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Ongoing inputs, in the form of sediment deposition along with associated dissolved contaminants, have challenged the assessment of cap performance at contaminated sediment sites. To address this issue, thin 2–3 cm layer sand caps amended with activated carbon (AC) were investigated for the remediation of polychlorinated biphenyl (PCB) contaminated marine sediments using 90-day mesocosms. All treatments were challenged with (1) ongoing clean or marker-PCB-spiked sediment inputs and (2) bioturbation. Bioaccumulation in hard clams (filter feeding near the cap-water interface) was evaluated to best understand cap effectiveness, relative to sheepshead minnows (confined to the surface water) and sandworms (which burrowed through the caps). All caps (sand and AC amended sand) provided isolation of native bedded PCBs (i.e., PCBs sourced from the bed), reducing uptake in organisms. Total PCB bioaccumulation in clams indicated that AC addition to the cap provided no benefit with spiked influx, or some benefit (56% reduction) with clean influx. Spiked input PCBs, when added to the depositional input sediment, were consistently detected in clams and passive samplers, with and without AC in the cap. PCB uptake by passive samplers located in the caps did not reflect the performance of the remedy, as defined by clam bioaccumulation. However, PCB uptake by passive samplers in the overlying water reasonably represented clam bioaccumulation results.

Intertidal bivalves are constantly exposed to air due to daily and seasonal tidal cycles. The hard clam Mercenaria mercenaria is an economically important bivalve species and often subjected to air exposure for more than 10 days during long-distance transportation. Hard clam exhibits remarkable tolerance to air exposure. In this study, we performed RNA sequencing on hemocytes of M. mercenaria exposed to air for 0, 1, 5, 10, 20 and 30 days. The overall and dynamic molecular responses of hard clams to air exposure were revealed by different transcriptomic analysis strategies. As a result, most cytochrome P450 1A and 3A, and monocarboxylate transporter family members were up-regulated during air exposure. Additionally, the dominant molecular process in response to 5-d, 10-d, 20-d and 30-d air exposure was refolding of misfolded proteins in endoplasmic reticulum, lysosome-mediated degradation of phospholipids , protein metabolism and reorganization of cytoskeleton , and activation of anti-apoptotic process, respectively. Our results facilitated comprehensive understanding of the tolerance mechanisms of intertidal bivalves to air exposure.

Background P-selectin is a molecule participating in the inflammatory response through mediating cellular adhesion and essential for wound repair. However, studies regarding P-selectin in Bivalvia are rare. This study identified 90 P-selectin genes among nine bivalve genomes and classified them into 4 subfamilies according to phylogenetic analysis. Results Notable P-selectin gene expansion was observed in two Venerida species, Sinonovacula constricta and Mercenaria mercenaria. The synteny analysis revealed that P-selectin gene expansion was mostly caused by tandem duplication. In addition, the expression profiles of P-selectin genes in S. constricta showed that many P-selectins were specifically highly expressed in the gills, and the P-selectin expression patterns changed dramatically under low salt stress and ammonia nitrogen stress. Conclusions The massive expansion of P-selectins may facilitate the tolerance to environmental stresses. This study sheds light on the characterizations and expression profiles of P-selectin genes in Bivalvia and provides an integrated framework for further investigation of the role of P-selectins in the environmental tolerance of bivalves.

Lipases form an important group of relevant enzymes which have applications in various fields including; food, pharmaceutical, detergent, textile and cosmetic industries. Lipases can be produced from diverse sources including microorganisms. This study evaluated the potential of bacteria isolates from fresh-water clam Mercenaria Mercenaria to produce lipolytic enzymes. Ten samples of Clam (Mercenaria Mercenaria) were screened for the presence of lipase producing bacteria using classical culture methods. Eleven bacteria species were obtained, of which six (Actinomyces sp., E. coli, Bacillus sp., Pseudomonas sp., Clostridium sp. and Klebsiella sp.) produced lipases that had lipolytic activity in breaking down olive oil used in media supplementation. The best culture media and conditions for optimal production of lipases was studied and it was shown that supplementation of growth media with 2% dextrose at neutral pH gave the greatest yield of lipases when lipase producing isolates were grown in shake flasks. Measurement of biomass by culture and turbidimetric methods indicates that the highest cell mass was recorded by Pseudomonas sp at 7.8 x 105 CFU/ml, closely followed by Actinomyces sp. and Bacillus sp., at 6.2 x 105 CFU/ml and 5.3 x 105 respectively. The produced lipases were partially purified by precipitating with ammonium sulphate followed by dialysis. The total protein content of produced lipases was evaluated by the Lowry’s method, showing that estimated protein content followed the same trend as cell biomass with the highest recorded by Pseudomonas sp. at 1.53mg/ml, followed by Actinomyces sp. and Bacillus sp. at 1.47mg/ml and 1.32mg/ml respectively. The results obtained in this study shows that isolates obtained from freshwater clam can produce potent lipases which can be employed for industrial, food and other diverse uses

