Detection of early effects of a single herbicide (diuron) and a mix of herbicides and pharmaceuticals (diuron, isoproturon, ibuprofen) on immunological parameters of Pacific oyster (Crassostrea gigas) spat

In the context of massive summer mortality events of the Pacific oyster Crassostrea gigas, the aim of this study was to investigate the early effects on genes, enzymes and haemocyte parameters implicated in immune defence mechanisms in C. gigas oysters exposed to a potentially hostile environment, i.e. to an herbicide alone or within a mixture. Following 2 h of exposure to the herbicide diuron at 1 μg L⁻¹, the repression of different genes implicated in immune defence mechanisms in the haemocytes and the inhibition of enzyme activities, such as laccase-type phenoloxidase (PO) in the plasma, were observed. The inhibition of superoxide dismutase (SOD) activity in the plasma was also observed after 6 and 24 h of exposure. In the mixture with the herbicides diuron and isoproturon, and the pharmaceutical ibuprofen, catecholase-type PO activity in the plasma and the percentage of phagocytosis in the haemocytes were reduced after 6 h of exposure. Our results showed that early effects on molecular, biochemical and cellular parameters can be detected in the presence of diuron alone or within a mixture, giving an insight of its potential effect in situations that can be found in natural environments, i.e. relatively high concentrations for short periods of time.     

Trade-offs between elimination and detoxification in rainbow trout and common bivalve molluscs exposed to metal stressors

The purpose of this paper was to examine trade-offs between elimination and detoxification in rainbow trout and three common bivalve molluscs (clam, oyster, and scallop) exposed to cadmium (Cd), copper (Cu), and zinc (Zn) based on recent reported experimental data. We incorporated metal influx threshold with subcellular partitioning to estimate rate constants of detoxification (k_d) and elimination (k₂). We found that the relationships between k₂ and k_d were negative for rainbow trout and positive for bivalve molluscs. However, the relationships between k_d and % metal in metabolically detoxified pool were found positive for rainbow trout and negative for bivalve molluscs. Our results also indicated that rainbow trout had higher accumulation (∼60-90%) in metabolically active pool when exposed to essential metals of Cu and Zn and had only 10-50% accumulation in response to non-essential metal of Cd. Based on a cluster analysis, this study indicated that similarity of physiological regulations among study species was found between Cd and Zn. Our study suggested that detoxification can be predicted by an elimination-detoxification scheme with the known elimination rate constant. We concluded that quantification of trade-offs between subcellular partitioning and detoxification provides valuable insights into the ecotoxicology of aquatic organisms and enhances our understanding of the subcellular biology of trace metals.     

Biomarker responsiveness in different tissues of caged Ruditapes philippinarum and its use within an integrated sediment quality assessment

Biomarkers comprising activities of biotransformation enzymes (ethoxyresorufin-O-deethylase -EROD-, dibenzylfluorescein dealkylase -DBF-, glutathione S-transferase -GST), antioxidant enzymes (glutathione reductase -GR- and glutathione peroxidase -GPX), lipid peroxidation -LPO- and DNA strand breaks were analyzed in the clam Ruditapes philippinarum caged at Cádiz Bay, Santander Bay and Las Palmas de Gran Canaria (LPGC) Port (Spain). Sediments were characterized. Digestive gland was the most sensitive tissue to sediment contamination. In Cádiz Bay, changes in LPO regarding day 0 were related with metals. In LPGC Port, DBF, EROD, and GST activity responses suggested the presence of undetermined contaminants which might have led to DNA damage. In Santander Bay, PAHs were related with EROD activity, organic and metal contamination was found to be associated with GR and GST activities and DNA damage presented significant (p < 0.05) induction. R. philippinarum was sensitive to sediment contamination at biochemical level. Biomarkers allowed chemical exposure and sediment quality assessment.     

New metallothionein assay in Scrobicularia plana: heating effect and correlation with other biomarkers

Metallothionein (MT) and other biomarker levels were measured in Scrobicularia plana clams to assess pollution of the Guadalquivir Estuary possibly affected by metals released from Aznalcóllar pyrite mine in 1998. After optimizing reagent concentrations for monobromobimane derivatization, MT levels were quantified by reversed-phase high-performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) in heated or unheated digestive gland extracts and compared to those obtained by differential pulse polarography (DPP). MT content assayed by RP-HPLC-FD in unheated samples was higher than that obtained by DPP and correlated better with metals and anti-oxidant activities. MT assay by RP-HPLC-FD in unheated extracts would be preferable for assessing metal pollution, due to its greater sensitivity and specificity. In addition to MT induction, glyoxalase II inhibition was well correlated with metal contents. Our results suggest that metals at the estuary do not originate from Aznalcóllar spill, but from those carried along by the river and deposited at its concave bank.     

