Laboratório de Biologia Molecular, NUPEM/UFRJ, Universidade Federal do Rio de Janeiro, Macae, Brazil,
Ecotoxicology. 2013 May;22(4):642-55. doi: 10.1007/s10646-013-1055-z. Epub 2013 Mar 13.
Amphipod crustaceans have been widely used as invertebrate models in ecotoxicology due to their importance in the food chain. However, few studies have evaluated the genotoxic effects of pollutants in this model using the comet assay. The main obstacle to using amphipods in the comet assay is the difficulty in obtaining enough blood cells from a single individual. In this study, we evaluated the genotoxic effects of the water-soluble fraction (WSF) of heavy oil on the brackish/freshwater amphipod Quadrivisio aff. lutzi, which is common in the coastal lagoons of southeastern Brazil, using hemocytes obtained from single amphipods (without pooling) after optimizing hemolymph extraction. The comet assay revealed significantly higher DNA damage levels (2- to 6-fold higher) in treated amphipods compared to untreated ones with a sublethal concentration of 17.6 % of the WSF within 72 h of treatment. Two independent experiments confirmed an "up and down" pattern of DNA damage, measured as the % of DNA contained in the tail of the comets. Elevations in DNA damage levels were observed at the 6 and 48 h time points, while very low levels of DNA damage were observed at the 24 and 72 h time points. Furthermore, the comet assay revealed gender variability in the levels of DNA damage after short-term exposure.
由于在食物链中的重要地位,端足目甲壳动物已被广泛用作生态毒理学中的无脊椎动物模型。然而,使用彗星试验评估这种模型中污染物的遗传毒性影响的研究较少。在彗星试验中使用端足目动物的主要障碍是从单个个体中获得足够血细胞的困难。在这项研究中,我们评估了水溶性馏分(WSF)对在巴西东南部沿海泻湖中常见的半咸水/淡水端足目甲壳动物 Quadrivisio aff. lutzi 的遗传毒性影响,使用优化的血淋巴提取方法从单个端足目动物(不混合)中获得的血细胞进行实验,而无需混合。彗星试验显示,在处理的端足目动物中,与未经处理的端足目动物相比,具有亚致死浓度(WSF 的 17.6%)的处理组中 DNA 损伤水平显著更高(高 2-6 倍),在处理后 72 小时内。两项独立的实验证实了 DNA 损伤的“上升和下降”模式,其表现为彗星尾部包含的 DNA 百分比。在 6 和 48 小时时间点观察到 DNA 损伤水平的升高,而在 24 和 72 小时时间点观察到非常低水平的 DNA 损伤。此外,彗星试验显示在短期暴露后,性别差异会影响 DNA 损伤水平。