Biomonitoring requires the application of batteries of different biomarkers, as environmental contaminants induce multiple responses in organisms that are not necessarily correlated. Omic technologies were proposed as an alternative to conventional biomarkers since these techniques quantitatively monitor many biological molecules in a high-throughput manner and thus provide a general appraisal of biological responses altered by exposure to contaminants. As the studies using omic technologies increase, it is becoming clear that any single omic approach may not be sufficient to characterize the complexity of ecosystems. This work aims to provide a preliminary working scheme for the use of combined transcriptomic and proteomic methodologies in environmental biomonitoring. There are difficulties in working with nonmodel organisms as bioindicators when combining several omic approaches. As a whole, our results with heterologous microarrays in M. spretus and suppressive subtractive hybridization (SSH) in P. clarkii indicated that animals sustaining a heavy pollution burden exhibited an enhanced immune response and/or cell apoptosis. The proteomic studies, although preliminary, provide a holistic insight regarding the manner by which pollution shifts protein intensity in two-dimensional gel electrophoresis (2-DE), completing the transcriptomic approach. In our study, the sediment element concentration was in agreement with the intensity of protein expression changes in C. maenas crabs. In conclusion, omics are useful technologies in addressing environmental issues and the determination of contamination threats.
In vivo effects of two sublethal doses of chlorpyrifos and carbaryl were studied in Procambarus clarkii after 2 and 7 days of exposure, and after pesticide removal. Chlorpyrifos inhibited carboxylesterase activity in a concentration-dependent manner, but acetylcholinesterase was less sensitive. Compared with chlorpyrifos, carbaryl had a less marked effect on esterase activity. The effects of selected pesticides on biotransformation or oxidative stress biomarkers were contradictory. Chlorpyrifos lowered ethoxyresorufin-O-deethylase (EROD), catalase and oxidized glutathione (GSSG) levels but raised glutathione-S-transferase activity, while carbaryl raised EROD, catalase and glutathione-S-transferase, but lowered glutathione peroxidase and reduced glutathione (GSH) levels. The effects on protein expression patterns depending on pesticide type and the tissue used for analysis were studied in parallel by 2-DE. In gill and nervous tissue about 2000 spots (pI 4-7) were resolved, with quite different expression patterns. Chlorpyrifos altered 72 proteins, mostly in nervous tissue, and carbaryl 35, distributed evenly between organs. Several specific spots were selected as specific protein expression signatures for chlorpyrifos or carbaryl exposure in gills and nervous tissue, respectively.
A speciation approach based on orthogonal chromatographic systems coupled to inductively coupled plasma mass spectrometry (ICP-MS) was used to characterise the biological response of free-living mice Mus spretus to environmental pollution caused by arsenic in different areas of the Doñana National Park (south-west Spain). The relative presence of inorganic and organic forms of arsenic was studied in cytosolic extracts from high metabolic activity organs of Mus spretus mice: kidneys, liver, and brain. An instrumental coupling of size-exclusion chromatography with UV and collision/reaction cell-ICP-MS detectors (SEC-UV-ICP-ORC-MS) both in analytical and preparative scale was used for this purpose. The results showed the presence of low molecular mass (LMM) molecules linked to arsenic in these tissues especially in the kidneys, where the presence of these arsenic metabolites was higher. On the other hand, the presence of these arsenicals varied from one area to the other, which can be related to a different occurrence of contaminants. These low molecular mass fractions were collected by preparative SEC chromatography for later study with ion exchange chromatography and detection by ICP-ORC-MS, using both anionic and cationic columns. The results showed the higher presence of MMA and DMA in kidneys of mice caught in contaminated areas and the existence of small amounts of unidentified arsenicals when cation-exchange chromatography was used, which could be related to the presence of dimethylarsinoylethanol (DMAE), thioarsenic species, or arsenocholine (AsC).
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