Proteomic changes in <Emphasis Type="Italic">Corbicula fluminea</Emphasis> exposed to wastewater from a psychiatric hospital

The increase use of pharmaceutical compounds in veterinary practice and human population results in the ubiquitous presence of these compounds in aquatic ecosystems. Because pharmaceuticals are highly bioactive, there is concern about their toxicological effects in aquatic organisms. Therefore, the aim of this study was to assess the effects of an effluent from a psychiatric hospital (containing a complex mixture of 25 pharmaceutical compounds from eleven therapeutic classes) on the freshwater clam Corbicula fluminea using a proteomic approach. The exposure of C. fluminea to this complex effluent containing anxiolytics, analgesics, lipid regulators, beta blockers, antidepressants, antiepileptics, antihistamines, antihypertensives, antiplatelets and antiarrhythmics induced protein changes after 1 day of exposure in clam gills and digestive gland more evident in the digestive gland. These changes included increase in the abundance of proteins associated with structural (actin and tubulin), cellular functions (calreticulin, proliferating cell nuclear antigen (PCNA), T complex protein 1 (TCP1)) and metabolism (aldehyde dehydrogenase (ALDH), alcohol dehydrogenase, 6 phosphogluconate dehydrogenase). Results from this study indicate that calreticulin, PCNA, ALDH and alcohol dehydrogenase in the digestive gland and T complex protein 1 (TCP1)) and 6 phosphogluconate dehydrogenase in the gills represent useful biomarkers for the ecotoxicological characterization of psychiatric hospital effluents in this species.     

DNA damage as a biomarker of genotoxic contamination in Mytilus galloprovincialis from the south coast of Portugal

DNA damage was evaluated in the haemolymph of Mytilus galloprovincialis from nine sites along the south coast of Portugal using the comet assay. DNA damage was low, in the same range of sites considered to suffer low impact from genotoxic contaminants. Even so, differences between sites, seasons and genders were found. Highest values were in mussels from the main estuaries and the fishery harbour, reflecting higher genotoxin levels, whereas the lowest values can be used as a baseline for future work. Non-contaminant related factors (e.g. temperature and oxygen) were also shown to influence DNA damage. Between seasons, highest values were in summer related not only to the increase of tourism in this region (～10-fold), but also to temperature. Between genders, males were found to be more sensitive. The condition index was also generally higher in summer. Lipid peroxidation, another damage biomarker, was measured in gills to assess if there is any association between the responses of both biomarkers and if they are similarly affected by the same environmental conditions. LPO like DNA damage was higher in summer. This work confirms that DNA damage is a sensitive biomarker to discriminate genotoxic contamination, even in areas considered to suffer low impact from genotoxins.     

Trace Organic Matter in the Ria Formosa,Portugal

The Ria Formosa lagoon, Portugal, receives organic matter from many natural and anthropogenic sources. This paper presents an overview of the contribution these sources made through the use of lipid biomarkers. Sixty-one surface sediment samples were collected and analysed for a range of lipids. Sewage materials were confined to regions within 2 km of discharge points; phytoplankton biomarkers (sterols and fatty acids) suggest production was greatest in the inner parts of the lagoon that also have greatest inorganic nutrient enrichment; terrestrial organic matter was present in relatively low concentrations as shown by both sterol and fatty alcohols; bacteria were widespread although the Sulphate Reducing Bacteria (SRBs, shown by the presence of 3-OH fatty acids) were located near but not adjacent to sewage discharge points.     

Minor and trace elements in the shell of Patella aspera (Röding 1798)

Specimens of the limpet Patella aspera were collected from a clean, coastal marine site and a contaminated estuary on the south coast of Portugal. The shells were analysed individually for their minor (Mg, Sr) and trace element (Fe, Mn, Zn) content. Mean concentrations of these elements in the shell of P. aspera were 4651, 1318, 35.8, 29.9 and 5.5 microg g(-1), respectively. The elemental concentrations exhibited both a marked intra- and inter-population variability. Despite the variability within individual populations, significant differences in the trace element composition were apparent between the shells taken from the two sites. Small shells (< 2 g) provided the best resolution between sites for both manganese and iron. Differences in zinc were best resolved for larger shells. The shell of P. aspera has an extraordinarily high magnesium concentration, which is insensitive to gross salinity differences, and a trace metal assemblage that can be interpreted in terms of environmental exposure. On these grounds, it is recommended that the shell of P. aspera is a tissue for potential use in environmental trace metal monitoring.     

