CIIMAR - Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Laboratory of Ecotoxicology and Ecology, Rua dos Bragas 289, 4050-123 Porto, Portugal.
Sci Total Environ. 2013 Jan 15;443:454-63. doi: 10.1016/j.scitotenv.2012.10.077. Epub 2012 Dec 3.
Fluoranthene (FLU) is a priority polycyclic aromatic hydrocarbon (PAH) commonly detected in estuarine sediments, water and biota. Despite this, information on FLU detection, accumulation and effects on marine crustaceans is scarce. This work investigated the accumulation of FLU in Carcinus maenas and the responses of several early-warning biomarkers after a 7-day laboratory exposure to five FLU concentrations (2.56 to 100 μg L(-1)). After exposure to FLU, sub-samples of the crabs' digestive gland and muscle were collected for biomarker determinations. The remaining digestive gland and muscle, together with the rest of the whole-body soft tissues, were analysed for FLU residues by gas chromatography-mass spectrometry (GC-MS). The biomarkers assessed were: i) the quantification of FLU-type compounds by fixed wavelength fluorescence (FF); ii) the activities of glutathione S-transferases (GST) and glutathione reductase (GR), and the levels of total glutathione (GT) and lipid peroxidation (LPO) for oxidative stress; iii) the activity of acetylcholinesterase (AChE) for neurotoxicity; iv) the activities of isocitrate dehydrogenase (IDH) and lactate dehydrogenase (LDH) enzymes, and total protein, glycogen and lipids as indicators of changes in energy metabolism and storage; and v) the lysosomal membrane stability (LMS) as a measure of cell damage. The results showed strong (R(2)>0.95) concentration-dependent accumulation of FLU residues (as measured by GC-MS) in the remaining whole-body soft tissues and of FLU-type compounds (as measured by FF) in the digestive gland and muscle. A strong positive linear relationship (R(2)=0.91) between FLU residues and FLU-type compounds was also found. Comparing to controls, activities of GST and GR were significantly higher in crabs exposed to ≥16 and ≥40 μg L(-1) FLU, respectively. TG levels and IDH activity showed a significant trend to increase with FLU concentrations whereas AChE activity exhibited the opposite trend. FF measurements in the digestive gland and muscle proved to be an expeditious cost-effective method to assess the uptake and availability of FLU and its metabolites in C. maenas. The results suggest that under continuous environmental exposure, FLU may enhance detoxification and anti-oxidant defences, and cause alterations in the aerobic energy pathway, as well as neuromuscular toxic effects that may increase C. maenas risk of predation.
荧蒽(FLU)是一种优先多环芳烃(PAH),通常在河口沉积物、水和生物群中检测到。尽管如此,关于荧蒽的检测、积累以及对海洋甲壳类动物的影响的信息仍然很少。本研究调查了荧蒽在螃蟹(Carcinus maenas)中的积累情况,以及在 7 天实验室暴露于 5 种荧蒽浓度(2.56 至 100 μg L(-1))后,几种早期预警生物标志物的反应。暴露于荧蒽后,采集螃蟹的消化腺和肌肉的亚样本进行生物标志物测定。其余的消化腺和肌肉,以及整个身体软组织的其余部分,通过气相色谱-质谱法(GC-MS)分析荧蒽残留。评估的生物标志物为:i)通过固定波长荧光(FF)定量荧蒽型化合物;ii)谷胱甘肽 S-转移酶(GST)和谷胱甘肽还原酶(GR)的活性,以及总谷胱甘肽(GT)和脂质过氧化(LPO)的水平以评估氧化应激;iii)乙酰胆碱酯酶(AChE)的活性以评估神经毒性;iv)异柠檬酸脱氢酶(IDH)和乳酸脱氢酶(LDH)的活性以及总蛋白、糖原和脂质作为能量代谢和储存变化的指标;以及 v)溶酶体膜稳定性(LMS)作为细胞损伤的衡量标准。结果表明,GC-MS 测量的剩余全身软组织中荧蒽残留(荧蒽型化合物)以及 FF 测量的消化腺和肌肉中荧蒽型化合物(荧蒽型化合物)均表现出强烈的浓度依赖性积累(R(2)>0.95)。还发现荧蒽残留与荧蒽型化合物之间存在强烈的正线性关系(R(2)=0.91)。与对照组相比,暴露于≥16 和≥40 μg L(-1)荧蒽的螃蟹中 GST 和 GR 的活性分别显著升高。TG 水平和 IDH 活性呈明显的随荧蒽浓度增加的趋势,而 AChE 活性则呈相反的趋势。荧光法测量消化腺和肌肉,被证明是一种快捷、经济有效的方法,可用于评估 C. maenas 中荧蒽及其代谢物的摄取和可用性。结果表明,在持续的环境暴露下,荧蒽可能会增强解毒和抗氧化防御能力,并导致有氧能量途径的改变,以及肌肉神经毒性效应,这可能会增加 C. maenas 被捕食的风险。