Clam Ruditapes Decussatus Research Articles

Some properties of habituation of siphon withdrawal in the slimy clam (Ruditapes decussatus)

A first approximation to the study of learning processes in bivalves is presented. A habituation procedure was developed using the slimy clam Ruditapes decussatus. The percentage of siphon withdrawal when they were exposed to a white light was measured. In Experiment 1, a habituation-discrimination procedure was used to study the stimulus intensity effect (350-lm vs 806 lm). Clams exposed to 350-lm showed a faster habituation than those exposed to 806-lm. Experiment 2 studied the effect of stimulus and intertrial interval (ITI) duration using a 2×2 design. Trials lasted 20 or 180 seconds, and the ITI lasted 5 or 10 minutes. A combined effect of these two parameters was obtained. Habituation was faster in clams exposed to 180-sec trials with a 5-min ITI. Finally, in Experiment 3 clams were trained with five blocks of five trials with a 5-min ITI. Groups differed on trial duration (20 or 180 s). The results showed a general spontaneous recovery effect that was more pronounced for the 180 s group. Also, it was found in this experimental condition a reduced response 24 hours after finishing training (long-term habituation). Altogether, this series of experiments constitutes a first systematic demonstration of habituation in bivalves.

Multiple-biomarker approach in the assessment of bisphenol A effect on the grooved carpet clam Ruditapes decussatus (Linnaeus, 1758)

BackgroundBisphenol A (BPA), a plastic additive monomer, is among the most highly produced chemicals worldwide, and is broadly used in many industries, such as food and beverage containers, milk bottles, and paper products. Previous studies demonstrated that BPA has potential toxicity to aquatic organisms, causing endocrine disturbance and behavioural disorders. The current work aimed to determine the toxic impacts of BPA on the edible marine clam Ruditapes decussatus considering a multi-biomarker approach (mortality, biochemical studies, DNA strand breaks using comet assay, and histopathological examinations with semi-quantitative and quantitative histopathological analyses). The clams were exposed under laboratory conditions to three concentrations of BPA (0 “control”, 1, and 5 µg/L) for a period of 21 days. After the exposure period, BPA impacts were assessed in the digestive gland as a versatile and environmentally relevant organ for ecotoxicological studies.ResultsIn BPA-treated clams, mortality (10%) occurred only at the highest BPA concentration (5 µg/L). Biochemical impairments were detected in a concentration-dependent manner as a consequence of BPA exposure. There were significant increases in malondialdehyde (MDA) and glutathione (GSH) levels, while catalase (CAT) activity was significantly reduced. Our results revealed that BPA induced neurotoxicity in R. decussatus, as evidenced by the inhibition of acetylcholinesterase (AChE) activity in a dose-dependent manner. Furthermore, DNA damage was strongly induced as BPA levels increased. Additionally, our results have been affirmed by alterations in digestive gland tissues at BPA treatments, which consequently can impair the clam’s ability for food absorption; these alterations included mainly atrophic and necrotic digestive tubules, epithelial cell vacuolization, hemocyte infiltration, and intertubular fibrosis. Based on the data obtained from the semi-quantitative and quantitative histopathological analyses, the exposure of the clam’s digestive gland to BPA with concentrations of 1 and 5 µg/L for 21 days showed significant histopathological alterations compared with the control clams.ConclusionThe multi-biomarker approach used in the current study proved to be a useful tool for assessing the impact of diphenylmethane compounds, such as BPA. Water-borne BPA causes oxidative stress, neurotoxicity, genotoxicity, and deleterious effects on the clam digestive gland; all of these could deteriorate clam performance and health, causing tissue dysfunction.

Genetic diversity of the Ruditapes decussatus and evidence of its hybridization with the alien R. philippinarum in the Western Mediterranean Sea