Severe losses in aquacultured and wild hard clam (Mercenaria mercenaria) stocks have been previously reported in the northeastern United States due to a protistan parasite called QPX (Quahog Parasite Unknown). Previous work demonstrated that clam resistance to QPX is under genetic control. This study identifies single nucleotide polymorphism (SNP) associated with clam survivorship from two geographically segregated populations, both deployed in an enzootic site. The analysis contrasted samples collected before and after undergoing QPX-related mortalities and relied on a robust draft clam genome assembly. ~200 genes displayed significant variant enrichment at each sampling point in both populations, including 18 genes shared between both populations. Markers from both populations were identified in genes related to apoptosis pathways, protein-protein interaction, receptors, and signaling. This research begins to identify genetic markers associated with clam resistance to QPX disease, leading the way for the development of resistant clam stocks through marker-assisted selection.

With marine diseases on the rise and increased reliance on molecular tools for disease surveillance, validated pathogen detection capabilities are important for effective management, mitigation, and response to disease outbreaks. At the same time, in an era of continual evolution and advancement of molecular tools for pathogen detection, it is critical to regularly reassess previously established assays to incorporate improvements of common practices and procedures, such as the minimum information for publication of quantitative real-time PCR experiments (MIQE) guidelines. Here, we reassessed, re-optimized, and improved the quantitative PCR (qPCR) assay routinely used for Quahog Parasite Unknown (QPX) disease monitoring. We made 19 significant changes to the qPCR assay, including improvements to PCR amplification efficiency, DNA extraction efficiency, inhibition testing, incorporation of linearized standards for absolute quantification, an inter-plate calibration technique, and improved conversion from copy number to number of cells. These changes made the assay a more effective and efficient tool for disease monitoring and pathogen detection, with an improved linear relationship with histopathology compared to the previous version of the assay. To support the wide adoption of validated qPCR assays for marine pathogens, we provide a simple workflow that can be applied to the development of new assays, re-optimization of old or suboptimal assays, or assay validation after changes to the protocol and a MIQE-compliant checklist that should accompany any published qPCR diagnostic assay to increase experimental transparency and reproducibility amongst laboratories.

Background The hard clam Mercenaria mercenaria is a major marine resource along the Atlantic coasts of North America and has been introduced to other continents for resource restoration or aquaculture activities. Significant mortality events have been reported in the species throughout its native range as a result of diseases (microbial infections, leukemia) and acute environmental stress. In this context, the characterization of the hard clam genome can provide highly needed resources to enable basic (e.g., oncogenesis and cancer transmission, adaptation biology) and applied (clam stock enhancement, genomic selection) sciences. Results Using a combination of long and short-read sequencing technologies, a 1.86 Gb chromosome-level assembly of the clam genome was generated. The assembly was scaffolded into 19 chromosomes, with an N50 of 83 Mb. Genome annotation yielded 34,728 predicted protein-coding genes, markedly more than the few other members of the Venerida sequenced so far, with coding regions representing only 2% of the assembly. Indeed, more than half of the genome is composed of repeated elements, including transposable elements. Major chromosome rearrangements were detected between this assembly and another recent assembly derived from a genetically segregated clam stock. Comparative analysis of the clam genome allowed the identification of a marked diversification in immune-related proteins, particularly extensive tandem duplications and expansions in tumor necrosis factors (TNFs) and C1q domain-containing proteins, some of which were previously shown to play a role in clam interactions with infectious microbes. The study also generated a comparative repertoire highlighting the diversity and, in some instances, the specificity of LTR-retrotransposons elements, particularly Steamer elements in bivalves. Conclusions The diversity of immune molecules in M. mercenaria may allow this species to cope with varying and complex microbial and environmental landscapes. The repertoire of transposable elements identified in this study, particularly Steamer elements, should be a prime target for the investigation of cancer cell development and transmission among bivalve mollusks.

