How life history contributes to stress response in the Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>

Introduction  

Within the last decade, numerous studies have investigated the role of environmental history on tolerance to stress of many organisms. This study aims to assess if Manila clams Ruditapes philippinarum may react differently to cadmium exposure and trematode parasite infection (Himasthla elongata) depending on their origin and environmental history in Arcachon Bay (France).     

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.     

Consumption of Ruditapes philippinarum and Ruditapes decussatus: comparison of element accumulation and health risk

Ruditapes philippinarum, a species native from the Indo-Pacific region, was introduced in Europe at the beginning of the 1970s for culture purposes, leading to a massive decrease of the native species Ruditapes decussatus and a high increase of R. philippinarum yields in Europe. Bivalves can accumulate high amounts of metals and thus easily reach concentrations that are toxic not only to themselves but also to consumers. Since differences in the accumulation of pollutants may exist between bivalve species, different health risks may be overcome. For this reason, the level of metals in seafood raises public health concerns, and international organisations like European Food Safety Authority, United States Food and Drug Administration, and Food Standards Australia and New Zealand (FSANZ) set maximum levels (MLs), above which edible seafood cannot be marketed. In order to evaluate the risk associated with the consumption of R. philippinarum and R. decussatus, both clam species were collected in the same site in Ria de Aveiro and the concentration of eight elements determined in organisms before and after a 48-h depuration period. Results evidence that even at low contaminated areas, the MLs for some elements can easily be achieved. The concentrations of As were above the reference values for FSANZ, and the provisional tolerable weekly intake (PTWI) is exceeded for As when more than 0.5 kg of R. decussatus and 0.9 kg of R. philippinarum clam flesh is consumed, in 1 week, by an adult (70 kg). When comparing with other systems worldwide, consumers of depurated clams from this coastal system have a similar or lower risk of exceeding the PTWI for Cd, As, Pb, and Hg. The recently introduced clam, R. philippinarum, accumulates lower amounts of the most health-threatening elements (less than 71 % of Cd, 40 % of As, and 20 % of Hg) than the native R. decussatus, except for Pb. R. philippinarum also reduces more the element burden when subjected to depuration than R. decussatus. Moreover, R. philippinarum allocates a lower proportion of the accumulated elements in the soluble fraction, where they are readily available. Thus, it is safer to consume R. philippinarum than R. decussatus, except when clams come from areas heavily polluted by Pb.     

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.     

Influence of sediment acidification on the bioaccumulation of metals in Ruditapes philippinarum

Background, aim and scope

The influence of pH (range 6.5–8.5) on the uptake of Zn, Cd, Pb, Cu, Ni, Cr, Hg, and As by juveniles of the clam Ruditapes philippinarum was examined in order to understand whether variation in sediment pH has significant repercussions on metal bioaccumulation.

Materials and methods

Clams were exposed to sediments collected in three locations in the Gulf of Cadiz (Huelva, Guadalquivir and Bay of Cadiz) and to contaminated particles derived from an accidental mining spill in Spain.

Results

With a notable exception of metal Cd, the concentration of metals within clams significantly increased (p?<?0.1) when sediment pH was lowered by one or two units. Moreover, the magnitude of this effect was dependent on the type of sediment contamination.

Discussion

Lower pH increases metal solubility and reduces or invert the metal sorption of metals to sediments. Increases in free metal ions in water favors metal uptake by clams, hence pH is an important factor controlling the mobility of these metals within sediments and their subsequent bioaccumulation within biota. Although sediment-water exchange of Cd can increase with acidification, this excess may be counterbalanced by the presence of ligands in seawater preventing the uptake by organism. Besides chlorines, Cd has also an affinity with carbonates and other ligands present in sea water. These Cd-carbonate complexes may reduce the bioavailable to organisms.

Conclusions

These results highlight the potential implications of sediment acidification, either due to the storage excess of organic matter or to the forced capture of CO₂, on the increasing metal availability to benthic organisms.