The introduction of alien species in marine ecosystems is often driven by the increasing demand of fishery resources. This is the case of the Manila clam (R. philippinarum), imported in Europe from Japan since the 1970s, to meet the growing demand for clams that the native species, the grooved carpet shell clam (Ruditapes decussatus), could not satisfy. Alien species introduction could threaten the genetic diversity and integrity of the native clam, also causing hybridization (i.e., gene flow from one species into the gene pool of another). Since R. philippinarum recently spread in a few important Mediterranean coastal areas, a combined approach based on morphological characteristics, length differences of two nuclear species-specific markers (ITS2, 5SrDNA) and the sequence of the mitochondrial gene cytochrome c oxidase subunit I (COI), was used to investigate the presence of hybrids in six Mediterranean wetlands (Sardinia, Italy). Eight individuals morphologically identified as R. decussatus were hybrids, having sequences specific to both R. decussatus and R. philippinarum in their nuclear DNA (ITS2 and 5SrDNA). Most of these individuals were found to be post-first generation (F1) hybrids indicating that F1-hybrids may be fertile. Secondly, to study the genetic diversity of R. decussatus in the Sardinian wetlands as well as in its whole distribution area, >380 new COI sequences from the eastern Atlantic Ocean and Mediterranean Sea were analysed along with those available from public databases. Mitochondrial COI data revealed variable haplotype and nucleotide diversities in different areas, which were not dependent on sample sizes. The aquaculture breeding activities and clam transplantation between different countries, along with the long pelagic larval dispersal and the commercial import of other bivalve species might have promoted gene exchange between different sites and thus higher diversity levels in a few wild populations. Our research, evaluating the genetic makeup of wild and hatchery stocks and clarifying the degree of hybridization, can contribute to develop further recommendations for conserving the genetic integrity of R. decussatus.

Genetic Variability and Genetic Differentiation of Populations in the Grooved Carpet Shell Clam (Ruditapes decussatus) Based on Intron Polymorphisms

Genetic Variability and Genetic Differentiation of Populations in the Grooved Carpet Shell Clam (Ruditapes decussatus) Based on Intron Polymorphisms

Probiotics from kefir: Evaluating their immunostimulant and antioxidant potential in the carpet shell clam (Ruditapesdecussatus)

This study aimed to investigate the impact of incorporating kefir into the diet on biometric parameters, as well as the immune and antioxidant responses of the carpet shell clam (Ruditapes decussatus) after an experimental infection by Vibrio alginolyticus. Clams were divided into a control group and a treated group. The control group was fed on spirulina (Arthrospira platensis) alone. While, the treated group was fed on spirulina supplemented with 10% dried kefir. After 21 days, clams were immersed in a suspension of V. alginolyticus 5 × 105 CFU mL −1 for 30 min. Seven days after experimental infection, survival was 100% in both groups. The obtained results showed a slight increase in weight and condition index in clams fed with kefir-supplemented diet for 21 days compared to control clams. Regarding antioxidant responses, the treated group showed higher superoxide dismutase activity compared to the control group. However, the malondialdehyde level was lower in the treated clams than in the control. In terms of immune parameters, the treated group showed slightly elevated activities of phenoloxidase, lysozyme and alkaline phosphatase, whereas a decreased lectin activity was observed compared to the control group. The obtained results suggest that kefir enhanced both the antioxidant and immune response of infected clams.

Determination of hydrophilic marine biotoxins in bivalve molluscs by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)

This work describes a method for the determination of paralytic shellfish toxins in mussels (M. galloprovincialis), oysters (Crassostrea gigas), and clams (Ruditapes decussatus) samples. The acetic acid extraction method is used for simultaneous analysis of Paralytic Shellfish Poisoning (PSP) and the separation of these compounds was achieved on ethylene bridged hybrid (BEH) Amide column with gradient mobile phase composed of acetonitrile (MeCN) and water (as eluent A and eluent B) both containing 2 mM ammonium formate and 50 mM formic acid. The method specificity, linearity, sensitivity, precision, and accuracy were successfully evaluated towards the detection of 13 hydrophilic marine biotoxins analyzed together (single run) in different sample matrixes such as mussels. oysters. and clams. Calibration curves were found to be linear (R2 ≥ 0.98). average Recovery% of 98.5–99.9%. limits of detection (LODs): 0.16–1.92 µg·kg−1; limits of quantitation (LOQs): 0.54–6.41 µg·kg−1) and the relative standard deviation (RSD%) of the precision was ≤ 2.8%. The method was applied for the determination of PSP in shellfish collected from several Tunisian locations. All LC-MS/MS results of analyzed molluscs collected from different Tunisia locations, reveal no PSP toxin group contamination and that the amount found does not exceeding the threshold of 800 µg STXequ·kg−1.

