IBB-Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Av. Rovisco Pais, 1049-001 Lisboa, Portugal.
IMAR-Instituto do Mar & MARE-Marine and Environmental Sciences Centre, Department of Life Sciences, University of Coimbra, Apartado 3046, 3001-401 Coimbra, Portugal.
Sci Total Environ. 2015 Feb 1;505:161-71. doi: 10.1016/j.scitotenv.2014.09.094. Epub 2014 Oct 14.
The present study is aimed at evaluating whether a gene expression assay with the microbial eukaryotic model Saccharomyces cerevisiae could be used as a suitable warning tool for the rapid preliminary screening of potential toxic effects on organisms due to scenarios of soil and water contamination with pyrimethanil. The assay consisted of measuring changes in the expression of the selected pyrimethanil-responsive genes ARG3 and ARG5,6 in a standardized yeast population. Evaluation was held by assessing the toxicity of surface runoff, a major route of pesticide exposure in aquatic systems due to non-point-source pollution, which was simulated with a pyrimethanil formulation at a semifield scale mimicking worst-case scenarios of soil contamination (e.g. accident or improper disposal). Yeast cells 2-h exposure to the runoff samples led to a significant 2-fold increase in the expression of both indicator genes. These results were compared with those from assays with organisms relevant for the aquatic and soil compartments, namely the nematode Caenorhabditis elegans (reproduction), the freshwater cladoceran Daphnia magna (survival and reproduction), the benthic midge Chironomus riparius (growth), and the soil invertebrates Folsomia candida and Enchytraeus crypticus (survival and reproduction). Under the experimental conditions used to simulate accidental discharges into soil, runoff waters were highly toxic to the standard test organisms, except for C. elegans. Overall, results point out the usefulness of the yeast assay to provide a rapid preview of the toxicity level in preliminary screenings of environmental samples in situations of inadvertent high pesticide contamination. Advantages and limitations of this novel method are discussed.
本研究旨在评估利用微生物真核模式生物酿酒酵母的基因表达分析是否可作为一种合适的预警工具,用于快速初步筛选因嘧啶苯胺污染土壤和水而对生物体产生潜在毒性影响的情况。该分析包括测量在标准化酵母群体中选择的嘧啶苯胺响应基因 ARG3 和 ARG5,6 的表达变化。通过评估半田间规模模拟土壤污染最恶劣情况(例如事故或不当处理)的嘧啶苯胺制剂的地表径流(由于非点源污染,是水生系统中农药暴露的主要途径)的毒性来进行评估。酵母细胞暴露于径流样品 2 小时后,两个指示基因的表达均显著增加了 2 倍。将这些结果与与水生和土壤区室相关的生物体的测定结果进行比较,即线虫秀丽隐杆线虫(繁殖)、淡水枝角类大型蚤(生存和繁殖)、底栖蝇类摇蚊(生长)以及土壤无脊椎动物 Folsomia candida 和 Enchytraeus crypticus(生存和繁殖)。在用于模拟意外排放到土壤中的实验条件下,除秀丽隐杆线虫外,径流对标准测试生物具有高毒性。总体而言,结果表明,该酵母分析可用于在高浓度农药意外污染情况下,对环境样品的毒性水平进行初步筛选时提供快速预览。讨论了这种新方法的优缺点。