Recommendations and perspectives

This kind of studies should be increased to address the influence of acidification in the behavior, bioavailability, toxicity, and risk assessment of contaminants associated with sediments either above sub-seabed geological formations in marine environments or in high enriched by organic matter in estuarine areas. Recently, the capture of CO₂ in marine environments has been approved and started; it is necessary to address the potential impacts associated with leakages or other events occurring during the procedure of injection and storage of CO2.     

Interactive effects of cadmium and hypoxia on metabolic responses and bacterial loads of eastern oysters Crassostrea virginica Gmelin

Pollution by toxic metals including cadmium (Cd) and hypoxia are important stressors in estuaries and coastal waters which may interactively affect sessile benthic organisms, such as oysters. We studied metabolic responses to prolonged hypoxic acclimation (2 weeks at 5% O₂) in control and Cd-exposed (30 d at 50 μg L⁻¹ Cd) oysters Crassostrea virginica, and analyzed the effects of these stressors on abundance of Vibrio spp. in oysters. Hypoxia-acclimated oysters retained normal standard metabolic rates (SMR) at 5% O₂, in contrast to a decline of SMR observed during acute hypoxia. However, oysters spent more time actively ventilating in hypoxia than normoxia resulting in enhanced Cd uptake and 2.7-fold higher tissue Cd burdens in hypoxia. Cd exposure led to a significant decrease in tissue glycogen stores, increase in free glucose levels and elevated activity of glycolytic enzymes (hexokinase and aldolase) indicating a greater dependence on carbohydrate catabolism. A compensatory increase in activities of two key mitochondrial enzymes (citrate synthase and cytochrome c oxidase) was found during prolonged hypoxia in control oysters but suppressed in Cd-exposed ones. Cd exposure also resulted in a significant increase in abundance of Vibrio parahaemolyticus and Vibrio vulnificus levels during normoxia and hypoxia, respectively. Overall, Cd- and hypoxia-induced changes in metabolic profile, Cd accumulation and bacterial flora of oysters indicate that these stressors can synergistically impact energy homeostasis, performance and survival of oysters in polluted estuaries and have significant consequences for transfer of Cd and bacterial pathogens to the higher levels of the food chain.     

Mercury accumulation in marine bivalves: influences of biodynamics and feeding niche

Differences in the accumulation of mercury (Hg) in five species of marine bivalves, including scallops Chlamys nobilis, clams Ruditapes philippinarum, oysters Saccostrea cucullata, green mussels Perna viridis, and black mussels Septifer virgatus, were investigated. The bivalves displayed different patterns of Hg accumulation in terms of the body concentrations of methylmercury (MeHg) and total Hg (THg), as well as the ratio of MeHg to THg. Parameters of the biodynamics of the accumulation of Hg(II) and MeHg could reflect the species-dependent Hg concentrations in the bivalves. With the exception of black mussels, we found a significant relationship between the efflux rates of Hg(II) and the THg concentrations in the bivalves. The interspecific variations in the MeHg to THg ratio were largely controlled by the relative difference between the elimination rates of Hg(II) and MeHg. Stable isotope (δ¹³C) analysis indicated that the five bivalve species had contrasting feeding niches, which may also affect the Hg accumulation.     

Relevance of biotic parameters in the assessment of the spatial distribution of gastrointestinal metal and protein levels during spawning period of European chub (Squalius cephalus L.)