Effect of stocking density on larval performance and post-larvae settlement of clam Ruditapes decussatus (Linnaeus, 1758) in bivalve hatchery production

Larval rearing is a critical phase in bivalve hatchery production and plays a crucial role in the success of spat production. Therefore, understanding the effect of density on larval performance and post-larvae settlement is crucial for optimizing and improving the success of bivalve hatchery production. This study aimed to evaluate the effect of different stocking density on larval performances and settlement rate of clams Ruditapes decussatus. Three different larval rearing densities (10, 20, and 40 larvae/mL) and four settlement densities (35, 70, 100, and 140 larvae/cm²) were studied. The results regarding larval performance showed that high density resulted in low larval survival (18% for D40) compared to low and moderate densities (23% for D10 and 26% for D20, respectively). In terms of post-larvae settlement, a density of 35 larvae/cm² exhibited the highest fixation rate (36%) compared to 33%, 18%, and 9% at 70, 100, and 140 larvae/cm², respectively. Statistical analysis revealed a significant effect of larval density on both survival and growth (F = 11.79, p < 0.05; F = 18.63, p < 0.05, respectively), as well as a highly significant effect on settlement (F = 14.88, p < 0.001). Overall, it is advisable to maintain moderate densities during both larval performance and post-larvae settlement for seed production of R. decussatus in the hatchery.

Hiding from heat: The transcriptomic response of two clam species is modulated by behaviour and habitat

Rising occurrence of extreme warming events are profoundly impacting ecosystems, altering their functioning and services with significant socio-economic consequences. Particularly susceptible to heatwaves are intertidal shellfish beds, located in estuarine areas already stressed by factors such as rainfall events, red tides, eutrophication, and pollution. In Galicia, Northwestern Spain, these beds support vital shellfisheries, featuring the native clam Ruditapes decussatus and the non-indigenous R. philippinarum. Over recent decades, these populations have experienced notable abundance shifts due to various anthropogenic impacts, including climate change. In this habitat, patches of the seagrass Zostera noltei that coexist with bare sand can act as thermal refuges for benthic organisms such as clams. To assess the impact of heatwaves on these ecosystems, a mesocosm experiment was conducted. Juveniles of both clam species in two habitat types—bare sand and sand with Z. noltei—were exposed to simulated atmospheric heatwaves during diurnal low tide for four consecutive days. Subsequent transcriptomic analysis revealed that high temperatures had a more pronounced impact on the transcriptome of R. philippinarum compared to R. decussatus. The habitat type played a crucial role in mitigating heat stress in R. philippinarum, with the presence of Z. noltei notably ameliorating the transcriptomic response. These findings have direct applications in shellfishery management, emphasizing the importance of preserving undisturbed patches of Z. noltei as thermal refuges, contributing to the mitigation of heatwave effects on shellfish populations.

Association between rainfall and Escherichia coli in live bivalve molluscs harvested in Sardinia, Italy

Rainfall is generally accepted as one of the most important factors associated with an increased level of E. coli in bivalve molluscs. Performing microbiological risk assessment is relevant to official control authorities to determine the sanitary status of harvesting areas and, therefore, develop monitoring strategies and identify management practices that could be used to improve the quality and safety of the final product. The present study aimed to investigate the impact of rainfall on the content of E. coli in bivalve molluscs farmed in Sardinia (Italy). Enumeration of E. coli was performed according to the Most Probable Number (MPN) method (ISO 16649-3) on 1,920 bivalve samples collected from 7 regional counties between 2018 and 2020. Bivalve molluscs samples included 955 mussels (Mytilus galloprovincialis), 500 oysters (Crassostrea gigas), 325 clams (Ruditapes decussatus), 94 warty venus (Venus verrucosa), and 46 lagoon cockles (Cerastoderma glaucum). Rainfall data were obtained by the Department of Meteorology of the ARPA Sardegna. For each sampling site, GPS coordinates were used to identify gauge stations within catchment areas. Cumulative rain (mm) was recorded 1, 3, 5, 7, and 15 days before sampling, among which the 7-day cumulative rain was the strongest predictor of E. coli counts. Several thresholds of 7-day cumulative rain (from <10 mm up to >300 mm) before sampling were used to estimate the chances of a non-compliant sample (E. coli levels above the limit for sanitary class A; 230 MPN/100 g). The 7-day cumulative rain was positively associated with the chances of non-compliance. When the 7-day cumulative rain before sampling was >300 mm, 80.5 % of the samples were non-compliant, and the odds of a non-compliant sample were 23.6 times higher, as compared to samples harvested when the 7-day cumulative rainfall was <10 mm. Precipitation data could be a useful tool for interpreting anomalous results from official control authorities and reduce the costs that originate from closure of production areas.

