Assessment of Doñana National Park contamination in Procambarus clarkii: Integration of conventional biomarkers and proteomic approaches |
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Authors: | Amalia Vioque-Fernández Juan López-Barea |
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Affiliation: | a Department of Biochemistry and Molecular Biology, Severo Ochoa building, Campus of Rabanales, University of Córdoba, A4 highway, Km 396a, 14071 Córdoba, Spain b Department of Chemistry and Environmental Sciences, IBILCE-UNESP, Cristóvão Colombo street 2265, 15054-000, São José do Rio Preto, São Paulo, Brazil |
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Abstract: | 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. |
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Keywords: | AChE acetylcholinesterase AJO &ldquo Ajolí &rdquo BER &ldquo Bernabe&rdquo CAT catalase CbE carboxylesterase CDNB 1-chloro-2 4-dinitrobenzene 2-DE two-dimensional gel electrophoresis DNP Doñ ana National Park DTNB 5 5-dithiobis-2-dinitrobenzoic acid DTT dithiothreitol EDTA ethylenediaminetetraacetic acid EROD ethoxyresorufine-O-deethylase GLYOX II glyoxalase II GPx glutathione peroxidase G6PDH glucose-6-phosphate dehydrogenase GSH/GSSG reduced/oxidized glutathione GST glutathione S-transferase IEF isoelectrofocusing LD laguna Dulce&rdquo LDP &ldquo lucio del Palacio&rdquo MAT &ldquo el Matochal&rdquo MDA malondialdehyde MT metallothionein NADPH β-nicotinamideadenine dinucleotide phosphate reduced PAR &ldquo Partido&rdquo PEPs protein expression profiles PMSF phenylmethanesulfonylfluoride ROC &ldquo Rocina&rdquo SDS sodium dodecyl sulphate SDS-PAGE polyacrylamide gel electrophoresis with SDS VIL &ldquo Villafranco&rdquo |
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