Contamination assessment of a coastal lagoon (Ria de Aveiro, Portugal) using defence and damage biochemical indicators in gill of Liza aurata - An integrated biomarker approach

Fish gill importance in toxicants uptake, bioconcentration and excretion allied to meagre knowledge on branchial damage/protection responses substantiate this study. Five critical sites in Ria de Aveiro (Portugal) were assessed in comparison with a reference site (Torreira), focusing on Liza aurata gill antioxidant defences versus damage (oxidative and genetic). Only in Barra fish displayed damage (lipid peroxidation) though no differences were found in antioxidants. In all other sites, except Rio, antioxidant alterations were found. Thus, fish from Gafanha, Laranjo and Vagos showed higher total glutathione, glutathione peroxidase and catalase. Higher glutathione reductase and glutathione S-transferase activity was also found in the first and the last sites, respectively. In Laranjo, metallothionein levels were higher though lower in Gafanha and Vagos. In general, damage was not accompanied by defences weakening confirming that predicting damage based on antioxidants depletion is not straightforward. The integrated biomarker response index ranked sites as: Gafanha > Barra > Laranjo > Vagos > Rio > Torreira.     

Cadmium accumulation and Cd-binding proteins in marine invertebrates--a radiotracer study

Tissue and subcellular accumulation of cadmium were studied in different tissues of three marine invertebrates (blue mussel Mytilus edulis, the tunicate Ciona intestinalis and the sea star Asterias rubens) using radioactive ¹⁰⁹Cd as a tracer. The organisms were exposed to 0.05, 2 and 50 μg Cd l⁻¹ for 21 days. Quantitative data were obtained by dissecting, weighing and subsequently measuring radioactivity in organs and tissues. Differences between each exposure and each tissue with regard to the amount of radioactivity and metallothionein (MT) content were evaluated. Obvious interspecies differences in Cd accumulation were observed, as well as differences between tissues of the three species. The highest concentrations of Cd in all exposure treatments were found in the hepatopancreas of M. edulis and body wall of A. rubens. Taking all treatments into account, Cd accumulation in the tunic of C. intestinalis was high compared to other tissues from this species. Over 60% of Cd was present in the S50 fraction in all treatments in all three species. Metallothionein levels were increased at the highest Cd-exposure in all species and tissues, except in branchial pharynx of C. intestinalis where the highest MT level was reached following exposure to 2 μg Cd l⁻¹. The most surprising finding was that even the lowest Cd exposure concentration (0.05 μg Cd l⁻¹) caused MT induction in pyloric caeca of A. rubens, but there was no dose-dependent increase in MT at higher exposure levels.     

Roles of regional hydrodynamic and trophic contamination in cadmium bioaccumulation by Pacific oysters in the Marennes-Oléron Bay (France)

The Marennes-Oléron Bay, hosting the largest oyster production in France, is influenced by the historic polymetallic pollution of the Gironde Estuary. Despite management efforts and decreasing emissions in the Gironde watershed, cadmium levels in oysters from the bay are close to the consumption limit (5 μg g⁻¹ dw, EC). From mid April to mid July 2009, we investigated the role of tidal resuspension and regional hydrodynamics on Cd speciation (seawater, SPM, phytoplankton, sediment, microphytobenthos) and bioaccumulation in 18 month-old oysters (gills, digestive glands, rests of tissues) reared under natural conditions (i) at ∼60 cm above the sediment and (ii) on the sediment surface. Dissolved and particulate Cd concentrations in surface and bottom waters were similar and constant over tidal cycle suggesting the absence of Cd release during sediment resuspension. Temporal dissolved and particulate Cd concentrations were closely related to Gironde Estuary water discharges, showing increasing concentrations during flood situations and decreasing concentrations afterwards. Cd depletion in the water column was associated with increasing Cd in the [20-100 μm] plankton fraction, suggesting Cd bioaccumulation. After 3 months, enrichment factors of Cd in tissues of oysters exposed in the water column and directly on the sediment were respectively 3.0 and 2.2 in gills, 4.7 and 3.2 in digestive glands and 4.9 and 3.4 in remaining tissues. Increasing Cd bioaccumulation in gills, digestive glands and remaining tissues can be related to elevated dissolved Cd in the bay, suggesting gill contamination via the direct pathway and subsequent internal redistribution of Cd to other organs and tissues. Elevated Cd contents in oysters reared on tables could be attributed to different trophic Cd transfer (phytoplankton versus microphytobenthos) or to different oyster metabolisms between the rearing conditions as suggested by metallothionein concentrations.     