Aims: This research was aimed at assessing the nutritional properties and heavy metal compositions of African giant land snails (Archachatina marginata) and clams (Mercenaria mercenaria) from Ekowe community. Methodology: Mineral and vitamin contents were analyzed using the standard method of Association of Analytical Chemist. While standard wet digestion procedure was adopted in the sample preparation, heavy metals were analyzed using the Atomic Absorption Spectrophotometer technique. Results: Moisture contents were 76±0.63% & 78.4±0.06%, ash content, 1.37±0.01% & 3.45±0.01%, crude protein, 18.62±0.74% & 12.74±0.01% and lipid, 1.34±0.01% & 0.07±0.01% for Archachatina marginata and Mercenaria mercenaria respectively. Mineral contents were in this order: Ca > K > PO4> Mg > Na > Fe >Mn for Archachatina marginata while Mercenaria mercenaria was Ca> K > PO4 > Mg > Fe >Mn> Na. Vitamins profiles in Archachatina marginata were 5.20±0.198%, 0.144±0.004%, 0.05±0.003% and 0.78±0.035% for vitamin A, B1, B2 & E respectively. While Mercenaria mercenaria was 3.93±0.070%, 0.13±0.0025%, 0.075±0.001% & 0.84±0.01% for vitamins A, B1, B2 & E respectively. Heavy metal results in Archachatina marginata was in order of Zn > Cu > Cd > Ni > Cr while for Mercenaria mercenaria was Zn > Cu > Cd > Cr > Ni. Conclusion: These metal values were low and within the WHO permissible limits. The result shows that snail could complement the required micro and macro nutrients and vitamins needed for proper growth and development in human and hence recommended for regular consumption.

Ocean uptake of carbon dioxide (CO2) is causing changes in carbonate chemistry that affect calcification in marine organisms. In coastal areas, this CO2-enriched seawater mixes with waters affected by seasonal degradation of organic material loaded externally from watersheds or produced as a response to nutrient enrichment. As a result, coastal bivalves often experience strong seasonal changes in carbonate chemistry. In some cases, these changes may resemble those experienced by aquacultured bivalves during translocation activities. We mimicked these changes by exposing juvenile hard clams (500 μm, Mercenaria mercenaria) to pCO2 in laboratory upwellers at levels resembling those already reported for northeastern US estuaries (mean upweller pCO2 = 773, 1274, and 1838 μatm) and then transplanting to three grow-out sites along an expected nutrient gradient in Narragansett Bay, RI (154 bags of 100 clams). Prior to the field grow-out, clams exposed to elevated pCO2 exhibited larger shells but lower dry weight per unit volume (dw/V). However, percent increase in dw/V was highest for this group during the 27-day field grow-out, suggesting that individuals with low dw/V after the laboratory treatment accelerated accumulation of dw/V when they were transferred to the bay. Treatments also appeared to affect shell mineral structure and condition of digestive diverticula. Although treatment effects diminished during the field grow-out, clams that were pre-exposed for several weeks to high pCO2 would likely have been temporarily vulnerable to predation or other factors that interact with shell integrity. This would be expected to reduce population recovery from short-term exposures to high pCO2.