Growth and Physiological Performance in Growth Phenotypes of the Carpet Shell Clam (Ruditapes decussatus) Fed Diets of Variable Lipid/Carbohydrate Ratios

Mixed suspensions of phytoplankton and yeast cells in different proportions were designed to achieve diets with a range of variation (0.6–2.2) in the ratio of lipid to carbohydrate while maintaining protein content constant. Juvenile specimens of the carpet shell clam (Ruditapes decussatus) from two segregated growth phenotypes (fast and slow growers) were food conditioned and the physiological components of the energy balance were determined with these different diet compositions in order to assess the combined effects of endogenous and nutritional factors on growth performance. Conditioning to lipid-rich diets increased growth rate relative to conditioning to carbohydrate-rich diets and resulted in higher scope for growth values in both growth groups. These dietary effects are mainly driven by differences in the absorption efficiency (AE) found between clams fed different food compositions, although the present results do not allow to ascertain whether the reduced AE recorded with the carbohydrate-rich diets results from reduced digestibility of yeasts cells due to structural restrictions or either reflects the digestive imbalance of lipids associated to higher production of metabolic fecal losses. Greater phenotypic plasticity was seen to enable the fast-growing clams fed a carbohydrate-rich diet to overcome the above digestive limitations through an overfeeding response; however, the resource to such kind of physiological mechanism appeared limited by the nutritional conditions (energetic status) prevailing during the conditioning phase.

Impact of polyethylene microplastics on the clam Ruditapes decussatus (Mollusca: Bivalvia): examination of filtration rate, growth, and immunomodulation.

The present study was conducted to assess, for the first time, the effects of a 14 days experimental exposure to polyethylene (PE) based MPs (40-48 µm) on the clam Ruditapes decussatus. Clams were exposed to three different concentrations of MPs in controlled laboratory conditions: 10 µg/L (low), 100 µg/L (medium), and 1000 µg/L (high). The effects of MPs were assessed using a multi-marker approach, including the filtration rate, growth, and the integrity of immune cells (such as haemocyte numbers, viability, and lysosomal membrane destabilization). The results revealed that as the concentration of PE-MPs increased, the filtration rate decreased, indicating that PE-MPs hindered the clams' ability to filter water. Furthermore, there was a noticeable decrease in the overall weight of the clams, particularly in the group exposed to 1000 µg/L. This decrease could be attributed to the impairment of their nutrient filtration function. In terms of immune system biomarkers, exposure to PE-MPs led to immune system disruption, characterized by a significant increase in the number of haemocytic cells, especially in the group exposed to the high concentration. Additionally, there was a notable reduction in the viability of haemocytes, resulting in the destabilization of their lysosomal membranes, particularly in the groups exposed to medium and high PE-MPs concentrations. The findings of this study indicate that the sensitivity of hemolymph parameter changes and filtration rate in R. decussatus exposed to PE-MPs (100 and 1000 µg/L), surpasses that of growth performance and can serve as reliable indicators to assess habitat conditions and contaminant levels.

Are iron ore microparticles toxic for the European clam Ruditapes decussatus? Response elements from biomarker activities and in silico modeling

Inevitably, high concentrations of iron, the most widely produced ore globally, can be found in aquatic environments. To assess the toxicity of iron on aquatic organisms, Ruditapes decussatus specimens were subjected to microparticles derived from two types of iron ore (hematite and magnetite) at four different concentrations (0.5, 1, 1.5, and 5 g/L). The findings revealed that both types of iron ore were absorbed by clams in a concentration-dependent manner. Biomarkers analysis demonstrated significant and organ-specific impacts on the health of the clams caused by these microparticles, which was further supported by computational analyses on bioavailability. Within seven days of exposure, changes were observed in the activities of several enzymes, including catalase, acetylcholinesterase, and glutathione S-transferases, as well as in the rate of lipid peroxidation in both the digestive gland and gills. This study provides an environmental perspective on the toxicological effects of iron ore microparticles.

