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Selective pressure of antibiotics on ARGs and bacterial communities in manure-polluted freshwater-sediment microcosms

机译:粪便污染的淡水沉积物缩微中抗生素对ARG和细菌群落的选择压力

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摘要

The aim of this study was to investigate selective pressure of antibiotics on antibiotic resistance genes (ARGs) and bacterial communities in manure-polluted aquatic environment. Three treatment groups were set up in freshwater-sediment microcosms: tetracyclines group, sulfonamides group and fluoroquinolones group. Sediment and water samples were collected on day 14 after treatment. Antibiotic concentrations, ARGs abundances and bacterial community composition were analyzed. Antibiotic concentrations were determined by ultra-performance liquid chromatography-electrospray tandem mass spectrometry. ARGs abundances were quantified by real time quantitative PCR. Bacterial community composition was analyzed based on amplicon sequencing. Of the three classes of antibiotics analyzed in the treatment groups, accumulation amounts were tetracyclines> fluoroquinolone> sulfonamides in the sediment samples, while they were sulfonamides> fluoroquinolone> tetracyclines in the water samples. In the treatment groups, the relative abundances of some tet resistance genes [tet(W) and tet(X)] and plasmid-mediated quinolone resistance (PMQR) genes [oqx(B) and aac(6′)-Ib] in sediment samples were significantly higher than those in the paired water samples. Tetracyclines significantly selected the bacterial classes including Gammaproteobacteria, Clostridia, and the genera including Salmonella, Escherichia/Shigella, Clostridium, Stenotrophomonas in sediment samples. The significant selection on bacterial communities posed by sulfonamides and fluoroquinolones was also observed. The results indicated that sediment may supply an ideal setting for maintenance and persistence of tet resistance genes [tet(W) and tet(X)] and PMQR genes [oqx(B) and aac(6′)-Ib] under antibiotic pollution. The results also highlighted that antibiotics significantly selected specific bacterial communities including the taxa associated with opportunistic pathogens.
机译:这项研究的目的是调查在粪便污染的水生环境中抗生素对抗生素抗性基因(ARGs)和细菌群落的选择性压力。淡水沉积物缩微分为三个治疗组:四环素类,磺酰胺类和氟喹诺酮类。处理后第14天收集沉积物和水样。分析了抗生素浓度,ARGs丰度和细菌群落组成。通过超高效液相色谱-电喷雾串联质谱法测定抗生素浓度。通过实时定量PCR定量ARG的丰度。基于扩增子测序分析细菌群落组成。在治疗组中分析的三类抗生素中,沉积物样品中的累积量为四环素>氟喹诺酮>磺酰胺,而水样品中的累积量为磺酰胺>氟喹诺酮>四环素。在治疗组中,沉积物中某些tet抗性基因[tet(W)和tet(X)]和质粒介导的喹诺酮抗性(PMQR)基因[oqx(B)和aac(6')-Ib]的相对丰度样品明显高于配对的水样品中的样品。四环素显着选择细菌种类,包括γ-变形杆菌,梭状芽胞杆菌,以及沉积物样品中的沙门氏菌,大肠埃希氏菌/志贺氏菌,梭菌,嗜单胞菌属。还观察到磺酰胺和氟喹诺酮类药物对细菌群落的显着选择。结果表明,沉积物可能为抗生素污染下tet抗性基因[tet(W)和tet(X)]和PMQR基因[oqx(B)和aac(6')-Ib]的维持和持久性提供理想的环境。结果还突出显示,抗生素显着选择了特定的细菌群落,包括与机会病原体相关的分类单元。

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