14 pages, 3 figures, 2 tables.-- This is Smithsonian Marine Station contribution #1138, In 2011, the Indian River Lagoon, a biodiverse estuary in eastern Florida (USA), experienced an intense microalgal bloom with disastrous ecological consequences. The bloom included a mix of microalgae with unresolved taxonomy and lasted for 7 months with a maximum concentration of 130 μg chlorophyll a L. In 2012, brown tide Aureoumbra lagunensis also bloomed in portions of this estuary, with reoccurrences in 2016 and 2018. To identify and understand the role of grazer pressure (top-down control) on bloom formation, we coupled DNA sequencing with bivalve feeding assays using three microalgae isolated from the 2011 bloom and maintained in culture. Feeding experiments were conducted on widely distributed bivalve species in the lagoon, including eastern oysters (Crassostrea virginica), hooked mussels (Ischadium recurvum), charru mussels (Mytella charruana), green mussels (Perna viridis), Atlantic rangia (Rangia cuneata), and hard clams (Mercenaria mercenaria), which were exposed to 3 × 10 cells mL of five species of microalgae consisting of A. lagunensis and the three species clarified herein, the picocyanobacteria Crocosphaera sp. and ‘Synechococcus’ sp., and the picochlorophyte Picochlorum sp., as well as Nannochloropsis oculata used as a control. To ensure clearance rates were indicative of consumption and assimilation, the microalgae were isotopically (N) labeled prior to feeding experiments. Clearance rates differed among bivalve and microalgal species, but enriched N values in bivalve tissue suggest that algal bloom species were assimilated by the bivalves. These results expand our understanding of the important ecosystem services that healthy, biodiverse filter feeder communities provide, We thank St. Johns River Water Management District (Contract 27799) and the Florida Fish and Wildlife Conservation Commission (FWC Agreement No. 13051) for funding the project, With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Circulating hemocytes in the hemolymph represent the backbone of innate immunity in bivalves. Hemocytes are also found in the extrapallial fluid (EPF), the space delimited between the shell and the mantle, which is the site of shell biomineralization. This study investigated the transcriptome, proteome, and function of EPF and hemolymph in the hard clam

In recent decades, the rate of introduction of non-indigenous macroalgae has increased. While invasive seaweeds often outcompete native species for substrata, their direct effects on marine life are rarely described. Here, we describe 'red water' events caused by the decay of blooms of the invasive red seaweed, Dasysiphonia japonica, in Great South Bay, NY, USA, and the ability of water from such events to induce rapid and significant mortality in larval and juvenile fish (Menidia beryllina, Menidia menidia, and Cyprinodon variegatus) and larval bivalves (Mercenaria mercenaria and Crassostrea virginica). All species studied experienced significant (p0.05) reductions in survival when exposed to macroalgae in a state of decay, seawater in which the alga was previously decayed, or both. Both bivalve species experienced 50-60% increases in mortality when exposed to decaying D. japonica for ∼ one week, despite normoxic conditions. Among fish, significant increases (40-80%) in mortality were observed after 24 h exposure to decayed D. japonica and one-week exposures caused, on average, 90% mortality in larval M. beryllina, 50% mortality in juvenile (∼3 cm) M. menidia, and 50% mortality in larval C. variegatus. All fish and bivalve mortality occurred under normoxic conditions (dissolved oxygen (DO)7 mg L

Ruditapes philippinarum and Mercenaria mercenaria are economically important bivalve species. The complete ribosomal DNA (rDNA) unit sequences of R. philippinarum and M. mercenaria, with as-sembled rDNA unit lengths of 12,910 and 12,100 bp, respectively, were obtained in this study for the first time. The rDNA unit structural organisation was similar to that in other eukaryotes, in-cluding the following elements in order: 18S rRNA-internal transcribed spacer 1 (ITS1), 5.8S rRNA-ITS2-28S rRNA-intergenic spacer (IGS) (3′ external transcribed spacer (ETS), non-transcribed spacer (NTS)-5′ ETS). The genetic differences between R. philippinarum and M. mercenaria were mainly attributable to non-coding regions (ITS1, ITS2 and IGS), especially the IGS region. The boundaries of putative 3′ ETS, NTS and 5′ ETS were confirmed. Seven and three sub-repeat fragments were found in R. philippinarum and M. mercenaria, respectively. These frag-ments ranged from 4 to 154 bp in length, and were located at the NTS and 5′ ETS regions. Five and six cytosine–guanine (CpG) islands were detected in R. philippinarum and M. mercenaria, respec-tively, and these covered 85.58% and 79.29% of the entire IGS sequence, respectively. The phylo-genetic tree was constructed based on Veneridae ITS and 18S rRNA sequences using the maxi-mum likelihood (ML) method. The ML tree based on ITS revealed that species within the same genus clearly clustered together with relatively high supporting values, and all the genera were recovered as monophyletic. The phylogenetic analyses using 18S rRNA provided a weaker phy-logenetic signal than ITS.

