Ding Chengshi, Pan Jie, Jin Min, Yang Dong, Shen Zhiqiang, Wang Jingfeng, Zhang Bin, Liu Weili, Fu Jialun, Guo Xuan, Wang Daning, Chen Zhaoli, Yin Jing, Qiu Zhigang, Li Junwen
a College of Life Science, Shandong Normal University , Jinan , China ;
b College of Life Science, Zaozhuang University , Zaozhuang , China ;
Nanotoxicology. 2016 Oct;10(8):1051-60. doi: 10.3109/17435390.2016.1161856. Epub 2016 Apr 6.
Nanomaterial pollution and the spread of antibiotic resistance genes (ARGs) are global public health and environmental concerns. Whether nanomaterials could aid the transfer of ARGs released from dead bacteria into live bacteria to cause spread of ARGs is still unknown. Here, we demonstrated that nano-Al2O3 could significantly promote plasmid-mediated ARGs transformation into Gram-negative Escherichia coli strains and into Gram-positive Staphylococcus aureus; however, bulk Al2O3 did not have this effect. Under suitable conditions, 7.4 × 10(6) transformants of E. coli and 2.9 × 10(5) transformants of S. aureus were obtained from 100 ng of a pBR322-based plasmid when bacteria were treated with nano-Al2O3. Nanoparticles concentrations, plasmid concentrations, bacterial concentrations, interaction time between the nanomaterial and bacterial cells and the vortexing time affected the transformation efficiency. We also explored the mechanisms underlying this phenomenon. Using fluorescence in situ hybridization and scanning electron microscopy, we found that nano-Al2O3 damaged the cell membrane to produce pores, through which plasmid could enter bacterial cells. Results from reactive oxygen species (ROS) assays, genome-wide expression microarray profiling and quantitative real-time polymerase chain reactions suggested that intracellular ROS damaged the cell membrane, and that an SOS response promoted plasmid transformation. Our results indicated the environmental and health risk resulting from nanomaterials helping sensitive bacteria to obtain antibiotic resistance.
纳米材料污染和抗生素抗性基因(ARGs)的传播是全球公共卫生和环境领域关注的问题。纳米材料是否会促进死亡细菌释放的ARGs转移到活细菌中从而导致ARGs传播仍不清楚。在此,我们证明了纳米Al2O3能够显著促进质粒介导的ARGs转化到革兰氏阴性大肠杆菌菌株和革兰氏阳性金黄色葡萄球菌中;然而,块状Al2O3没有这种作用。在合适的条件下,当用纳米Al2O3处理细菌时,从100 ng基于pBR322的质粒中可获得7.4×10(6)个大肠杆菌转化子和2.9×10(5)个金黄色葡萄球菌转化子。纳米颗粒浓度、质粒浓度、细菌浓度、纳米材料与细菌细胞之间的相互作用时间以及涡旋时间都会影响转化效率。我们还探究了这一现象背后的机制。通过荧光原位杂交和扫描电子显微镜,我们发现纳米Al2O3破坏细胞膜形成孔洞,质粒可通过这些孔洞进入细菌细胞。活性氧(ROS)检测、全基因组表达微阵列分析和定量实时聚合酶链反应的结果表明,细胞内ROS破坏细胞膜,并且SOS反应促进了质粒转化。我们的结果表明纳米材料帮助敏感细菌获得抗生素抗性会带来环境和健康风险。