Assessment of Risks Associated with Extreme Climate Events in Small-Scale Bivalve Fisheries: Conceptual Maps for Decision-Making Based on a Review of Recent Studies

Extreme climate events, such as heatwaves and torrential rain, affect the physiology and functioning of marine species, especially in estuarine habitats, producing severe ecological and socioeconomic impacts when the affected species support important fisheries, such as artisanal shellfisheries. Studies of the impact of sudden decreases in salinity and increases in temperature were reviewed with the aim of producing comprehensive conceptual maps to aid the management of fisheries of the native clams Ruditapes decussatus and Venerupis corrugata, the introduced Ruditapes philippinarum, and the cockle Cerastoderma edule in Galicia (NW Spain). The maps show the effects on mortality, scope for growth, ability to burrow, changes in gonad development or predation risk. While V. corrugata will generally be more affected by low salinity (5 to 15) or high temperature (30 °C) during only two tidal cycles, C. edule populations may recover. Both species are also expected to become more vulnerable to predators. The clams R. philippinarum and R. decussatus will be more resistant, unless extreme events occur after massive spawning episodes; however, the presence of the intertidal seagrass Zostera noltei may buffer the negative effects of high sediment temperature on the growth of some species, such as R. decussatus. Finally, recommendations for assessing climate risk and designing management actions for shellfisheries are given.

Transcriptional Profiling of Populations in the Clam Ruditapes decussatus Suggests Genetically Determined Differentiation in Gene Expression along Parallel Temperature Gradients and between Races of the Atlantic Ocean and West Mediterranean Sea

Ongoing ocean warming due to climate change poses new challenges for marine life and its exploitation. We have used transcriptomics to find genetically based responses to increased temperature in natural populations of the marine clam Ruditapes decussatus, which lives along parallel thermal gradients in southern Europe. Clams of the Atlantic and West Mediterranean races were collected in northern (cool) and a southern (warm) localities. The animals were kept in running seawater in the warm, southern Atlantic locality for a 15-week period. During this period, water temperature was raised to typical southern European summer values. After this period, an expression profile was obtained for a total of 34 clams and 11,025 probes by means of an oligonucleotide microarray. We found distinct transcriptional patterns for each population based on a total of 552 differentially expressed genes (DEGs), indicating innate differences which probably have a genetic basis. Race and latitude contributed significantly to gene expression differences, with very different sets of DEGs. A gene ontology analysis showed that races differed mainly in the genes involved in ribosomal function and protein biosynthesis, while genes related to glutathione metabolism and ATP synthesis in the mitochondria were the most outstanding with respect to north/south transcriptional differences.

Possible Interaction between ZnS Nanoparticles and Phosphonates on Mediterranean Clams Ruditapes decussatus.

This study aims to evaluate the toxicity of ZnS nanoparticles (ZnS NP50 = 50 µg/L and ZnS NP100 = 100 µg/L) and diethyl (3-cyano-1-hydroxy-2-methyl-1-phenylpropyl)phosphonate or P (P50 = 50 µg/L and P100 = 100 µg/L) in the clams Ruditapes decussatus using chemical and biochemical approaches. The results demonstrated that clams accumulate ZnS NPs and other metallic elements following exposure. Moreover, ZnS NPs and P separately lead to ROS overproduction, while a mixture of both contaminants has no effect. In addition, data showed that exposure to P100 resulted in increased levels of oxidative stress enzyme activities catalase (CAT) in the gills and digestive glands. A similar trend was also observed in the digestive glands of clams treated with ZnS100. In contrast, CAT activity was decreased in the gills at the same concentration. Exposure to ZnS100 and P100 separately leads to a decrease in acetylcholinesterase (AChE) levels in both gills and digestive glands. Thus, AChE and CAT after co-exposure to an environmental mixture of nanoparticles (ZnS100) and phosphonate (P100) did not show any differences between treated and non-treated clams. The outcome of this work certifies the use of biomarkers and chemical assay when estimating the effects of phosphonate and nanoparticles as part of an ecotoxicological assessment program. An exceptional focus was given to the interaction between ZnS NPs and P. The antioxidant activity of P has been demonstrated to have an additive effect on metal accumulation and antagonistic agents against oxidative stress in clams treated with ZnS NPs.