Oxygen fluctuations are common in marine waters, and hypoxia–reoxygenation (H–R) stress can negatively affect mitochondrial metabolism. The long-lived ocean quahog, Arctica islandica, is known for its hypoxia tolerance associated with metabolic rate depression, yet the mechanisms that sustain mitochondrial function during oxygen fluctuations are not well understood. We used top-down metabolic control analysis (MCA) to determine aerobic capacity and control over oxygen flux in the mitochondria of quahogs exposed to short-term hypoxia (24 h

Biological activity of tissues and enzyme hydrolyzates is investigated for three species of bivalve mollusks:Corbicula japonica,Mercenaria mercenaria, andAnadara broughtonii. Antiradical activity of all samples has increased signifcantly during their enzymatic hydrolysis: in eight times forC. japonica, in three times forM. mercenaria, and in 2.5 times forA. broughtonii. The highest anti-trombolitic activity (0.0823 units) is found for the hydrolyzate ofC. japonicain concentration of 10 mg/mL. Angiotensin I-converting enzyme inhibitory activity is found only for the hydrolyzate ofM. mercenariain concentration of 10–100 mg/mL. The maximum inhibitory activity for alpha-amylase and alpha-glycosidase (7.5 and 7.2 %, respectively) is observed for the hydrolyzate ofM. mercenariain concentration 1 mg/mL. Hepatoprotective activity, measured by survival rate of HepG2 hepatocytes, is detected for the hydrolyzate of tissues for all species in weak concentrations: 0.1 mg/mL forC. japonicaandM. mercenariaand 0.2 mg/mL forA. broughtonii.

Increased consumption and improper disposal of prescription medication, such as beta (β)-blockers, contribute to their introduction into waterways and may pose threats to non-target aquatic organisms. There has been rising concern about the impacts of these prescription drugs on coastal ecosystems, especially because wastewater treatment plants are not designed to eliminate them from the discharge. Few studies have characterized the sublethal effects of β-blocker exposures in marine invertebrates. The overall aim of our research is to identify cellular responses of two commercially important filter-feeding marine bivalves, hard clams (Mercenaria mercenaria) and Eastern oysters (Crassostrea virginica), upon exposures to two β-blocker drugs, propranolol and metoprolol. In vitro exposures with bivalve digestive gland and gill tissues were conducted where tissues were separately exposed to each drug for 24 h. Tissue samples were analyzed for cellular damage (lysosomal membrane destabilization and lipid peroxidation), total antioxidant capacity, and glutathione-s-transferase activity. Elevated damage and changes in enzyme activities were noted in the exposed tissues at environmentally relevant concentrations. Differences in species and tissue sensitivities and responses to exposures were also observed. These studies enhance our understanding of the potential impacts of prescription medication on coastal organisms. Additionally, this work demonstrates that filter-feeders may serve as good model organisms to examine the effects of unintended environmental exposures to β-blockers. These studies are part of our ongoing work aimed at evaluation of sublethal biomarkers of pharmaceutical exposures and identification of key events that can contribute to the development of adverse outcome pathways (AOPs).

Background Inhibitors of apoptosis (IAPs) are critical regulators of programmed cell death that are essential for development, oncogenesis, and immune and stress responses. However, available knowledge regarding IAP is largely biased toward humans and model species, while the distribution, function, and evolutionary novelties of this gene family remain poorly understood in many taxa, including Mollusca, the second most speciose phylum of Metazoa. Results Here, we present a chromosome-level genome assembly of an economically significant bivalve, the hard clam Mercenaria mercenaria, which reveals an unexpected and dramatic expansion of the IAP gene family to 159 members, the largest IAP gene repertoire observed in any metazoan. Comparative genome analysis reveals that this massive expansion is characteristic of bivalves more generally. Reconstruction of the evolutionary history of molluscan IAP genes indicates that most originated in early metazoans and greatly expanded in Bivalvia through both lineage-specific tandem duplication and retroposition, with 37.1% of hard clam IAPs located on a single chromosome. The expanded IAPs have been subjected to frequent domain shuffling, which has in turn shaped their architectural diversity. Further, we observed that extant IAPs exhibit dynamic and orchestrated expression patterns among tissues and in response to different environmental stressors. Conclusions Our results suggest that sophisticated regulation of apoptosis enabled by the massive expansion and diversification of IAPs has been crucial for the evolutionary success of hard clam and other molluscan lineages, allowing them to cope with local environmental stresses. This study broadens our understanding of IAP proteins and expression diversity and provides novel resources for studying molluscan biology and IAP function and evolution.

