Symbiont photosynthesis in giant clams is promoted by V-type H+-ATPase from host cells.

Giant clams (genus Tridacna) are the largest living bivalves and, like reef-building corals, host symbiotic dinoflagellate algae (Symbiodinium) that significantly contribute to their energy budget. In turn, Symbiodinium rely on the host to supply inorganic carbon (Ci) for photosynthesis. In corals, host 'proton pump' vacuolar-type H+-ATPase (VHA) is part of a carbon-concentrating mechanism (CCM) that promotes Symbiodinium photosynthesis. Here, we report that VHA in the small giant clam (Tridacna maxima) similarly promotes Symbiodinium photosynthesis. VHA was abundantly expressed in the apical membrane of epithelial cells of T. maxima's siphonal mantle tubule system, which harbors Symbiodinium Furthermore, application of the highly specific pharmacological VHA inhibitors bafilomycin A1 and concanamycin A significantly reduced photosynthetic O2 production by ∼40%. Together with our observation that exposure to light increased holobiont aerobic metabolism ∼5-fold, and earlier estimates that translocated fixed carbon exceeds metabolic demand, we conclude that VHA activity in the siphonal mantle confers strong energetic benefits to the host clam through increased supply of Ci to algal symbionts and subsequent photosynthetic activity. The convergent role of VHA in promoting Symbiodinium photosynthesis in the giant clam siphonal mantle tubule system and coral symbiosome suggests that VHA-driven CCM is a common exaptation in marine photosymbioses that deserves further investigation in other taxa.

Trematode infection modulates cockles biochemical response to climate change.

Resulting mainly from atmospheric carbon dioxide (CO2) build-up, seawater temperature rise is among the most important climate change related factors affecting costal marine ecosystems. Global warming will have implications on the water cycle, increasing the risk of heavy rainfalls and consequent freshwater input into the oceans but also increasing the frequency of extreme drought periods with consequent salinity increase. For Europe, by the end of the century, projections describe an increase of CO2 concentration up to 1120 ppm (corresponding to 0.5 pH unit decrease), an increase in the water temperature up to 4 °C and a higher frequency of heavy precipitation. These changes are likely to impact many biotic interactions, including host-parasite relationships which are particularly dependent on abiotic conditions. In the present study, we tested the hypothesis that the edible cockle, Cerastoderma edule, exposed to different salinity, temperature and pH levels as proxy for climate change, modify the infection success of the trematode parasite Himasthla elongata, with consequences to cockles biochemical performance. The results showed that the cercariae infection success increased with acidification but higher biochemical alterations were observed in infected cockles exposed to all abiotic experimental stressful conditions tested. The present study suggested that changes forecasted by many models may promote the proliferation of the parasites infective stages in many ecosystems leading to enhanced transmission, especially on temperate regions, that will influence the geographical distribution of some diseases and, probably, the survival capacity of infected bivalves.

Digestive selection underlies differential utilization of phytoplankton and sedimentary organics by infaunal bivalves: Experiments with cockles (Cerastoderma edule) using cross-labelled mixed diets.

Differential utilization of phytoplankton and detrital particles present in natural sediments of mud-flats was studied in a series of experiments performed on the infaunal bivalve Cerastoderma edule. In order to assess digestive selection, parameters of food processing (organic ingestion rate: OIR, gross absorption efficiency: GAE and gut passage time: GPT) were recorded for each organic component in different combinations of food particles radio-labelled with (14)C. Experimental design included the use of both labelled diets of a sole organic component and cross-labelled diets; i.e., mixed suspensions presenting alternatively labelled one of the various components tested: phytoplankton cells, sedimentary organic particles and particulate detritus from vascular salt-marsh plants. Preferential absorption of phytoplankton was accounted for by absorption efficiency values that were two-fold those for sedimentary detritus when recorded with mixed diets of both organic components. Two factors contributed to this difference: a) higher digestibility of microalgae, measured as the ratio of GAE to GPT, and b) faster gut passage of detrital particles that results from digestive selection likely involving the preferential incorporation of phytoplankton into the digestive gland. However, when diets based on a sole organic component (either phytoplankton or detritus) were compared, larger GPT were recorded for detrital particles that enabled improving GAE of this rather refractory food. Overall results of these experiments are consistent with most studies in trophic ecology based on stable isotopes enrichment, concerning both the diversity of trophic sources used by marine bivalves and its preferential utilization of phytoplankton over phyto-detritus.

