Characterization of the cytosolic distribution of priority pollutant metals and metalloids in the digestive gland cytosol of marine mussels: seasonal and spatial variability.

Cytosolic profiles of several priority pollutant metals (Cu, Cd, Zn, Pb) and metalloid As were analyzed in the digestive gland of the mussel (Mytilus galloprovincialis) sampled at locations with different environmental pollution levels along the Croatian coast in the spring and summer season. Size-exclusion chromatography (SEC) connected to inductively coupled plasma mass spectrometry (ICP-MS) was used to determine selected elements bound to cytosolic biomolecules separated based on their molecular size. Copper, cadmium and zinc eluted mostly associated with high molecular weight (HMW) and medium molecular weight (MMW) biomolecules, but with a more prominent elution in the MMW peak at polluted locations which were probably associated with the 20 kDa metallothionein (MT). Elution of all three metals within this peak was also strongly correlated with cytosolic Cd as strong inducer of MT. Lead mostly eluted in HMW biomolecule range, but in elevated cytosolic Pb concentrations, significant amount eluted in low molecular weight (LMW) biomolecules. Arsenic, on the other hand eluted almost completely in LMW range, but we could not distinguish specific molecular weight biomolecules which would be predominant in detoxification mechanism. Seasonal variability in element abundance within specific peaks was present, although not in the same extent, for all elements and locations, especially for As. The results confirm the suitability of the distribution of selected metals/metalloids among different cytosolic ligands as potential indicator for metal exposure. Obtained findings can also serve as guidelines for further separation and characterization of specific cytosolic metal-binding biomolecules.

Assessment of general condition of fish inhabiting a moderately contaminated aquatic environment.

The assessment of general condition of fish in the moderately contaminated aquatic environment was performed on the European chub (Squalius cephalus) caught in September 2009 in the Sutla River in Croatia. Although increases of the contaminants in this river (trace and macro elements, bacteria), as well as physico-chemical changes (decreased oxygen saturation, increased conductivity), were still within the environmentally acceptable limits, their concurrent presence in the river water possibly could have induced stress in aquatic organisms. Several biometric parameters, metallothionein (MT), and total cytosolic protein concentrations in chub liver and gills were determined as indicators of chub condition. Microbiological and parasitological analyses were performed with the aim to evaluate chub predisposition for bacterial bioconcentration and parasitic infections. At upstream river sections with decreased oxygen saturation (~50%), decreased Fulton condition indices were observed (FCI: 0.94 g cm(-3)), whereas gonadosomatic (GSI: 2.4%), hepatosomatic (HSI: 1.31%), and gill indices (1.3%) were increased compared to oxygen rich downstream river sections (dissolved oxygen ~90%; FCI: 1.02 g cm(-3); GSI: 0.6%; HIS: ~1.08%; gill index: 1.0%). Slight increase of MT concentrations in both organs at upstream (gills: 1.67 mg g(-1); liver: 1.63 mg g(-1)) compared to downstream sites (gills: 1.56 mg g(-1); liver: 1.23 mg g(-1)), could not be explained by induction caused by increased metal levels in the river water, but presumably by physiological changes caused by general stress due to low oxygen saturation. In addition, at the sampling site characterized by inorganic and fecal contamination, increased incidence of bacterial bioconcentration in internal organs (liver, spleen, kidney) was observed, as well as decrease of intestinal parasitic infections, which is a common finding for metal-contaminated waters. Based on our results, it could be concluded that even moderate contamination of river water by multiple contaminants could result in unfavourable living conditions and cause detectable stress for aquatic organisms.

Lead concentration increase in the hepatic and gill soluble fractions of European chub (Squalius cephalus)-an indicator of increased Pb exposure from the river water.

PURPOSE: To examine if chronic exposure of feral fish to elevated Pb concentrations in the river water (up to 1 µg L(-1)), which are still lower than European recommendations for dissolved Pb in surface waters (7.2 µg L(-1); EPCEU (Official J L 348:84, 2008)), would result in Pb accumulation in selected fish tissues. METHODS: Lead concentrations were determined by use of HR ICP-MS in the gill and hepatic soluble fractions of European chub (Squalius cephalus) caught in the Sutla River (Croatia-Slovenia). RESULTS: At the site with increased dissolved Pb in the river water, soluble gill Pb levels (17.3 µg L(-1)) were approximately 20 times higher compared to uncontaminated sites (0.85 µg L(-1)), whereas the ratio between contaminated (18.1 µg L(-1)) and uncontaminated sites (1.17 µg L(-1)) was lower for liver (15.5). Physiological variability of basal Pb concentrations in soluble gill and hepatic fractions associated to fish size, condition, sex, or age was not observed, excluding the possibility that Pb increase in chub tissues at contaminated site could be the consequence of studied biotic parameters. However, in both tissues of Pb-exposed specimens, females accumulated somewhat more Pb than males, making female chubs potentially more susceptible to possible toxic effects. CONCLUSIONS: The fact that Pb increase in gill and hepatic soluble fractions of the European chub was not caused by biotic factors and was spatially restricted to one site with increased dissolved Pb concentration in the river water points to the applicability of this parameter as early indicator of Pb exposure in monitoring of natural waters.

Cellular energy allocation in mussels (Mytilus galloprovincialis) from the stratified estuary as a physiological biomarker.

This study demonstrates the usefulness of cellular energy allocation (CEA) evaluations as a physiological biomarker to infer the occurrence of natural stress in native populations of mussels inhabiting the stratified estuary (Krka River estuary, Croatia). Sampling sites were selected based on their differences in the salinity and temperature. The CEA value was calculated as a ratio between available energy (E(a)) and energy consumption (E(c)). Generally, higher values of E(a) were recorded in June than in November, which were especially evident in the storage components (carbohydrates and lipids), while the constitutive component (proteins) remained relatively constant. The highest E(c) was recorded in mussels at estuarine site compared to coastal site, which may be caused by the energetically costly maintenance of osmotic balance. Decrease in CEA recorded in estuarine mussels may ultimately result in a lower amount of energy available for growth, reproduction, or defence against other stresses (e.g. pollution).