Effect of the alveolate parasite Perkinsus olseni infection on sexual maturation and spawning efficiency of the clam Ruditapes decussatus.

The effect of Perkinsus olseni infection on the reproduction ability of clams has been underestimated so far. Although some studies found evidence of reduction of egg production and delay in gonad maturation after infection, the total effect of the infection is still unclear. In this study, Ruditapes decussatus clams from a naïve population were injected with two different doses of P. olseni parasites, a low dose leading to a light infection and a high dose leading to a heavy infection. Clams were maintained during 2 months for maturation, and at the end of the experiment, the spawning was induced, the number of larvae release and mortality were evaluated. During the maturation period, infection level, gonadal stage, condition index, gross biochemical composition and oxidative status of progenitors were evaluated at days 0, 30 and 60 post-injection. The effects of P. olseni infection on clams showed alterations on biochemical parameters, namely lipid peroxidation, a significant mortality and a delayed gonad maturation, with a greater effect in the highly infected individuals. The reproductive capacity of the clams was impaired in both infected groups showing a lower production and a higher mortality rate of larvae. Finally, this study indicates that the production of natural beds with a high prevalence of P. olseni could be compromised by a deregulation of the natural reproduction cycle and a decrease in larvae production by infected animals, probably due to a combination of lower egg production and lower lipid reserves in larvae from infected clams.

Effects of an Endocrine Disruptor Triclosan on Ruditapes decussatus: Multimarker and Histological Approaches.

The aim of this work was to study the ecotoxicological effects of an endocrine disruptor triclosan on the clam Ruditapes decussatus. The bivalves were exposed to three concentrations of this biocide (C1 = 100 ng/L, C2 = 200 ng/L and C3 = 500 ng/L) for three and seven days. The impact was assessed at the gills and digestive glands, through activities of an antioxidant defense biomarker (Gluthatione S-Transferase, GST), a damage biomarker (Malondialdehyde, MDA), and a neurotoxicity biomarker (Acetylcholinesterase, AChE). Furthermore, histological traits were approached in different organs to evaluate any possible alteration induced by triclosan. It appears from this study that both gills and digestive glands responded discernibly to triclosan and effects were concentration-dependent. The stressed clams showed a significant increase in their GST and MDA activities in gills and digestive glands compared to controls for both time slots considered. In turn, the AChE activity was clearly inhibited in both organs in a time dependent way. The histological study made it possible to observe several structural pathologies caused by triclosan in the gills and the digestive gland. These alterations consisted mainly of inflammatory reactions, malformations of the lamellae and fusion of the gill filaments, degeneration of the connective tissue, and the erosion of the gill cilia with the appearance of certain severe alterations (cell necrosis and apoptosis), which can thus cause a malfunction of the gills and eventually lead to a reduction in oxygen consumption and a disruption of the osmoregulation for bivalves. Alterations in the digestive gland have also been detected, mainly by epithelial alterations, thinning of the tubules, and alteration of the basal cell membrane which can impair the ability of clams to absorb food. At germinal cells, several damages were observed in the oocytes which probably disturbed the reproductive function and the fertility of the clams. The damages observed in female gonads were caused by the cytolysis of a large number of oocytes through autophagy and necrosis at 200 ng triclosan/L. Moreover, at 500 ng triclosan/L, hemocytic infiltration was observed in acini and apoptotic bodies reflected in the fragmentation of more than 90% of oocytes.

Physiological Responses of the Bivalves Mytilus galloprovincialis and Ruditapes decussatus Following Exposure to Phenanthrene: Toxicokinetics, Dynamics and Biomarkers Study.

