Huang Kailong, Tang Junying, Zhang Xu-Xiang, Xu Ke, Ren Hongqiang
State Key Laboratory of Pollution Control and Resource Reuse, Environmental Health Research Center, School of the Environment, Nanjing University, Nanjing 210023, China.
Int J Mol Sci. 2014 Jun 5;15(6):10083-100. doi: 10.3390/ijms150610083.
In order to comprehensively investigate tetracycline resistance in activated sludge of sewage treatment plants, 454 pyrosequencing and Illumina high-throughput sequencing were used to detect potential tetracycline resistant bacteria (TRB) and antibiotic resistance genes (ARGs) in sludge cultured with different concentrations of tetracycline. Pyrosequencing of 16S rRNA gene revealed that tetracycline treatment greatly affected the bacterial community structure of the sludge. Nine genera consisting of Sulfuritalea, Armatimonas, Prosthecobacter, Hyphomicrobium, Azonexus, Longilinea, Paracoccus, Novosphingobium and Rhodobacter were identified as potential TRB in the sludge. Results of qPCR, molecular cloning and metagenomic analysis consistently indicated that tetracycline treatment could increase both the abundance and diversity of the tet genes, but decreased the occurrence and diversity of non-tetracycline ARG, especially sulfonamide resistance gene sul2. Cluster analysis showed that tetracycline treatment at subinhibitory concentrations (5 mg/L) was found to pose greater effects on the bacterial community composition, which may be responsible for the variations of the ARGs abundance. This study indicated that joint use of 454 pyrosequencing and Illumina high-throughput sequencing can be effectively used to explore ARB and ARGs in the environment, and future studies should include an in-depth investigation of the relationship between microbial community, ARGs and antibiotics in sewage treatment plant (STP) sludge.
为了全面研究污水处理厂活性污泥中的四环素抗性，采用454焦磷酸测序和Illumina高通量测序技术，检测不同浓度四环素培养的污泥中潜在的四环素抗性细菌（TRB）和抗生素抗性基因（ARG）。16S rRNA基因的焦磷酸测序结果表明，四环素处理对污泥的细菌群落结构有很大影响。在污泥中鉴定出9个属，包括硫杆菌属、武装单胞菌属、柄杆菌属、生丝微菌属、氮单胞菌属、长线菌属、副球菌属、新鞘氨醇菌属和红杆菌属，它们被认为是潜在的TRB。qPCR、分子克隆和宏基因组分析结果一致表明，四环素处理可增加tet基因的丰度和多样性，但会降低非四环素ARG的出现频率和多样性，尤其是磺胺抗性基因sul2。聚类分析表明，亚抑制浓度（5mg/L）的四环素处理对细菌群落组成的影响更大，这可能是ARG丰度变化的原因。本研究表明，联合使用454焦磷酸测序和Illumina高通量测序可有效用于探索环境中的ARB和ARG，未来的研究应深入调查污水处理厂（STP）污泥中微生物群落、ARG和抗生素之间的关系。
