Energy metabolism of juvenile scallops Nodipecten subnodosus under acute increased temperature and low oxygen availability.

High temperature increases energy demand in ectotherms, limiting their physiological capability to cope with hypoxic events. The present study aimed to assess the metabolic tolerance of juvenile Nodipecten subnodosus scallops to acute hyperthermia combined with moderate hypoxia. A previous study showed that juveniles exhibited a high upper temperature limit (32 °C), but the responses of juveniles to combined hyperthermia and low dissolved oxygen are unknown. Scallops were exposed to control conditions (treatment C: 22 °C, ∼7.1 mg O2 L-1 or PO2 156.9 mmHg), acute hyperthermia under normoxia (treatment T: 30 °C, ∼6.0 mg O2 L-1 or PO2 150.9 mmHg) or acute hyperthermia plus hypoxia (treatment TH: 30 °C, ∼2.5 mg O2 L-1 or PO2 62.5 mmHg) for 18 h. In T, juveniles exhibited an enhanced oxygen consumption, together with a decrease in adenylate energy charge (AEC) and arginine phosphate (ArgP), and with no changes in metabolic enzyme activity in the muscle. In TH, scallops maintained similar AEC and ArgP levels in muscle as those observed in T treatment. This response occurred along with the accumulation of inosine monophosphate and hypoxanthine. Besides, reduced citrate synthase and pyruvate kinase activities, enhanced hexokinase activity, and a higher octopine dehydrogenase/lactate dehydrogenase ratio in the mantle indicated the onset of anaerobiosis in TH. These responses indicate that juvenile scallops showed tissue-specific compensatory responses regarding their energy balance under moderate hypoxia at high temperatures. Our results give an insight into the tolerance limit of this species to combined hyperthermia and hypoxia in its northern limit of distribution.

Deciphering the molecular adaptation of the king scallop (Pecten maximus) to heat stress using transcriptomics and proteomics.

BACKGROUND: The capacity of marine species to survive chronic heat stress underpins their ability to survive warming oceans as a result of climate change. In this study RNA-Seq and 2-DE proteomics were employed to decipher the molecular response of the sub-tidal bivalve Pecten maximus, to elevated temperatures. RESULTS: Individuals were maintained at three different temperatures (15, 21 and 25 °C) for 56 days, representing control conditions, maximum environmental temperature and extreme warming, with individuals sampled at seven time points. The scallops thrived at 21 °C, but suffered a reduction in condition at 25 °C. RNA-Seq analyses produced 26,064 assembled contigs, of which 531 were differentially expressed, with putative annotation assigned to 177 transcripts. The proteomic approach identified 24 differentially expressed proteins, with nine identified by mass spectrometry. Network analysis of these results indicated a pivotal role for GAPDH and AP-1 signalling pathways. Data also suggested a remodelling of the cell structure, as revealed by the differential expression of genes involved in the cytoskeleton and cell membrane and a reduction in DNA repair. They also indicated the diversion of energetic metabolism towards the mobilization of lipid energy reserves to fuel the increased metabolic rate at the higher temperature. CONCLUSIONS: This work provides preliminary insights into the response of P. maximus to chronic heat stress and provides a basis for future studies examining the tipping points and energetic trade-offs of scallop culture in warming oceans.

Respiratory response to combined heat and hypoxia in the marine bivalves Pecten maximus and Mytilus spp.

Coastal ecosystems are increasingly disturbed by the increase of mean sea surface temperature and expansion of hypoxic areas. The objectives of the present work were to describe and compare the respiratory responses to combined heat and hypoxia in two bivalve species (Pecten maximus and Mytilus spp.) living in two contrasted coastal habitats (subtidal and intertidal, respectively). Results were consistent with the vertical zonation of both species. Mytilus spp. seemed to cope better with a temperature increase than P. maximus, which was found to be outside of its optimal thermal window at 25°C. Concerning respiratory responses to hypoxia at a given temperature, P. maximus displayed greater oxyregulation capacity that was maintained over a larger range of O2 levels, as compared to Mytilus spp. When acclimation temperatures increased, both species showed a decrease in their oxyregulation capacities alongside a reduction in aerobic performance, especially in P. maximus. The comparison between species suggests that subtidal species, such as P. maximus, might be more vulnerable to a combination of heat and hypoxia than intertidal species, such as Mytilus spp. Lastly, this study highlighted the utility of segmented linear models to estimate PcO2 and regulation percentages in marine organisms exposed to hypoxia.