Impact of high pCO2 on shell structure of the bivalve Cerastoderma edule.

Raised atmospheric emissions of carbon dioxide (CO2) result in an increased ocean pCO2 level and decreased carbonate saturation state. Ocean acidification potentially represents a major threat to calcifying organisms, specifically mollusks. The present study focuses on the impact of elevated pCO2 on shell microstructural and mechanical properties of the bivalve Cerastoderma edule. The mollusks were collected from the Baltic Sea and kept in flow-through systems at six different pCO2 levels from 900 µatm (control) to 24,400 µatm. Extreme pCO2 levels were used to determine the effects of potential leaks from the carbon capture and sequestration sites where CO2 is stored in sub-seabed geological formations. Two approaches were combined to determine the effects of the acidified conditions: (1) Shell microstructures and dissolution damage were analyzed using scanning electron microscopy (SEM) and (2) shell hardness was tested using nanoindentation. Microstructures of specimens reared at different pCO2 levels do not show significant changes in their size and shape. Likewise, the increase of pCO2 does not affect shell hardness. However, dissolution of ontogenetically younger portions of the shell becomes more severe with the increase of pCO2. Irrespective of pCO2, strong negative correlations exist between microstructure size and shell mechanics. An additional sample from the North Sea revealed the same microstructural-mechanical interdependency as the shells from the Baltic Sea. Our findings suggest that the skeletal structure of C. edule is not intensely influenced by pCO2 variations. Furthermore, our study indicates that naturally occurring shell mechanical property depends on the shell architecture at µm-scale.

Understanding spatial distributions: negative density-dependence in prey causes predators to trade-off prey quantity with quality.

Negative density-dependence is generally studied within a single trophic level, thereby neglecting its effect on higher trophic levels. The 'functional response' couples a predator's intake rate to prey density. Most widespread is a type II functional response, where intake rate increases asymptotically with prey density; this predicts the highest predator densities at the highest prey densities. In one of the most stringent tests of this generality to date, we measured density and quality of bivalve prey (edible cockles Cerastoderma edule) across 50 km² of mudflat, and simultaneously, with a novel time-of-arrival methodology, tracked their avian predators (red knots Calidris canutus). Because of negative density-dependence in the individual quality of cockles, the predicted energy intake rates of red knots declined at high prey densities (a type IV, rather than a type II functional response). Resource-selection modelling revealed that red knots indeed selected areas of intermediate cockle densities where energy intake rates were maximized given their phenotype-specific digestive constraints (as indicated by gizzard mass). Because negative density-dependence is common, we question the current consensus and suggest that predators commonly maximize their energy intake rates at intermediate prey densities. Prey density alone may thus poorly predict intake rates, carrying capacity and spatial distributions of predators.

Propensity to metal accumulation and oxidative stress responses of two benthic species (Cerastoderma edule and Nephtys hombergii): are tolerance processes limiting their responsiveness?