The present field study, conducted during the spawning period (April/May) of European chub (Squalius cephalus L.) from the Sava River in Croatia, indicates that seasonal changes of fish physiological state might cause variability in gastrointestinal metal (Cd, Cu, Fe, Mn and Zn), total cytosolic protein and metallothionein (MT) levels. During the period of fish spawning and increased metabolic activity, a significant relationship with chub hepatosomatic index was evident for Fe and Mn in gastrointestinal tissue (r?=?0.35 and 0.26, respectively) and in cytosolic fraction (r?=?0.32 and 0.41, respectively) and for Zn and Fe in the gut content (r?=?0.36 and 0.31, respectively). Total cytosolic protein and MT concentrations followed the same spatial distribution as Fe and Mn in all gastrointestinal fractions and as Zn in the sub-cellular fractions, with higher levels at upstream locations. Due to the role of essential metals in metabolic processes and gonad development, increased feeding and spawning activity in April/May resulted in higher gastrointestinal essential metal (Fe, Mn and Zn) and MT concentrations, which probably follow an increase in Zn concentrations, known as the primary MT inducer. Therefore, biotic factors should be considered as important confounding factors in metal exposure assessment, while their influence on gastrointestinal metal and protein levels should be interpreted depending on the season studied.     

Immunotoxic effects of organotin compounds in Tapes philippinarum

One of the most harmful groups of coastal pollutants is the organotin compounds (OTCs) which have severe effects on both aquatic organisms and mammals including humans. The immunotoxic effects of OTCs were studied in the cultivated clam Tapes philippinarum by determining the immunosuppressant role on in vitro yeast phagocytosis at low doses (0.01, 0.05, 0.1 μM). The phagocytic index was significantly reduced in an irreversible non-lethal manner depending on concentration and lipophilic affinity. The order of inhibition was TBT≥DBT>MBT for butyltins and TPTC>TPTA≥TPTH for triphenyltins.     

Disinfection and removal of human pathogenic bacteria in arctic waste stabilization ponds

Wastewater stabilization ponds (WSPs) are commonly used to treat municipal wastewater in Arctic Canada. The biological treatment in the WSPs is strongly influenced by climatic conditions. Currently, there is limited information about the removal of fecal and pathogenic bacteria during the short cool summer treatment season. With relevance to public health, the objectives of this paper were to determine if treatment in arctic WSPs resulted in the disinfection (i.e., removal of fecal indicator bacteria, Escherichia coli) and removal of selected human bacterial pathogens from the treated effluent. The treatment performance, with focus on microbial removal, was assessed for the one-cell WSP in Pond Inlet (Nunavut [NU]) and two-cell WSP in Clyde River (NU) over three consecutive (2012–2014) summer treatment seasons (late June-early September). The WSPs provided a primary disinfection treatment of the wastewater with a 2–3 Log removal of generic indicator E. coli. The bacterial pathogens Salmonella spp., pathogenic E. coli, and Listeria monocytogenes, but not Campylobacter spp. and Helicobacter pylori, were detected in the untreated and treated wastewater, indicating that human pathogens were not reliably removed. Seasonal and annual variations in temperature significantly (p < 0.05) affected the disinfection efficiency. Improved disinfection and pathogen removal was observed for the two-cell system in Clyde River as compared to the one-cell system in Pond Inlet. A quantitative microbial risk assessment should be performed to determine if the release of low levels of human pathogens into the arctic environment poses a human health risk.     

Combining bioaccumulation and coping mechanism to enhance long-term site-specific risk assessment for zinc susceptibility of bivalves

The purpose of this study is to conduct a long-term site-specific risk assessment for zinc (Zn) susceptibility of bivalves, green mussel Perna viridis and hard clam Ruditapes philippinarum, based on published experimental data by linking the biologically-based damage assessment model with the subcellular partitioning concept. A comprehensive risk modeling framework was developed to predict susceptibility probability of two bivalve species exposed to waterborne Zn. The results indicated that P. viridis accumulates more Zn toxicity, whereas both toxic potency and the recovery rate of Zn are higher for R. philippinarum. We found that negative linear correlations exist in elimination-recovery and elimination-detoxification relationships, whereas a positive linear correlation was observed in recovery-detoxification relationships for bivalves exposed to waterborne Zn. Simulation results showed that the spatial differences of susceptibility primarily resulted from the variation of waterborne Zn concentration under field conditions. We found that R. philippinarum is more susceptible of Zn than P. viridis under the same exposure condition. Results also suggested that Zn posed no significant susceptibility risk to two bivalve species in Taiwan. We suggested that these two species can be used to biomonitor the water quality on Taiwan coastal areas.     