Effect of low pH on growth and shell mechanical properties of the Peruvian scallop Argopecten purpuratus (Lamarck, 1819).

Dissolution of anthropogenic CO2 modifies seawater pH, leading to ocean acidification, which might affect calcifying organisms such as bivalve mollusks. Along the Peruvian coast, however, natural conditions of low pH (7.6-8.0) are encountered in the habitat of the Peruvian scallop (Argopecten purpuratus), as a consequence of the nearby coastal upwelling influence. To understand the effects of low pH in a species adapted to these environmental conditions, an experiment was performed to test its consequences on growth, calcification, dissolution, and shell mechanical properties in juvenile Peruvian scallops. During 28 days, scallops (initial mean height = 14 mm) were exposed to two contrasted pH conditions: a control with unmanipulated seawater presenting pH conditions similar to those found in situ (pHT = 7.8) and a treatment, in which CO2 was injected to reduce pH to 7.4. At the end of the experiment, shell height and weight, and growth and calcification rates were reduced about 6%, 20%, 9%, and 10% respectively in the low pH treatment. Mechanical properties, such as microhardness were positively affected in the low pH condition and crushing force did not show differences between pH treatments. Final soft tissue weights were not significantly affected by low pH. This study provides evidence of low pH change shell properties increasing the shell microhardness in Peruvian scallops, which implies protective functions. However, the mechanisms behind this response need to be studied in a global change context.

Physiological and comparative proteomic analyzes reveal immune defense response of the king scallop Pecten maximus in presence of paralytic shellfish toxin (PST) from Alexandrium minutum.

The king scallop, Pecten maximus is a highly valuable seafood in Europe. Over the last few years, its culture has been threatened by toxic microalgae during harmful algal blooms, inducing public health concerns. Indeed, phycotoxins accumulated in bivalves can be harmful for human, especially paralytic shellfish toxins (PST) synthesized by the microalgae Alexandrium minutum. Deleterious effects of these toxic algae on bivalves have also been reported. However, its impact on bivalves such as king scallop is far from being completely understood. This study combined ecophysiological and proteomic analyzes to investigate the early response of juvenile king scallops to a short term exposure to PST producing A. minutum. Our data showed that all along the 2-days exposure to A. minutum, king scallops exhibited transient lower filtration and respiration rates and accumulated PST. Significant inter-individual variability of toxin accumulation potential was observed among individuals. Furthermore, we found that ingestion of toxic algae, correlated to toxin accumulation was driven by two factors: 1/ the time it takes king scallop to recover from filtration inhibition and starts to filtrate again, 2/ the filtration level to which king scallop starts again to filtrate after inhibition. Furthermore, at the end of the 2-day exposure to A. minutum, proteomic analyzes revealed an increase of the killer cell lectin-like receptor B1, involved in adaptative immune response. Proteins involved in detoxification and in metabolism were found in lower amount in A. minutum exposed king scallops. Proteomic data also showed differential accumulation in several structure proteins such as ß-actin, paramyosin and filamin A, suggesting a remodeling of the mantle tissue when king scallops are subjected to an A. minutum exposure.

Sources of paralytic shellfish toxin accumulation variability in the Pacific oyster Crassostrea gigas.