A study on the distribution of quahog (Mercenaria mercenaria (Linnaeus 1758)) larvae in Narragansett Bay, Rhode Island was conducted during a single summer season in 1995. Samples of the larvae were collected weekly using an electric bilge pump and a 60 ~m mesh plankton net at two depths (0.3 m and 1.6 m) at five sampling stations distributed landward along the West shore, 12.6 km to 30.6 km from Rhode Island Sound. Three stations were located in the upper estuary, i.e., Upper Bay, and two stations were in the lower estuary, i.e., Upper

Color plays a vital function in camouflage, sexual selection, immunity, and evolution. Mollusca possess vivid shell colors and pigmentation starts at the juvenile stage. The hard clam Mercenaria mercenaria is a widely cultivated bivalve of high economic value. To explore the molecular mechanism of pigmentation in juvenile clams, here, we performed RNA-Seq analysis on non-pigmented, white, and red M. mercenaria specimens. Clean reads were assembled into 358,285 transcripts and 149,234 unigenes, whose N50 lengths were 2107 bp and 1567 bp, respectively. Differentially expressed genes were identified and analyzed for KEGG enrichment. “Melanoma/Melanogenesis”, “ABC transporters”, and “Porphyrin and chlorophyll metabolism” pathways appeared to be associated with pigmentation. Pathways related to carotenoid metabolism seemed to also play a vital role in pigmentation in juveniles. Our results provide new insights into the formation of shell color in juvenile hard clams.

Blooms of the dinoflagellate, Cochlodinium (a.k.a.Margalefidinium) polykrikoides, have had deleterious effects on marine life across the Northern Hemisphere and, since the early 1990s, have become more frequent and widespread. While the toxic effects of C. polykrikoides have been well-described for finfish, the effects on bivalve molluscs are poorly understood, particularly in ecosystem and aquaculture settings. The purpose of this study was to characterize the comparative effects of C. polykrikoides blooms on North Atlantic bivalves and to identify the environmental factors that influence its toxic effects. The growth and survival of two age-classes (first- and second-year) of the northern quahog (Mercenaria mercenaria), the bay scallop (Argopecten irradians), and the eastern oyster (Crassostrea virginica) were quantified in surface deployments and at depth during annual bloom events in multiple locations across eastern Long Island (NY, USA), capturing a natural gradient in C. polykrikoides. In two consecutive years, scallops deployed within surface locations experienced significant mortality (75–100%) during short-term (1–2 weeks) but intense (> 1.5 × 104 cells mL−1) C. polykrikoides blooms. Conversely, scallops deployed at depth and clams and oysters deployed at either the surface or at depth were more resistant to blooms. First-year oysters and scallops that survived blooms displayed significant reductions in growth rates, while clams and older scallops and oysters did not. Results suggest that blooms of C. polykrikoides pose significant age- and species-specific threats to native and cultured bivalve shellfish and that shellfish deployed in surface waters are at greater risk during blooms than those deployed at depth.

Providing robust links between mitochondrial genotype and phenotype is of major importance given that mitochondrial DNA (mtDNA) variants can affect reproductive success. Because of the strict maternal inheritance (SMI) of mitochondria in animals, haplotypes that negatively affect male fertility can become fixed in populations. This phenomenon is known as ‘mother's curse’. Doubly uniparental inheritance (DUI) of mitochondria is a stable exception in bivalves, which entails two mtDNA lineages that evolve independently and are transmitted separately through oocytes and sperm. This makes the DUI mitochondrial lineages subject to different sex-specific selective sieves during mtDNA evolution, thus DUI is a unique model to evaluate how direct selection on sperm mitochondria could contribute to male reproductive fitness. In this study, we tested the impact of mtDNA variants on sperm performance and bioenergetics in DUI and SMI species. Analyses also involved measures of sperm performance following inhibition of main energy pathways and sperm response to oocyte presence. Compared to SMI, DUI sperm exhibited (i) low speed and linearity, (ii) a strict OXPHOS-dependent strategy of energy production, and (iii) a partial metabolic shift towards fermentation following egg detection. Discussion embraces the adaptive value of mtDNA variation and suggests a link between male-energetic adaptation, fertilization success and paternal mitochondria preservation.This article is part of the theme issue ‘Linking the mitochondrial genotype to phenotype: a complex endeavour’.

