Detection of antibiotic resistance genes in wastewater treatment plant – molecular and classical approach
Detection of antibiotic resistance genes in wastewater treatment plant – molecular and classical approach
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废水处理厂中抗生素抗性基因的检测——分子和经典方法
DOI：
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
J. Surmacz
中科院分区：
文献类型：
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作者：
A. Ziembińska;E. Felis;J. Folkert;Anna Meresta;D. Stawicka;A. Gnida;J. Surmacz
摘要抗生素是一类对环境有潜在危害的物质。它们可以在致病性和非致病性细菌之间的细菌耐药性转移中发挥作用。在该实验中，使用了三种代表性的医学上重要的化学治疗剂，其经HPLC分析证实以高浓度存在于废水处理厂中：红霉素、磺胺甲恶唑和甲氧苄啶。活性污泥中的红霉素浓度不高于20 ng L−1。N-乙酰磺胺甲恶唑浓度在冬季为3349 ± 719 ng L − 1，夏季为2933 ± 429 ng L−1。甲氧苄啶在废水中的浓度在冬季和夏季分别为400 ± 22和364 ± 60 ng L−1。由于对这些物质的PCR可检测耐药机制多种多样，因此选择了文献中最常见的耐药机制。对于红霉素：在甲氧苄啶耐药的情况下，在本研究中使用了磺胺甲恶唑：sul 1、sul 2、sul 3基因的dhfrA 1和dhfr 14。在从活性污泥中分离的纯菌株和活性污泥样品本身中分析抗性基因的存在。研究表明，最小抑菌浓度（MIC）值与预期存在的一种以上的耐药机制不一致。大多数分离株只具有一种负责特定化疗耐药性的基因。结果表明，利用PCR技术可以直接检测活性污泥中抗性基因的存在。由于实验中使用的分离株数量有限，这些结果应被视为初步结果。
Abstract Antibiotics are a group of substances potentially harmful to the environment. They can play a role in bacterial resistance transfer among pathogenic and non-pathogenic bacteria. In this experiment three representatives of medically important chemotherapeutics, confirmed to be present in high concentrations in wastewater treatment plants with HPLC analysis were used: erythromycin, sulfamethoxazole and trimethoprim. Erythromycin concentration in activated sludge was not higher than 20 ng L−1. N-acetylo-sulfamethoxazole concentration was 3349 ± 719 in winter and 2933 ± 429 ng L−1 in summer. Trimethoprim was present in wastewater at concentrations 400 ± 22 and 364 ± 60 ng L−1, respectively in winter and summer. Due to a wide variety of PCR-detectable resistance mechanisms towards these substances, the most common found in literature was chosen. For erythromycin: erm and mef genes, for sulfamethoxazole: sul1, sul2, sul3 genes, in the case of trimethoprim resistance dhfrA1 and dhfr14 were used in this study. The presence of resistance genes were analyzed in pure strains isolated from activated sludge and in the activated sludge sample itself. The research revealed that the value of minimal inhibitory concentration (MIC) did not correspond with the expected presence of more than one resistance mechanisms. Most of the isolates possessed only one of the genes responsible for a particular chemotherapeutic resistance. It was confirmed that it is possible to monitor the presence of resistance genes directly in activated sludge using PCR. Due to the limited isolates number used in the experiment these results should be regarded as preliminary.