Toxicity of silver nanoparticles to rainbow trout: A toxicogenomic approach

Silver (Ag) nanoparticles are used as antimicrobial adjuvant in various products such as clothes and medical devices where the release of nano-Ag could contaminate the environment and harm wildlife. The purpose of this study was to examine the sublethal effects of nano-Ag and dissolved Ag on Oncorhynchus mykiss rainbow trout. Hepatic Ag contents and changes in gene expression were monitored to provide insights on bioavailability and mode of action of both forms of silver. Fish were exposed to increasing concentrations (0.06, 0.6 and 6 μg L⁻¹) of nano-Ag (20 nm) and silver nitrate (AgNO₃) for 96 h at 15 °C. A gene expression analysis was performed in the liver using a DNA microarray of 207 stress-related genes followed by a quantitative polymerase chain reaction on a selection of genes for validation. The biochemical markers consisted of the determination of labile zinc, metallothioneins, DNA strand breaks, lipid peroxidation (LPO) and vitellogenin-like proteins. The analysis of total Ag in the aquarium water revealed that nano-Ag was mostly aggregated, with 1% of the total Ag being dissolved. Nevertheless, hepatic Ag content was significantly increased in exposed fish. Indeed, dissolved Ag was significantly more bioavailable than nano-Ag only at the highest concentration with 38 ± 10 and 11 ± 3 ng Ag mg⁻¹ proteins for dissolved and nano-Ag respectively. Exposure to both forms of Ag led to significant changes in gene expression for 13% of tested gene targets. About 12% of genes responded specifically to nano-Ag, while 10% of total gene targets responded specifically to dissolved Ag. The levels of vitellogenin-like proteins and DNA strand breaks were significantly reduced by both forms of Ag, but DNA break levels were lower with nano-Ag and could not be explained by the presence of ionic Ag. Labile zinc and the oxidized fraction of metallothioneins were increased by both forms of Ag, but LPO was significantly induced by nano-Ag only. A discriminant function analysis revealed that the responses obtained by biochemical markers and a selection of ten target genes were able to discriminate completely (100%) the effects of both forms of Ag. Exposure to nano-Ag involved genes in inflammation and dissolved Ag involved oxidative stress and protein stability. Hence, the toxicity of Ag will differ depending on the presence of Ag nanoparticles and aggregates.     

Oxidative stress on land snail Helix aspersa as a sentinel organism for ecotoxicological effects of urban pollution with heavy metals

The oxidative stress in the digestive gland of the land snail Helix aspersa was considered as a bioindicator for atmospheric pollution with heavy metals from several industries and vehicular traffic in Kafr El-Zayat city. Regional means of heavy metals concentration of all sites were 0.71, 7.09, 0.71, 2.68, 41.44 and 18.01 mg kg⁻¹ wet mass for Cd, Mn, Ni, Pb, Zn and Cu, respectively. In addition, the highest values of Cd concentrations were found 1.22 and 1.73 mg kg⁻¹ wet mass in S1 (Potato International Center) and S4 (The Nile bank), respectively. Lactate dehydrogenase (D-LDH(and recorded lipid peroxidation (LPO) levels were significantly high in S1 and S2 (Traffic station). On the other hand, the highest activity of catalase (CAT) was found in S2 (194.04% of control), while the activity of glutathione peroxidase (GP_x) reached the highest significant value in S1.     

