Vioque-Fernández Amalia, Alves de Almeida Eduardo, López-Barea Juan
Department of Biochemistry and Molecular Biology, Severo Ochoa building, Campus of Rabanales, University of Córdoba, A4 highway, Km 396a, 14071 Córdoba, Spain.
Sci Total Environ. 2009 Feb 15;407(5):1784-97. doi: 10.1016/j.scitotenv.2008.11.051. Epub 2008 Dec 25.
Aquatic ecosystems of Doñana National Park (DNP) were monitored using Procambarus clarkii as bioindicator in four campaigns carried out between 2003 and 2004 to assess environmental quality possibly threatened by agrochemicals used in nearby areas. An integrated approach was carried out, by combining the responses of well-established biomarkers and the massive analysis of biological effects at the proteomic level. In sites potentially polluted, lower catalase, glucose-6-P dehydrogenase, and esterase activities, and higher malondialdehyde, metallothionein and glutathione levels were found. Two-dimensional gel electrophoresis resolved >2500 gill spots, and image analysis detected that 35 showed significant intensity differences between the reference site and the other seven sites studied. The superiority of proteomic approaches was clearly recognized in our study since four different protein expression patterns were established based in the fold-number of up-/down-regulation of the 35 differentially expressed proteins. Sites located within Doñana Biological Reserve were essentially free of contaminants and those near the DNP limits were only slightly polluted. The higher proteomic responses found at the upper "Rocina" and "Partido" courses indicate that non-persistent agrochemicals are mainly used in Doñana surroundings. The highest responses corresponded to rice growing areas placed between the Guadiamar stream and the Guadalquivir River, according to the extended and intensive use of agrochemicals in such areas.
在2003年至2004年期间开展的四项活动中，以克氏原螯虾作为生物指示剂，对多尼亚纳国家公园（DNP）的水生生态系统进行了监测，以评估可能受到附近地区使用的农用化学品威胁的环境质量。通过结合成熟生物标志物的反应和蛋白质组水平上生物效应的大规模分析，采用了一种综合方法。在潜在污染的地点，发现过氧化氢酶、葡萄糖-6-磷酸脱氢酶和酯酶活性较低，而丙二醛、金属硫蛋白和谷胱甘肽水平较高。二维凝胶电泳解析出>2500个鳃部斑点，图像分析检测到35个斑点在参考位点与其他七个研究位点之间存在显著的强度差异。在我们的研究中，蛋白质组学方法的优越性得到了明确认可，因为基于35种差异表达蛋白质的上调/下调倍数建立了四种不同的蛋白质表达模式。位于多尼亚纳生物保护区内的地点基本没有污染物，而靠近DNP边界的地点仅受到轻微污染。在“罗西纳”和“帕尔蒂多”上游河段发现的较高蛋白质组反应表明，多尼亚纳周边地区主要使用非持久性农用化学品。根据此类地区农用化学品的广泛和密集使用情况，最高反应对应于瓜迪亚马尔河和瓜达尔基维尔河之间的水稻种植区。
