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评估细菌生物传感器以确定铬酸盐生物有效性并评估土壤的生态毒性。

Evaluation of bacterial biosensors to determine chromate bioavailability and to assess ecotoxicity of soils.

作者信息

Coelho Catarina, Branco Rita, Natal-da-Luz Tiago, Sousa José Paulo, Morais Paula V

机构信息

IMAR-CMA, 3004-517 Coimbra, Portugal.

IMAR-CMA, 3004-517 Coimbra, Portugal; Department of Life Sciences, FCTUC, University of Coimbra, 3001-401 Coimbra, Portugal.

出版信息

Chemosphere. 2015 Jun;128:62-9. doi: 10.1016/j.chemosphere.2014.12.026. Epub 2015 Feb 2.

Abstract

Chromate can be considered a potent environmental contaminant and consequently, an understanding of chromate availability and toxicity to soil biology is essential for effective ecological assessment of metal impact in soils. This study shows the response of two bacterial bioreporters, pCHRGFP1 Escherichiacoli and pCHRGFP2 Ochrobactrumtritici, to increasing concentrations of chromate in two different soils. The bioreporters, carrying the regulatory gene chrB transcriptionally fused to the gfp reporter system, exhibited different features. In both, the fluorescence signal and the chromate concentration could be linearly correlated but E. coli biosensor functioned within the range of 0.5-2 μM and O. tritici biosensor within 2-10 μM chromate. The bioreporters were validated through comparative measurements using the chemical chromate methods of diphenylcarbazide and ionic chromatography. The bacterial sensors were used for the estimation of bioavailable fraction of chromate in a natural soil and OECD artificial soil, both spiked with chromate in increasing concentrations of 0-120 mg Cr(VI) kg(-1) of soil. OECD soil showed a faster chromate decrease comparing to the natural soil. The toxicity of soils amended with chromate was also evaluated by ecotoxicological tests through collembolan reproduction tests using Folsomia candida as test organism. Significant correlations were found between collembolans reproduction and chromate concentration in soil (lower at high chromate concentrations) measured by biosensors. Data obtained showed that the biosensors tested are sensitive to chromate presence in soil and may constitute a rapid and efficient method to measure chromate availability in soils.

摘要

铬酸盐可被视为一种强效的环境污染物,因此,了解铬酸盐对土壤生物学的有效性和毒性对于有效评估土壤中金属影响的生态状况至关重要。本研究展示了两种细菌生物报告器,即携带绿色荧光蛋白基因的大肠杆菌pCHRGFP1和小麦苍白杆菌pCHRGFP2,对两种不同土壤中铬酸盐浓度增加的响应。这些携带与绿色荧光蛋白报告系统转录融合的调控基因chrB的生物报告器表现出不同的特性。在这两种生物报告器中,荧光信号与铬酸盐浓度均可呈线性相关,但大肠杆菌生物传感器在0.5 - 2 μM的铬酸盐浓度范围内起作用,而小麦苍白杆菌生物传感器在2 - 10 μM的铬酸盐浓度范围内起作用。通过使用二苯卡巴肼化学铬酸盐法和离子色谱法进行对比测量,对生物报告器进行了验证。将这些细菌传感器用于估计天然土壤和经合组织(OECD)人工土壤中铬酸盐的生物可利用部分,这两种土壤均添加了浓度为0 - 120 mg Cr(VI) kg(-1)土壤且浓度不断增加的铬酸盐。与天然土壤相比,经合组织土壤中铬酸盐的减少速度更快。还通过生态毒理学测试,以白色符氏跳虫为受试生物进行弹尾虫繁殖测试,评估了添加铬酸盐的土壤的毒性。生物传感器测得的弹尾虫繁殖与土壤中铬酸盐浓度(在高铬酸盐浓度下较低)之间存在显著相关性。所获得的数据表明,测试的生物传感器对土壤中铬酸盐的存在敏感,可能构成一种快速有效的方法来测量土壤中铬酸盐的有效性。

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