Short Note

This study was aimed to investigate the role of two exotic bivalves ( Ruditapes philippinarum and Musculista senhousia ) on the trophic enrichment and the nutritional quality of sediments from the Sacca di Goro (northern Adriatic Sea) through manipulative experiments. The concentrations of sedimentary total organic matter, biopolymeric carbon, carbohydrates, proteins, lipids, and chlorophyll- a were determined through a 64 days-long period. The experimental setup consisted in three replicated treatments ( R. philippinarum , M. senhousia and simply sand), with treatments and time as fixed factors. All the biochemical classes showed wide fluctuation, and significant differences among treatments were detected during the experimental period. Both bivalve species played a role in sediment enrichment process. At the end of the experimental period, proteins represented the main biochemical class of organic compounds in the treatments with bivalves (48.5 and 44.5% of the biopolymeric fraction in the M. senhousia and R. philippinarum treatments, respectively), while in the control treatment they were only 32.7%. The accumulation of proteic nitrogen increased the nutritional quality of the sedimentary organic matter. Moreover, the presence of M. senhousia also enhanced the accumulation of microphytobenthic carbon.     

Specific biomagnification of polychlorinated dibenzo-p-dioxins and dibenzofurans in tufted ducks (Aythya fuligula), common cormorants (Phalacrocorax carbo) and their prey from Lake Shinji,Japan

Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/DFs) were detected in waterfowl such as common cormorants, tufted ducks, and their prey, namely fish and bivalves from Lake Shinji, Japan. The concentration of total PCDDs/DFs-TEQ was found to be higher in the muscle tissues of common cormorants than in those of tufted ducks. The results of hierarchical cluster analysis implied that the residue distribution pattern of PCDD/DF homologues was considerably different between these two species. Furthermore, biomagnification factors (BMFs) were estimated from bivalves as prey to tufted duck muscles as target organs. Despite the highest concentrations of 1,3,6,8- and 1,3,7,9-TeCDD in tufted ducks and their prey, however, the BMFs of these isomers were calculated to be lower than those of the toxic 2,3,7,8-substituted PCDDs/DFs. On the other hand, log BMF of toxic 2,3,7,8-substituted PCDDs/DFs were significantly higher for lower chlorinated isomers than those of the higher chlorinated isomers. The biota-sediment accumulation factors (BSAFs) of PCDDs/DFs were also estimated using shijimi clam and fish samples against sediment from Lake Shinji. The average BSAFs were estimated and ranged from 4.0×10⁻³ to 2.2×10⁻¹ and 2.0×10⁻⁴ to 2.0×10⁻¹ for bivalve and fish samples, respectively. Based on calculated BMFs and BSAFs, the total PCDD/DF-TEQ levels in the tufted duck were estimated to have been lowest (2.0 pg TEQ/g dry weight basis) in 1947 and highest (9.8 pg TEQ/g) in 1971.     

The distribution of 4-nonylphenol in marine organisms of North American Pacific Coast estuaries

One of the chemical breakdown products of nonylphenol ethoxylates, 4-nonylphenol (4-NP), accumulates in organisms and is of concern as an environmental pollutant due to its endocrine disrupting effects. We measured 4-NP levels in the seawater, sediment, and twelve organisms within the California estuary, Morro Bay, and examined biomagnification of 4-NP using stable isotope abundances (δ¹⁵N and δ¹³C) to quantify trophic position. 4-NP concentrations in organisms from Morro Bay included 25000 ± 8600 ng g⁻¹ lw in liver of California sea lion, 14000 ± 5600 ng g⁻¹ lw in liver of harbor porpoise, 138000 ± 55000 ng g⁻¹ lw in liver of sea otters, 15700 ± 3600 ng g⁻¹ lw in liver of seabirds, 36100 ± 6100 ng g⁻¹ lw in arrow goby fish, 62800 ± 28400 ng g⁻¹ lw in oysters, and 12700 ± 1300 ng g⁻¹ lw in mussels. 4-NP levels generally showed a pattern of trophic dilution among organisms in Morro Bay, with exceptions of biomagnification observed between three trophic links: mussel to sea otter (BMF 10.9), oyster to sea otter (BMF 2.2), and arrow goby to staghorn sculpin (BMF 2.7). Our examination of other west coast estuaries of USA and Canada revealed that mean 4-NP concentrations in gobies and mussels from Morro Bay were significantly higher than those from a more urbanized estuary, San Francisco Bay (goby: 11100 ± 3800 ng g⁻¹ lw) and from a remote estuary, Bamfield Inlet, Canada (goby: 9000 ± 900 ng g⁻¹ lw, mussel: 6100 ± 700 ng g⁻¹ lw). Relative to other estuaries worldwide, 4-NP levels in seawater (0.42 ± 0.16 μg L⁻¹) and sediment (53 ± 14 ng g⁻¹ dw) of Morro Bay are low, but gobies and oysters have higher 4-NP levels than comparable fauna.     