Metallothionein levels in Algerian mice (Mus spretus) exposed to elemental pollution: an ecophysiological approach

The potential use of metallothioneins (MTs) as biomarkers of trace metal contamination was evaluated for the first time in the Algerian mouse (Mus spretus). Mice were collected seasonally in an abandoned mining area (Aljustrel) and in a reference area, both located in southern Portugal. MT levels were quantified in liver and kidney by differential pulse polarography and hepatic elemental concentrations (Mn, Fe, Cu, Zn, Se) were determined by particle-induced X-ray emission. Hepatic iron and selenium concentrations were elevated in mice from Aljustrel mine when compared to reference animals. MTs levels were averagely higher in mice from Aljustrel than those originated from the reference area. A season-dependent significant effect was found on the hepatic and renal MT concentrations, characterized by higher levels in winter and lower in autumn. In contaminated mice positive relationship between liver elemental contents (Cu in autumn and Fe in winter) and MTs were found. The seasonal variation of MT suggests that probably physiological and environmental factors could influence hepatic and renal MT induction. Results seem to imply that some environmental disturbance occur in the vicinity of the Aljustrel mine. Therefore, for the management purposes MT levels should be followed in liver of M. spretus, especially in winter. Furthermore, other physiological factors that could influence MT expression and turnover in Algerian mouse should also be monitored.     

Polycyclic aromatic hydrocarbons in clams Ruditapes decussatus (Linnaeus, 1758)

The concentration of sixteen individual polycyclic aromatic hydrocarbons (PAHs) was measured in the clam Ruditapes decussatus whole soft tissues from several places of the Ria Formosa lagoon (Portugal). Total PAH (tPAH) concentrations were higher in the summer (August) and winter (January) than in the other months and the distribution pattern of the individual PAHs was generally dominated by the 4 aromatic ring PAHs, followed by the 2 + 3 aromatic rings PAHs. Benzo[a]anthracene and acenaphthene were the most representative PAHs of the two fractions. Principal components analysis (PCA) revealed that, in the Ria Formosa, seasonal variations are more important than spatial variations, due to changes in PAH source. These sources ranged from petrogenic to pyrolytic or a mixture of both. The origin of clam PAHs was also assessed by partial least squares (PLS) analysis using nineteen different PAH signatures, taken from the literature. It was possible to identify boat traffic, especially in the summer, as one of the most relevant PAH sources to the Ria Formosa. The influence of boat traffic was revealed by several signatures including diesel soot, oil and weathered oil and a mixture of different individual PAHs usually found in harbour sediments. Other relevant sources included combustion of organic matter such as forest fires and diverse domestic activities, occurring mainly in the summer. Most of the clams were considered safe for human consumption, except for some point samples, which presented unusually high PAH concentrations, suggesting the need for a regular survey of PAHs in clam tissues.     

Metabolic signatures associated with environmental pollution by metals in Doñana National Park using P. clarkii as bioindicator

Bioindicators can reflect the effects of pollutants on their metabolism, being widely used to assess environmental stress. In this sense, the crab Procambarus clarkii has been previously proposed to monitor the contamination in Doñana National Park (southwest Spain) using conventional biomarkers. In this work, a metabolomic approach based on direct infusion mass spectrometry, which allows an easy and quick study of a large number of metabolites in a single run, was used for pollution assessment of this area, considering the biological response of this organism to contamination. In addition, metal accumulation in crab tissues was determined. Thus, the integrated analysis of metabolomic and metallomic data enabled the study of metabolic response of the organism against pollution. Several metabolites were discovered as potential biomarkers of pollution, such as decreased levels of carnosine, alanine, niacinamide, acetoacetate, pantothenic acid, ascorbate, glucose-6-phosphate, arginine, glucose, lactate, phospholipids, and tryglicerides, as well as elevated levels of acetyl carnitine, phosphocholine, choline, and uric acid. In this way, metal-induced toxicity could be related to metabolic impairments, principally oxidative stress, metabolic dysfunction, and dyslipidemia.