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Fate of Extracellular DNA in the Production of Fertilizers from Source-Separated Urine.
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2020-01-22 , DOI: 10.1021/acs.est.9b04263 Heather E Goetsch 1 , Nancy G Love 1 , Krista R Wigginton 1
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2020-01-22 , DOI: 10.1021/acs.est.9b04263 Heather E Goetsch 1 , Nancy G Love 1 , Krista R Wigginton 1
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The practice of urine source-separation for fertilizer production necessitates an understanding of the presence and impact of extracellular DNA in the urine. This study examines the fate of plasmid DNA carrying ampicillin and tetracycline resistance genes in aged urine, including its ability to be taken up and expressed by competent bacteria. Plasmid DNA incubated in aged urine resulted in a >2 log loss of bacterial transformation efficiency in Acinetobacter baylyi within 24 h. The concentration of ampicillin and tetracycline resistance genes, as measured with quantitative polymerase chain reaction, did not correspond with the observed transformation loss. When the plasmid DNA was incubated in aged urine that had been filtered (0.22 μm) or heated (75 °C), the transformation efficiencies were more stable than when the plasmids were incubated in unfiltered and unheated aged urine. Gel electrophoresis results indicated that plasmid linearization by materials larger than 100 kDa in the aged urine caused the observed transformation efficiency decreases. The results of this study suggest that extracellular DNA released into aged urine poses a low potential for the spread of antibiotic resistance genes to bacteria once it is released to the environment.
中文翻译：
从源分离尿液生产肥料时细胞外DNA的命运。
尿液源分离生产肥料的实践需要了解尿液中细胞外DNA的存在及其影响。这项研究检查了在老年尿液中携带氨苄青霉素和四环素抗性基因的质粒DNA的命运，包括其被感受态细菌吸收和表达的能力。在老化的尿液中孵育的质粒DNA在24小时内导致Baylyi不动杆菌中细菌转化效率的损失> 2 log。用定量聚合酶链反应测量的氨苄青霉素和四环素抗性基因的浓度与观察到的转化损失不对应。当质粒DNA在已过滤（0.22μm）或加热（75°C）的老化尿液中孵育时，与将质粒在未过滤和未加热的老化尿液中孵育相比，转化效率更加稳定。凝胶电泳结果表明，老化尿液中大于100 kDa的物质使质粒线性化，导致观察到的转化效率降低。这项研究的结果表明，释放到老年尿液中的细胞外DNA一旦释放到环境中，对细菌产生抗药性基因的可能性就很小。
更新日期：2020-01-23
中文翻译：
从源分离尿液生产肥料时细胞外DNA的命运。
尿液源分离生产肥料的实践需要了解尿液中细胞外DNA的存在及其影响。这项研究检查了在老年尿液中携带氨苄青霉素和四环素抗性基因的质粒DNA的命运，包括其被感受态细菌吸收和表达的能力。在老化的尿液中孵育的质粒DNA在24小时内导致Baylyi不动杆菌中细菌转化效率的损失> 2 log。用定量聚合酶链反应测量的氨苄青霉素和四环素抗性基因的浓度与观察到的转化损失不对应。当质粒DNA在已过滤（0.22μm）或加热（75°C）的老化尿液中孵育时，与将质粒在未过滤和未加热的老化尿液中孵育相比，转化效率更加稳定。凝胶电泳结果表明，老化尿液中大于100 kDa的物质使质粒线性化，导致观察到的转化效率降低。这项研究的结果表明，释放到老年尿液中的细胞外DNA一旦释放到环境中，对细菌产生抗药性基因的可能性就很小。
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