Biosorption of thorium on the external shell surface of bivalve mollusks: the role of shell surface microtopography

External shell surface (ESS) of bivalve mollusks is known to adsorb various metals dissolved in ambient water in high concentration. It is hypothesized here that the surface microtopography of the thin organic coating layer, periostracum, or calcareous shell (if periostracum was destroyed) plays a major role in the adsorption of actinides on ESS. Thorium (natural alpha-emitter) was used in short-term biosorption experiment with shell fragments of five bivalve mollusks. After a 72 h exposure to Th (∼6 kBq L⁻¹), thorium concentration was measured on ESS using laser ablation inductively coupled plasma mass spectrometry; the distribution and density of alpha tracks were subsequently visualized by α-track autoradiography. A trend in reduced Th concentrations on the ESS was observed depending upon the species tested: (group 1 ∼4000 μg g⁻¹) Chlamys islandica (M.), Mercenaria mercenaria (L.), Dreissena polymorpha (P.) > (group 2 ∼1200 μg g⁻¹) Crassostrea virginica (G.) ? (group 3 ∼150 μg g⁻¹) Mytilus edulis L. The microtopography of ESS was characterized by scanning electron microscopy revealing the high porosity of the calcareous surface of C. islandica and M. mercenaria, lamellate surface of periostracum in D. polymorpha, uneven but a weakly porous surface of periostracum of C. virginica, and a nearly smooth surface of the periostracum of M. edulis. This work has demonstrated, for the first time, the presence of a strong correlation between concentration of adsorbed Th and ESS microtopography, and the role of the periostracum in this process is discussed.     

Evaluation of biotargeting and ecotoxicity of Co²⁺-containing nanoscale polymeric complex by applying multi-marker approach in bivalve mollusk Anodonta cygnea

Cobalt (Co²⁺) is present in many nanoscaled materials created for various applications. The key goal of our study was to develop sensitive approaches for assessing the bio-risks associated with using novel Co²⁺-containing nanoscaled polymeric complex (Co-NC). Freshwater bivalve mollusk Anodonta cygnea (Unionidae) was subjected to 14 d action of the developed Co-NC, as well as of Co²⁺ applied in the corresponding concentration (50 μg L⁻¹) or polymeric substance (PS). All experimental groups under study have demonstrated signs of toxic targeting, notably changes in DNA characteristics, oxidative stress (with particularities in each exposed group) and activation of anaerobiosis (Co²⁺ and Co-NC). However, the group exposed to Co-NC showed some advantages that can be related to the activation of metallothionein (MT) function (increase in the level of MT-related SH-groups (MT-SH)): low level of oxyradical formation, no increase in protein carbonylation and vitellogenin-like proteins concentration unlike in Co²⁺ and PS exposed groups. On the other hand, Co²⁺ increased metal (Co, Cu, Zn and Cd) binding to MT (MT-Me) without changes in MT-SH level jointly with activation of oxyradical formation and apoptosis and decreasing of lysosomal membrane stability. PS per se initiated unbalanced changes in activities of the biotransformation enzymes ethoxyresorufin-O-deethylase and glutathione-S-transferase. Thus, Co²⁺ complexing with the developed PS prevented bio-toxic effects of free Co²⁺ ions and PS per se, at least in the studied hydrobiont. The MT-SH was the main distinguishing index of Co-NC group selected by classification and regression tree analysis.     

Bivalve mollusks in metal pollution studies: From bioaccumulation to biomonitoring

Contemporary environmental challenges have emphasized the need to critically assess the use of bivalve mollusks in chemical monitoring (identification and quantification of pollutants) and biomonitoring (estimation of environmental quality). Many authors, however, have considered these approaches within a single context, i.e., as a means of chemical (e.g. metal) monitoring. Bivalves are able to accumulate substantial amounts of metals from ambient water, but evidence for the drastic effects of accumulated metals (e.g. as a TBT-induced shell deformation and imposex) on the health of bivalves has not been documented. Metal bioaccumulation is a key tool in biomonitoring; bioavailability, bioaccumulation, and toxicity of various metals in relation to bivalves are described in some detail including the development of biodynamic metal bioaccumulation model. Measuring metal in the whole-body or the tissue of bivalves themselves does not accurately represent true contamination levels in the environment; these data are critical for our understanding of contaminant trends at sampling sites. Only rarely has metal bioaccumulation been considered in combination with data on metal concentrations in parts of the ecosystem, observation of biomarkers and environmental parameters. Sclerochemistry is in its infancy and cannot be reliably used to provide insights into the pollution history recorded in shells. Alteration processes and mineral crystallization on the inner shell surface are presented here as a perspective tool for environmental studies.