Biochemical effects of acetaminophen in aquatic species: edible clams Venerupis decussata and Venerupis philippinarum

Acetaminophen (paracetamol) is one of the most used pharmaceutical drugs, due to its antipyretic and analgesic properties that turn it into a primary choice in varied pathologies and conditions. However, and despite its massive use, acetaminophen is not exempt of adverse effects, especially when administered in over dosage, which are related to the formation of toxic metabolites by oxidative pathways. It is thus possible to observe that toxicity caused by acetaminophen is usually mediated by reactive oxygen species and can result in multiple effects, ranging from protein denaturation to lipid peroxidation and DNA damage. The occurrence of acetaminophen has been reported in the aquatic environment, being important to address the potential exertion of toxic effects on nontarget environmentally exposed organisms. The present study intended to characterize the effects of acute acetaminophen exposure on physiological traits (antioxidant defense, oxidative damage) of two species of bivalves, namely, the edible clams Venerupis decussata and Venerupis philippinarum. Results showed a significant increase in all oxidative stress biomarkers, evidencing the bioactivation of acetaminophen into a deleterious prooxidant, triggering the onset of deleterious effects. Furthermore, strong interspecific differences were observed among responses of the two tested species, which was a major issue due to intrinsic ecological implications when one considers that both species share the same habitat.     

Metal toxicity in a sediment-dwelling polychaete: Threshold body concentrations or overwhelming accumulation rates?

We followed the net accumulation of As, Cu and Zn in the deposit-feeding polychaete Arenicola marina exposed in the laboratory to natural metal-contaminated sediments, one exposure leading to mass mortality between day 10 and 20, and the other not causing lethality over a period of 60 days of exposure. The worms showed lower total accumulated metal concentrations just before mortality occurred (<20 days) at the lethal exposure, than after 30 days of exposure to sediments not causing mortality. Moreover rates of accumulation of As, Cu and Zn were significantly higher in the lethal exposure than in the sublethal exposure. Our results show that it is not possible to link mortality to a critical total body concentration, and we add to a growing body of literature indicating that metal toxicity occurs when organisms cannot cope with overwhelming influx and subsequent accumulation rates.     

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.     

The effect of nitrate concentration on sulfide-driven autotrophic denitrification in marine sediment

Copepods have been widely used to evaluate toxicity of metals present in marine environments. However, a technical difficulty is to understand the possible routes of metal uptake and to identify in which tissues or organs metals are being accumulated. Traditional techniques are hard to be employed once each organ has to be analyzed separately. Autoradiography is an alternative technique to circumvent this limitation, since metal distribution in tissues can be visualized and quantified, even in small organisms like copepods. In the present study, accumulation and distribution of ⁶⁴Cu in the copepod Calanus hyperboreus was studied using autoradiography. Copepods were exposed for 2 h to copper (2.3 mg L⁻¹; 1.08 MBq ⁶⁴Cu mg⁻¹ Cu) and then allowed to depurate for 2 h in clean seawater. Total ⁶⁴Cu was determined by gamma-spectrometry after a metal exposure and a depuration period. ⁶⁴Cu distribution was determined based on images generated by autoradiography. Metal accumulation was observed on all external surfaces of the copepods, being accumulated mostly on the ventral region, followed by dorsal, urossoma and internal regions. After depuration, radioactivity levels had a decrease in the sum of external body surface. Our results show that copper uptake by C. hyperboreus is fast and that a non-negligible proportion of the accumulated metal can reach internal tissues, which may lead to detrimental physiological effects. Moreover, whole-body autoradiography was demonstrated to be an efficient technique to study copper accumulation and body distribution in a very small organism such as the copepod C. hyperboreus.     