The aim of the current study was to assess the multifaceted effects of the polycylic aromatic hydrocarbon phenanthrene, mainly used in the colouring, explosive, and pharmaceutical industries, on the physiology of two bivalve species with economic value as seafood, namely, the Mediterranean mussel Mytilus galloprovincyalis and the European clam Ruditapes decussatus. The current study assessed how the phenanthrene affected several biomarkers and biometric endpoints in both bivalves, based on an in vivo experiment in silico approach. The bivalves were exposed during four time slots (i.e., 7, 15, 21, and 28 days) to two concentrations of phenanthrene in water (50 µg/L and 100 µg/L). For the clam R. decussatus, an additional contamination of sediment was applied due their typical benthic lifestyle (50 µg/kg and 100 µg/kg). The phenanthrene significantly reduced the ability of bivalves to tolerate desiccation and their Median Lethal Time, and also inhibited the activity of the enzyme acetylcholinesterase in a time-dependent manner. The activity of catalase indicated that bivalves also experienced oxidative stress during the first 21 days of the experiment. The significant decline in catalase activity observed during the last week of the experiment for the mussel M. galloprovincyalis supported a depletion of enzymes caused by the phenanthrene. The phenanthrene has also toxicokinetic and toxicodynamic properties, as assessed by the in silico approach. Overall, the results obtained suggest that the bivalves Ruditapes decussatus and M. galloprovincyalis can be used as a sentinel species in monitoring studies to assess the environmental impact of phenanthene in marine ecosystems. The significance of our findings is based on the fact that in ecotoxicology, little is known about the chronic effects, the simultaneous use of multiple species as bioindicators, and the interactions molecular modelling.

Comparative study of immunological biomarkers in the carpet shell clams (Ruditapes decussatus) from metal-contaminated sites in the South Lagoon of Tunis (Tunisia)

The South Lagoon of Tunis (Tunisia) is a Mediterranean lagoon adversely affected by industrial contaminants, harbour activity and untreated urban sewage. In this lagoon, the clam Ruditapes decussatus has been widely used as a biomonitor of seawater pollution through measurements of parameters related to oxidative stress and neurotoxicity. However, few studies have considered parameters of the immune system of this species in the South Lagoon of Tunis. Therefore, the aim of the present work was to evaluate several immune-related parameters in the cell-free haemolymph of carpet shell clams sampled during August and February from three polluted sites in the South Lagoon of Tunis (S1, S2 and S3) and one less polluted site as a reference site (RS) in order to identify suitable biomarkers for environmental quality assessments of this ecosystem. Concerning the immune-related parameters, seasonal factors modulated phenoloxidase, lysozyme, protease and esterase activity, with lower values measured for samples collected in August than for samples collected in February. In fact, bactericidal activity against two of the pathogenic bacteria tested and the activity of most immune-related enzymes were reduced in the cell-free haemolymph of clams collected from the most sampling sites in August compared to February one. In addition, values of abiotic parameters (temperature, salinity and pH) and metal (cadmium, copper, iron, lead and zinc) concentrations in the clams’ soft tissues, previously obtained and published by the authors, as well as the values of immune-related parameters were integrated using principal component analyses. Results indicated that the values of all measured immune-related parameters were negatively correlated with the temperature values and the variations most of these parameters highlighted that the chemical industrial area (S3) was the most impacted location within the South Lagoon of Tunis. The present study illustrates that the immune-related parameters measured in carpet shell clam cell-free haemolymph represent suitable biomarkers for environmental quality assessments because they provide effective seasonal and spatial discrimination.

Effects of warming on biological interactions between clams and the seagrass Zostera noltei: A case study using open top chambers

Bivalves and seagrasses can interact through diverse environmentally-modulated mechanisms. To assess the effects of climate warming on bivalve-seagrass interactions, we carried out a pioneering field experiment in which open top chambers (OTCs) were used to increase air and sediment temperature in a shellfish bed in NW Spain during two consecutive spring tides (16 days of exposure to OTCs). The temperature increase produced by OTCs was significant, as observed in the daily maximum and mean temperature and in degree hours, although the difference was greater in air and at the sediment surface (up to 8 °C and 3 °C, respectively) than at 5 cm depth (up to 1 °C). Warming was less acute in boxes with the seagrass Zostera noltei, which acted as a thermal buffer, reducing the mean temperature by 1 °C at the sediment surface in OTC boxes and control boxes (without OTCs). Although the short-term increase in temperature did not greatly affect physiological responses of Z. noltei, the carbohydrate reserves and nutrient content increased in the presence of clams. Growth of the native clam Ruditapes decussatus was significantly slower in OTC boxes with bare sediment, and the seagrass thus buffered the negative effect of warming on growth. The presence of Z. noltei may save clams from having to spend energy to burrow deeper to encounter cooler conditions, leaving more energy available for growth. Conversely, growth of the introduced clam R. philippinarum did not differ between habitats or treatments. Our findings confirm a two-way facilitative interaction that may be particularly important in relation to the resilience of both species in the current context of global warming.