Biological Adverse Effects on Bivalves Associated with Trace Metals Under Estuarine Environments

Toxic effects of pollutants on marine organisms can be studied both by performing field measurements, and by undertaking laboratory simulation experiments. Here is described the effect of trace metals Zn, Cd, Pb and Cu on the clam Scrobicularia plana along a salinity gradient simulated in a hypothetical estuary using simulation experiments. The simulator produces a continuous entry of trace metals into the estuary through injection in the lower salinity tank of the system. The clams were exposed during two weeks to different concentration of trace metals to assess the bioaccumulation process along a salinity gradient. Bivalves were analysed for body tissue residue to determine the bioaccumulation factors related to each metal and the salinity influence was addressed. Differences among tanks were observed as a result of the salinity gradient. In the achieved assays, the mechanism of bioaccumulation of Zn and Cd in organisms was more efficient at high salinity values. Bioaccumulation factors for both metals showed a linear increase with the increase of salinity values. It seems that the mechanism of bioaccumulation of Pb and Cu in organisms was dependent on two simultaneous processes: the proximity to the input point of metals and the low salinity values.     

Impact of mercury on the activity pattern of a marker enzyme in a freshwater bivalve

Sessile benthic organisms are universally accepted as bio-indicators of environmental pollution and lysosomal membrane is often the target of injury by xenobiotics resulting in destabilization. Effect of mercury on the activity pattern of the marker enzyme, Acid Phosphatase in the gill and hepatopancreas of the freshwater mussel, Lamellidens corrianus (Lea) at 24, 72, and 168 h, post-exposure discussed. ACP activity was determined as given in Sigma Technical Bulletin (No. 104) and μmol of p-nitrophenol liberated/mg protein/h calculated. The work aimed to study the duration of exposure and concentration of the metal influence, the direction of the response of the enzyme in the two tissues studied and results show that the concentration of the metal as well as the period of exposure do influence the enzyme activity and may have predictive value as a biomarker of impending population changes. Higher concentration of the metal is assumed to have induced stress proteins like metallothioneins.     

Metallothioneins failed to reflect mercury external levels of exposure and bioaccumulation in marine fish--considerations on tissue and species specific responses

The suitability of metallothioneins (MT) in fish as biomarker of exposure to mercury has been questioned. Therefore, this study aimed at investigating the relationship between external levels of exposure, mercury accumulation and MT content, assessing species and tissue specificities. Two ecologically different fish species - Dicentrarchus labrax and Liza aurata - were surveyed in an estuary historically affected by mercury discharges. Total mercury (T-Hg) and MT content were determined in gills, blood, liver, kidney, muscle and brain. All tissues reflected differences in T-Hg accumulation in both species, although D. labrax accumulated higher levels. Regarding MT, D. labrax revealed a depletion in brain MT content and an incapacity to induce MT synthesis in all the other tissues, whereas L. aurata showed the ability to increase MT in liver and muscle. Tissue-specificities were exhibited in the MT inducing potential and in the susceptibility to MT decrease. L. aurata results presented muscle as the most responsive tissue. None of the investigated tissues displayed significant correlations between T-Hg and MT levels. Overall, the applicability of MT content in fish tissues as biomarker of exposure to mercury was uncertain, reporting limitations in reflecting the metal exposure levels and the subsequent accumulation extent.     

Assessment of biological effects of environmental pollution along the NW Mediterranean Sea using red mullets as sentinel organisms

A biomonitoring program was carried out in spring and autumn in three pollution hot-spots and sensitive areas of the NW Mediterranean Sea using red mullets (Mullus barbatus) as sentinel organisms and a battery of biomarkers together with gonad histology. In fish from anthropogenic impacted areas (Fos-sur-mer, Cortiou, Arenzano, Delta of Ebro) lysosomal membrane destabilization occurred indicating disturbed health. There were no significant differences in metallothionein (MT) levels among stations. Peroxisomal acyl-CoA oxidase (AOX) activity was highest in fish from Cortiou. Both MT levels and AOX activities were significantly correlated with gamete development. Prevalence of melanomacrophage centers were high in Cortiou in all samplings and in Fos-sur-mer in September samplings. In conclusion, the application of a battery of biomarkers in red mullets provided relevant data for the assessment of environmental pollution in the NW Mediterranean Sea but also showed the difficulties of using native fish as sentinels. For future studies caging strategies are recommended.