This study was designed to assess the contribution of feeding behavior to inter-individual variability of paralytic shellfish toxin (PST) accumulation in the Pacific oyster Crassostrea gigas. For this purpose 42 oysters were exposed for 2 days to non-toxic algae and then for 2 other days to the PST producer Alexandrium minutum. Individual clearance rate (CR) of oysters was continuously monitored over the 4 days using an ecophysiological measurement system. Comparison of CR values when exposed to toxic and non toxic algae allowed to estimate a clearance rate inhibition index (CRII). Toxin concentration of oysters was quantified at the end of the experiment. These data allowed to estimate the toxin accumulation efficiency (TAE) as the ratio of toxin accumulated on toxin consumed. Changes of clearance rate during the experiment indicated that all individuals stopped feeding immediately after being exposed to A. minutum for at least 7 h. This fast response likely corresponded to a behavioral mechanism of avoidance rather to a toxin-induced response. Individuals also showed high inter-variability in their recovery of filtration after this period. Most of the inter-individual variability (78%) in PST accumulation in C. gigas could be explained by the consumption of A. minutum cells, thus emphasizing the importance of the feeding behavior in accumulation. Based on the toxin concentration in their tissues, oysters were clustered in 3 groups showing contrasted patterns of PST accumulation: the high accumulation group was characterized by high feeding rates both on non-toxic and toxic diet and subsequently a low CRII and high TAE. Inversely, the low accumulation group was characterized by low filtration rates, high CRII and low TAE. Both filtration capacity and sensitivity of oysters to toxins may account for the differences in their accumulation. The contribution of TAE in PST accumulation is discussed and might result from differences in assimilation and detoxification abilities among individuals.

Proteomic responses to hypoxia at different temperatures in the great scallop (Pecten maximus).

Hypoxia and hyperthermia are two connected consequences of the ongoing global change and constitute major threats for coastal marine organisms. In the present study, we used a proteomic approach to characterize the changes induced by hypoxia in the great scallop, Pecten maximus, subjected to three different temperatures (10 °C, 18 °C and 25 °C). We did not observe any significant change induced by hypoxia in animals acclimated at 10 °C. At 18 °C and 25 °C, 16 and 11 protein spots were differentially accumulated between normoxia and hypoxia, respectively. Moreover, biochemical data (octopine dehydrogenase activity and arginine assays) suggest that animals grown at 25 °C switched their metabolism towards anaerobic metabolism when exposed to both normoxia and hypoxia, suggesting that this temperature is out of the scallops' optimal thermal window. The 11 proteins identified with high confidence by mass spectrometry are involved in protein modifications and signaling (e.g., CK2, TBK1), energy metabolism (e.g., ENO3) or cytoskeleton (GSN), giving insights into the thermal-dependent response of scallops to hypoxia.

Towards the determination of Mytilus edulis food preferences using the dynamic energy budget (DEB) theory.

The blue mussel, Mytilus edulis, is a commercially important species, with production based on both fisheries and aquaculture. Dynamic Energy Budget (DEB) models have been extensively applied to study its energetics but such applications require a deep understanding of its nutrition, from filtration to assimilation. Being filter feeders, mussels show multiple responses to temporal fluctuations in their food and environment, raising questions that can be investigated by modeling. To provide a better insight into mussel-environment interactions, an experiment was conducted in one of the main French growing zones (Utah Beach, Normandy). Mussel growth was monitored monthly for 18 months, with a large number of environmental descriptors measured in parallel. Food proxies such as chlorophyll a, particulate organic carbon and phytoplankton were also sampled, in addition to non-nutritious particles. High-frequency physical data recording (e.g., water temperature, immersion duration) completed the habitat description. Measures revealed an increase in dry flesh mass during the first year, followed by a high mass loss, which could not be completely explained by the DEB model using raw external signals. We propose two methods that reconstruct food from shell length and dry flesh mass variations. The former depends on the inversion of the growth equation while the latter is based on iterative simulations. Assemblages of food proxies are then related to reconstructed food input, with a special focus on plankton species. A characteristic contribution is attributed to these sources to estimate nutritional values for mussels. M. edulis shows no preference between most plankton life history traits. Selection is based on the size of the ingested particles, which is modified by the volume and social behavior of plankton species. This finding reveals the importance of diet diversity and both passive and active selections, and confirms the need to adjust DEB models to different populations and sites.

Deep sequencing of the mantle transcriptome of the great scallop Pecten maximus.

RNA-Seq transcriptome data were generated from mantle tissue of the great scallop, Pecten maximus. The consensus data were produced from a time course series of animals subjected to a 56-day thermal challenge at 3 different temperatures. A total of 26,064 contigs were assembled de novo, providing a useful resource for both the aquaculture community and researchers with an interest in mollusc shell production.