Correlation of foliar MT2b expression with Cd and Zn concentrations in hybrid aspen (Populus tremula × tremuloides) grown in contaminated soil

Metal uptake and its effect on foliar metallothionein 2b (MT2b) mRNA levels were studied in hybrid aspen (Populus tremula × tremuloides) in field conditions. The trees were planted on a site contaminated with several metals, including cadmium (mean 5.1 mg kg⁻¹), chromium (80 mg kg⁻¹), copper (180 mg kg⁻¹), nickel (81 mg kg⁻¹), vanadium (240 mg kg⁻¹) and zinc (520 mg kg⁻¹). Of the ten trace elements analyzed, only Cd and Zn accumulated in the leaves with maximal foliar concentrations of 35 and 2400 mg kg⁻¹ (dry weight), respectively. There was a strong correlation between Cd and Zn concentrations and bioaccumulation factors (concentration in plant/soil) in the leaves, branches and roots, suggesting similar transport mechanisms for these two metals. The levels of MT2b correlated with Cd and Zn concentrations in the leaves, demonstrating that increased MT2b expression is one of the responses of hybrid aspen to chronic metal exposure.     

Antioxidant responses in clam Venerupis philippinarum exposed to environmental pollutant hexabromocyclododecane

The objective of this study was to assess the potential toxic effects of hexabromocyclododecane (HBCD) on tissues of clam Venerupis philippinarum using parameters of antioxidant defenses and oxidative stress. Antioxidant biomarkers including ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), superoxide dismutase (SOD), and glutathione (GSH), as well as DNA damage and lipid peroxidation (LPO) in gills and digestive glands of V. philippinarum, were analyzed after a 1-, 3-, 6-, 10-, and 15-day exposure to seawater containing HBCD at environmentally related concentrations, respectively. The results showed that the activity of most antioxidant enzymes increased, and different trends were detected with exposure time extending. The oxidative stress could be obviously caused in the gills and digestive glands under the experimental conditions. This could provide useful information for toxic risk assessment of environmental pollutant HBCD.     

Changes in cell-type composition in digestive gland of slugs and its influence in biomarkers following transplantation between a relatively unpolluted and a chronically metal-polluted site

Changes in cell-type composition (CCTC) is a general phenomenon that takes place in the digestive gland epithelium of stressed molluscs. The aim of the present work was to determine whether CCTC is a reversible process in the digestive gland of sentinel slugs chronically exposed to metal pollution and how CCTC affects metal accumulation parameters and different cell and tissue biomarkers of exposure and effect. Slugs (Arion ater) from an abandoned zinc mine were transferred to a relatively unpolluted site and the other way around for 3, 10 and 28 d. The volume density of black silver deposits (Vv_BSD) after autometallography, and metallothionein (MT) levels were used as biomarkers of exposure to metals and CCTC and lysosomal responses were selected as effect biomarkers. Results indicated that slugs were sensitive to recent metal pollution; however, slugs chronically exposed to metals presented some characteristic features and were less responsive to pollution cessation without signs of CCTC reversal.     

Accumulation of platinum group elements by the marine gastropod Littorina littorea

The accumulation and trophic transfer of the platinum group elements (PGE): Rh, Pd and Pt; have been studied in short-term (5 day) exposures conducted in aquaria containing the marine macroalga, Ulva lactuca, and/or the grazing mollusc, Littorina littorea. Metals added to sea water (to concentrations of 20 μg L⁻¹) were taken up by U. lactuca in the order Rh, Pt > Pd and by L. littorea in the order Pd ≥ Pt ≥ Rh, with greatest metal accumulation in the latter generally occurring in the visceral complex and kidney. When fed contaminated alga, accumulation of Rh and Pd by L. littorea, relative to total available metal, increased by an order of magnitude, while accumulation of Pt was not readily detected. We conclude that the diet is the most important vector for accumulation of Rh and Pd, while accumulation of Pt appears to proceed mainly from the aqueous phase.