The chronic exposure of benthic organisms to metals in sediments can lead to the development of tolerance mechanisms, thus diminishing their responsiveness. This study aims to evaluate the accumulation profiles of V, Cr, Co, Ni, As, Cd, Pb and Hg and antioxidant system responses of two benthic organisms (Cerastoderma edule, Bivalvia; Nephtys hombergii, Polychaeta). This approach will provide clarifications about the ability of each species to signalise metal contamination. Organisms of both species were collected at the Tagus estuary, in two sites with distinct contamination degrees (ALC, slightly contaminated; BAR, highly contaminated). Accordingly, C. edule accumulated higher concentrations of As, Pb and Hg at BAR compared to ALC. However, antioxidant responses of C. edule were almost unaltered at BAR and no peroxidative damage occurred, suggesting adjustment mechanisms to the presence of metals. In contrast, N. hombergii showed a minor propensity to metal accumulation, only signalising spatial differences for As and Pb and accumulating lower concentrations of metals than C. edule. The differences in metal accumulation observed between species might be due to their distinctive foraging behaviour and/or the ability of N. hombergii to minimise the metal uptake. Despite that, the accumulation of As and Pb was on the basis of the polychaete antioxidant defences inhibition at BAR, including CAT, SOD, GR and GPx. The integrated biomarker response index (IBRv2) confirmed that N. hombergii was more affected by metal exposure than C. edule. In the light of current findings, in field-based studies, the information of C. edule as a bioindicator should be complemented by that provided by another benthic species, since tolerance mechanisms to metals can hinder a correct diagnosis of sediment contamination and of the system's health. Overall, the present study contributed to improve the lack of fundamental knowledge of two widespread and common estuarine species, providing insights of the metal accumulation profiles under a scenario of chronic contamination. Finally, this work provided useful information that can be applied in the interpretation of future environmental monitoring studies.

Long-term epidemiological study of disseminated neoplasia of cockles in Galicia (NW Spain): temporal patterns at individual and population levels, influence of environmental and cockle-based factors and lethality.

The dynamics of disseminated neoplasia (DN) affecting cockles Cerastoderma edule (L.) in Galicia was addressed at individual and population levels. Early stage of DN was characterized by isolated neoplastic cells occurring in branchial vessels or in the connective tissue of gills, mantle, gonad or digestive gland. As disease progressed, the neoplastic cells appeared loose in foci and became widely distributed throughout the organs. In advanced stages, the connective tissue of most organs was infiltrated by neoplastic cells, which displaced normal cells, leading to the loss of the normal tissue/organ architecture. Host defence reaction was occasionally observed. A field survey performed for 7 years, in two cockle beds located in different Galician Rías, showed that DN is a hyperendemic disease usually present all year-round at high prevalence in adult cockles but with annual prevalence minima in spring likely due to the death of heavily affected cockles, concurrently with gonad ripeness-spawning. DN was detected in the cockles ranging from 10 to 39 mm in size; the highest DN prevalence and severity corresponded to the cockles of intermediate size/age (22-29 mm/0.7-1 year old). Sex did not appear to influence DN occurrence. An inhibitory effect of DN on cockle gametogenesis was detected.

Natural selection by pulsed predation: survival of the thickest.

Selective predation can lead to natural selection in prey populations and may alleviate competition among surviving individuals. The processes of selection and competition can have substantial effects on prey population dynamics, but are rarely studied simultaneously. Moreover, field studies of predator-induced short-term selection pressures on prey populations are scarce. Here we report measurements of density dependence in body composition in a bivalve prey (edible cockle, Cerastoderma edule) during bouts of intense predation by an avian predator (Red Knot, Calidris canutus). We measured densities, patchiness, morphology, and body composition (shell and flesh mass) of cockles in a quasi-experimental setting, i.e., before and after predation in three similar plots of 1 ha each, two of which experienced predation, and one of which remained unvisited in the course of the short study period and served as a reference. An individual's shell and flesh mass declined with cockle density (negative density dependence). Before predation, cockles were patchily distributed. After predation, during which densities were reduced by 78% (from 232 to 50 cockles/m2), the patchiness was substantially reduced, i.e., the spatial distribution was homogenized. Red Knots selected juvenile cockles with an average length of 6.9 ± 1.0 mm (mean ± SD). Cockles surviving predation had heavier shells than before predation (an increase of 21.5 percentage points), but similar flesh masses. By contrast, in the reference plot shell mass did not differ statistically between initial and final sampling occasions, while flesh mass was larger (an increase of 13.2 percentage points). In this field study, we show that Red Knots imposed a strong selection pressure on cockles to grow fast with thick shells and little flesh mass, with selection gradients among the highest reported in the literature.

