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Recombinant luminescent bacterial sensors for the measurement of bioavailability of cadmium and lead in soils polluted by metal smelters

机译:重组发光细菌传感器,用于测量金属冶炼厂污染的土壤中镉和铅的生物利用度

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摘要

Environmental hazard of heavy metals in soils depends to a large extent on their bioavailability. The approach used in this study enables the determination of bioavailable metals in solid-phase samples. Two recombinant bacterial sensors, one responding specifically to cadmium and the other to lead and cadmium by increase of luminescence (firefly luciferase was used as a reporter) were used to determine the bioavailability of these metals in soil-water suspensions (a contact assay) and respective particle-free extracts. Fifty agricultural soils sampled near zinc and lead smelters in the Northern France containing up to (mg/kg) 20.1 of Cd, 1050 of Pb and 1390 of Zn were analysed. As the soil matrix interferes with the assay, recombinant luminescent control bacteria lacking the metal recognizing protein and corresponding promoter (thus, being not metal-inducible) but otherwise comparable to the sensor bacteria (the same host bacterium and plasmid encoding luciferase) were used in parallel to take into account the possible quenching and/or stimulating effects of the sample on the luminescence of the sensor bacteria. Both, chemical and sensor analysis showed that only μg/l levels of metals were extracted from the soil into the water phase (0.1% of the total Cd, 0.07% of Pb and 0.5% of Zn). However, 115-fold more Cd and 40-fold more Pb proved bioavailable if the sensor bacteria were incubated in soil suspensions (i.e., in the contact assay). The bioavailability of metals in different soils varied (depending probably on soil type) ranging from 0.5% to 56% for cadmium and from 0.2% to 8.6% for lead.
机译:土壤中重金属的环境危害在很大程度上取决于其生物利用度。本研究中使用的方法能够测定固相样品中的生物利用金属。使用两个重组细菌传感器,一个专门对镉作出反应,另一个通过发光增加对铅和镉作出反应(萤火虫荧光素酶用作报告基因)用于测定这些金属在土壤-水悬浮液中的生物利用度(接触测定)和各自的无颗粒提取物。在法国北部的锌和铅冶炼厂附近对五十种农业土壤进行了采样分析,这些土壤中的镉含量高达(mg / kg)20.1,镉含量为1050,铅含量为1390。由于土壤基质会干扰测定,因此重组荧光控制细菌缺乏金属识别蛋白和相应的启动子(因此不是金属诱导型的),但在其他方面与传感器细菌(相同的宿主细菌和编码荧光素酶的质粒)相当。同时考虑到样品对传感器细菌发光的可能的淬灭和/或刺激作用。化学和传感器分析均显示,只有微克/升的金属从土壤中提取到水相中(总Cd的0.1%,Pb的0.07%和Zn的0.5%)。但是,如果将传感器细菌在土壤悬浮液中孵育(即在接触分析中),则可证明Cd具有115倍的Cd和40倍于Pb的生物利用率。金属在不同土壤中的生物利用度变化(可能取决于土壤类型)对于镉而言为0.5%至56%,对于铅而言为0.2%至8.6%。

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