主要组成部分。16S r DNA序列分析确定R11是一株Enterobacter cloacae,抗生素耐药谱分析R11具有多重耐药性,除对林可霉素敏感外,阿米卡星、四环素和环丙沙星的最小抑菌浓度(MICs)为10μg?m L~(-1),阿莫西林的MICs为25μg?m L~(-1),磺胺甲恶唑的MICs为32μg?m L~(-1),氨苄西林的MICs为40μg?m L~(-1),链霉素的MICs为90μg?m L~(-1),而多粘菌素E的MICs最高,达到96μg?m L~(-1)。质粒提取酶切和PFGE分离鉴定结果显示,R11菌株可能有多个内源性质粒。综上所述,R11菌株具有较高水平的多粘菌素E耐药性,并且具有多重耐药性和多个内源性质粒,对于研究多粘菌素E耐药性机制具有一定的参考价值。
[Abstract]:The extensive use of antibiotics in livestock and poultry breeding has caused a wide range of microbial drug resistance problems. The study of antibiotic resistant strains can understand the drug resistance of bacteria in the environment and the transmission mechanism of resistance genes, and collect sewage from livestock and poultry farms. The abundance of polymyxin E resistant bacteria and resistant Enterobacter spp in sewage was analyzed, and the 16s r DNA sequencing and antibiotic resistance spectrum analysis were carried out with a high polymyxin E resistant Enterobacter coli R11 as an example. Then the plasmid extraction, enzyme digestion electrophoresis and pulse field gel electrophoresis were used to analyze the endogenous plasmids. The results showed that: in the sewage environment around the livestock and poultry farms, Polymyxin E resistant bacteria accounted for 0.14% of the total number of bacteria, polymyxin E resistant Enterobacter accounted for 1.50% of the total number of Enterobacter, and Enterobacter accounted for 1.133.45% of the total number of bacteria, indicating that Enterobacter may be the main type of bacteria carrying polymyxin E resistance gene. At the same time, it is also the main component of bacteria. 16s r DNA sequence analysis confirmed that R11 was a strain of Enterobacter cloacae.The antibiotic resistance spectrum analysis showed that R11 was multidrug resistant, except for lincomycin sensitive, amikacin. The minimum inhibitory concentrations of tetracycline and ciprofloxacin were 10 渭 g? The MICs of amoxicillin is 25 渭 g? The MICs of sulfamethoxazole is 32 渭 g? The MICs of ampicillin is 40 渭 g? The MICs of streptomycin is 90 渭 g? The MICs of polymyxin E was the highest, reaching 96 渭 g? The results of restriction endonuclease digestion and PFGE isolation showed that the strain R11 might have multiple endogenous plasmids. In conclusion, the strain R11 had high level of polymyxin E resistance, and had multiple multidrug resistance and multiple endogenous plasmids. It has certain reference value for studying the mechanism of polymyxin E resistance.
【作者单位】：山东建筑大学市政与环境工程学院;山东省医学科学院山东省医药生物技术研究中心;
【基金】：国家自然科学基金项目(31500094) 山东省自然科学基金项目(ZR2015EM018) 山东省医学科学院医药卫生科技创新工程项目(201604)
【分类号】：X172;X713

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