Potential use of fatty acid profiles of the adductor muscle of cockles (Cerastoderma edule) for traceability of collection site.

Geographic traceability of seafood is key for controlling its quality and safeguarding consumers' interest. The present study assessed if the fatty acid (FA) profile of the adductor muscle (AM) of fresh cockles (Cerastoderma edule) can be used to discriminate the origin of specimens collected in different bivalve capture/production areas legally defined within a coastal lagoon. Results suggest that this biochemical approach holds the potential to trace sampling locations with a spatial resolution <10 Km, even for areas with identical classification for bivalve production. Cockles further away from the inlet, i.e. in areas exposed to a higher saline variation, exhibited lower levels of saturated fatty acids, which are key for stabilizing the bilayer structure of cell membranes, and a higher percentage of polyunsaturated fatty acids, which enhance bilayer fluidity. Results suggest that the structural nature of the lipids present in the AM provides a stable fatty acid signature and holds potential for tracing the origin of bivalves to their capture/production areas.

The reproductive cycle, condition index, and glycogen reserves of the cockles Cerastoderma glaucum from the Gulf of Gabès (Tunisia).

The gametogenic cycle of the Cerastoderma glaucum was analyzed using both qualitative and semi-quantitative methods. The condition index and glycogen concentrations were determined in order to provide information on energy storage. The cockles were collected monthly from a Bayyadha site located 15 km south of Sfax City (Gulf of Gabès) between January 2007 and January 2008. From histological point of view, we applied two approaches: (i) the qualitative method describing the various stages of gamete development for males and females during a cycle of 13 months, and (ii) the semi-quantitative method concerning the estimation of different tissue surfaces. The results showed that there is evidence of three periods of reproduction in this population. A comparison between the surfaces occupied by the three organs showed that the foot and the gonad surfaces are higher than the surface of the adductor muscle. This could suggest that these two organs are more involved in the process of glycogen reserve storage. The results of the glycogen concentrations in the different tissues (gonad, adductor muscle, and "remainders") show that during the second and third periods of reproduction, glycogen was stored in the adductor muscle and in the remainder during sexual rest, and in the gonad during the gametogenesis phases in order to supply the reproductive effort. On the contrary, in the first period of reproduction, the low concentrations of glycogen recorded in the gonad coincided with its high degree of development. This fact could be related to environmental conditions (low temperature and food) recorded during this period.

Transcriptional response of stress-regulated genes to cadmium exposure in the cockle Cerastoderma glaucum from the gulf of Gabès area (Tunisia).

This study investigates cadmium effects on key messenger RNA (mRNA) expression (MT, MnSOD, CuZnSOD, CAT, ABCB1, HSP70, and CO1) by qPCR in the cockle Cerastoderma glaucum after chronic exposure to two high but environmentally relevant concentrations of CdCl2 (50 µg/L and 5 mg/L) for 12 h to 18 days. Cd accumulation measured in cockles' tissues is significantly higher in both treatment conditions compared to controls and in a dose-dependent manner. Stress on stress tests performed at different times of the experiment clearly demonstrated that exposure to both concentrations of Cd significantly affects cockle survival time in air. Important changes in gene transcription were also highlighted. In particular, MT, HSP70, CAT, and CuZnSOD seem to be relevant biomarkers of Cd exposure because (1) their mRNA levels increase upon exposure and (2) they are highly correlated to Cd accumulation in tissues. Results may be useful for control strategies and for the use of cockles as sentinel organisms.

Interactions between seagrass complexity, hydrodynamic flow and biomixing alter food availability for associated filter-feeding organisms.

Seagrass shoots interact with hydrodynamic forces and thereby a positively or negatively influence the survival of associated species. The modification of these forces indirectly alters the physical transport and flux of edible particles within seagrass meadows, which will influence the growth and survivorship of associated filter-feeding organisms. The present work contributes to gaining insight into the mechanisms controlling the availability of resources for filter feeders inhabiting seagrass canopies, both from physical (influenced by seagrass density and patchiness) and biological (regulated by filter feeder density) perspectives. A factorial experiment was conducted in a large racetrack flume, which combined changes in hydrodynamic conditions, chlorophyll a concentration in the water and food intake rate (FIR) in a model active filter-feeding organism (the cockle). Results showed that seagrass density and patchiness modified both hydrodynamic forces and availability of resources for filter feeders. Chlorophyll a water content decreased to 50% of the initial value when densities of both seagrass shoots and cockles were high. Also, filter feeder density controlled resource availability within seagrass patches, depending on its spatial position within the racetrack flume. Under high density of filter-feeding organisms, chlorophyll a levels were lower between patches. This suggests that the pumping activity of cockles (i.e. biomixing) is an emergent key factor affecting both resource availability and FIR for filter feeders in dense canopies. Applying our results to natural conditions, we suggest the existence of a direct correlation between habitat complexity (i.e. shoot density and degree of patchiness) and filter feeders density. Fragmented and low-density patches seem to offer both greater protection from hydrodynamic forces and higher resource availability. In denser patches, however, resources are allocated mostly within the canopy, which would benefit filter feeders if they occurred at low densities, but would be limiting when filter feeder were at high densities.

Ingestion and absorption of particles derived from different macrophyta in the cockle Cerastoderma edule: effects of food ration.

We analyzed the capacity of the common cockle Cerastoderma edule to utilize detrital food particles obtained from three different macrophytes: the vascular plant Juncus maritimus and two green macroalgae (Ulva lactuca and Enteromorpha sp.). We measured feeding and digestive parameters at three concentrations of detritus (0.5, 1.0 and 3.0 mm(3) l(-1)), so that functional relationships between ingestive and digestive processes could be assessed. Increasing concentrations of detritus (food) resulted in a reduction in filtering activity (clearance rate l h(-1)), but an increase in ingestion rate. Consequently, gut content also increased with increasing food concentration, irrespective of food type. In contrast, the trend followed by absorption efficiency with increasing ingestion rate was determined by food type, being significantly reduced (from 0.63 to 0.11) with Juncus but remaining almost constant with the green macroalgae (0.58 ± 0.07 with Ulva) or only minimally reduced (from 0.66 to 0.48 with Enteromorpha). This differential response had clear consequences for energy uptake: absorption rate increased with increasing particulate organic matter with Enteromorpha but decreased with Juncus. We discuss the possible role of digestive parameters such as digestibility, gut content and gut-residence time in the differential utilization of detrital matter from different vegetal origins by cockles.

Cross-habitat interactions among bivalve species control community structure on intertidal flats.

Increasing evidence shows that spatial interactions between sedentary organisms can structure communities and promote landscape complexity in many ecosystems. Here we tested the hypothesis that reef-forming mussels (Mytilus edulis L.), a dominant intertidal ecosystem engineer in the Wadden Sea, promote abundances of the burrowing bivalve Cerastoderma edule L. (cockle) in neighboring habitats at relatively long distances coastward from mussel beds. Field surveys within and around three mussel beds showed a peak in cockle densities at 50-100 m toward the coast from the mussel bed, while cockle abundances elsewhere in the study area were very low. Field transplantation of cockles showed higher survival of young cockles (2-3 years old) and increased spat fall coastward of the mussel bed compared to within the bed and to areas without mussels, whereas growth decreased within and coastward of the mussel bed. Our measurements suggest that the observed spatial patterns in cockle numbers resulted from (1) inhibition effects by the mussels close to the beds due to preemptive algal depletion and deteriorated sediment conditions and (2) facilitation effects by the mussels farther away from the beds due to reduction of wave energy. Our results imply that these spatial, scale-dependent interactions between reef-forming ecosystem engineers and surrounding communities of sedentary benthic organisms can be an important determinant of the large-scale community structure in intertidal ecosystems. Understanding this interplay between neighboring communities of sedentary species is therefore essential for effective conservation and restoration of soft-bottom intertidal communities.

Altered membrane lipid composition and functional parameters of circulating cells in cockles (Cerastoderma edule) affected by disseminated neoplasia.

Membrane lipid composition and morpho-functional parameters were investigated in circulating cells of the edible cockle (Cerastoderma edule) affected by disseminated neoplasia (neoplastic cells) and compared to those from healthy cockles (hemocytes). Membrane sterol levels, phospholipid (PL) class and subclass proportions and their respective fatty acid (FA) compositions were determined. Morpho-functional parameters were evaluated through total hemocyte count (THC), mortality rate, phagocytosis ability and reactive oxygen species (ROS) production. Both morpho-functional parameters and lipid composition were profoundly affected in neoplastic cells. These dedifferentiated cells displayed higher THC (5×), mortality rate (3×) and ROS production with addition of carbonyl cyanide m-chloro phenylhydrazone (1.7×) but lower phagocytosis ability (½×), than unaffected hemocytes. Total PL amounts were higher in neoplastic cells than in hemocytes (12.3 and 5.1 nmol×10(-6) cells, respectively). However, sterols and a particular subclass of PL (plasmalogens; 1-alkenyl-2-acyl PL) were present in similar amounts in both cell type membranes. This led to a two times lower proportion of these membrane lipid constituents in neoplastic cells when compared to hemocytes (20.5% vs. 42.1% of sterols in total membrane lipids and 21.7% vs. 44.2% of plasmalogens among total PL, respectively). Proportions of non-methylene interrupted FA- and 20:1n-11-plasmalogen molecular species were the most impacted in neoplastic cells when compared to hemocytes (â…“× and »×, respectively). These changes in response to this leukemia-like disease in bivalves highlight the specific imbalance of plasmalogens and sterols in neoplastic cells, in comparison to the greater stability of other membrane lipid components.

Physiological responses of the European cockle Cerastoderma edule (Bivalvia: Cardidae) as indicators of coastal lagoon pollution.

Physiological responses can be used as effective parameters to identify environmentally stressful conditions. In this study, physiology changes such as oxygen consumption, clearance rate, survival in air, condition index and energy reserves were measured on natural populations of cockles collected from different sites at Ria de Aveiro, Portugal. At those sites, sediment samples were collected for Hg concentration analysis. Cockles were used for the evaluation of both the Hg concentration and physiological response. Mercury was detected in the cockle tissue and in the sediment collected from the sampling points both nearby and distant from the main mercury contamination source. The energy content was negatively correlated with both Hg concentration in cockle tissues and survival in air. Nonetheless, the energy content was positively correlated with the condition index, and there was a positive correlation between the survival in air test and the tissue mercury concentration. A PCA-factor analysis explained 86.8% of the total variance. The principal factor (62.7%) consisted of the air survival, the Hg in soft tissues (positive) and the condition index (negative). The second factor (24.1%) consisted of a negative correlation between the oxygen consumption and the clearance rate. Due to their sensitivity to environmental conditions, the physiological responses of cockles can be used to assess the ecological status of aquatic environments. More effort should be invested in investigating the effects of environmental perturbations on cockle health once they are a good reporter organism.

Impact of digenean parasite infection on metallothionein synthesis by the cockle (Cerastoderma edule): a multivariate field monitoring.

Metallothioneins (MT) are proteins that play an important role in metabolism of essential metals and detoxification of trace metals from living organisms. Their synthesis is induced by metal pollution but can also be exacerbated by other factors such as reproduction processes. In this context, we monitored MT concentrations in a cockle Cerastoderma edule (marine bivalve) population and highlighted the effect of a castrating digenean parasite, Labratrema minimus. In spent cockles, MT levels were low (ca. 5 nmol sites g(-1), fresh weight) but slightly higher in parasitized individuals. During gametogenesis, MT synthesis increased in all cockles, but concentrations were lower in parasitized individuals (18 against 27 nmol sites g(-1), fw in unparasitized cockles) in relation with gonad damage by parasites. Therefore, it is suggested that parasite infection in cockles can modulate MT synthesis that could consequently interfere with the response of these protective proteins in case of metal contamination.