The hard clam (Mercenaria mercenaria) is among the most economically-important marine species along the east coast of the United States, representing the first marine resource in several Northeastern states. The species is rather resilient to infections and the only important disease of hard clams results from an infection caused by Quahog Parasite Unknown (QPX), a protistan parasite that can lead to significant mortality events in wild and aquacultured clam stocks. Though the presence of QPX disease has been documented since the 1960s, little information is available on cellular and molecular interactions between the parasite and the host. This study examined the interactions between the clam immune system and QPX cells. First, the effect of clam plasma on the binding of hemocytes to parasite cells was evaluated. Second, clam plasma proteins that bind QPX cells were identified through proteomic (LC-MS/MS) analyses. Finally, the effect of prior clam exposure to QPX on the abundance of QPX-reactive proteins in the plasma was evaluated. Results showed that plasma factors enhance the attachment of hemocytes to QPX. Among the proteins that specifically bind to QPX cells, several lectins were identified, as well as complement component proteins and proteolytic enzymes. Furthermore, results showed that some of these lectins and complement-related proteins are inducible as their abundance significantly increased following QPX challenge. These results shed light on plasma proteins involved in the recognition and binding of parasite cells and provide molecular targets for future investigations of factors involved in clam resistance to the disease, and ultimately for the selection of resistant clam stocks.

Hypercapnia (elevated CO2 levels) and pollution with trace metals such as Cu and Cd are common stressors in estuarine habitats that can negatively affect physiology and health of marine organisms. Hypercapnia can modulate toxicity of trace metals including Cu and Cd; however, the physiological and cellular mechanisms of the metal-CO2 interactions are not well understood. We investigated the effects of elevated PCO2 (∼800 and 2000μatm) and metal exposure (50μgl-1 of Cu or Cd) on subcellular distribution of metals in two common species of marine bivalves, Eastern oysters Crassostrea virginica and hard shell clams Mercenaria mercenaria. Oysters accumulated higher burdens of Cu and Cd in the gill tissues compared to clams. In both studied species, Cu was predominantly associated with the metabolically active cell compartments (mitochondria, lysosomes, microsomes and cytosolic enzymes), with a modest fraction sequestered by metallothioneins (∼30%) and the insoluble metal-containing granules (MCG) (∼15-20%). Unlike Cu, Cd was largely sequestered by metallothioneins (∼60-70%), with a relatively small fraction associated with the organelles and the cytosolic enzymes. Mitochondria were the main intracellular target for trace metals accumulating higher concentrations of Cd (and in the case of oysters - of Cu) than other organelles or cytosolic enzymes. Cu accumulation in the metabolically active cellular compartments was independent of the CO2 levels, while Cd content of the organelles and cytosolic enzymes increased at elevated PCO2 in both studied species indicating that hypercapnia may enhance cellular toxicity of Cd in bivalves. Hypercapnia suppressed the sequestration capacity of metallothioneins for Cu and Cd in oysters but increased Cu and Cd load in clam metallothioneins. Thus, metal-induced metabolic injury in oysters may be exaggerated by hypercapnia which enhances metal accumulation in the potentially sensitive intracellular fractions and suppresses the metal detoxification capacity. In contrast, clams appear to be more resistant to the combined effects of hypercapnia and metal exposure reflecting more efficient and robust detoxification mechanisms of this species.

Bricelj, V.M.; Kraeuter, J.N., and Flimlin, G., 2017. Status and trends of hard clam, Mercenaria mercenaria, populations in a coastal lagoon ecosystem, Barnegat Bay–Little Egg Harbor, New Jersey. In: Buchanan, G.A.; Belton, T.J., and Paudel, B. (eds.), A Comprehensive Assessment of Barnegat Bay–Little Egg Harbor, New Jersey. This review examines the historical and current status of hard clam, Mercenaria mercenaria, populations in the Barnegat Bay–Little Egg Harbor (BB-LEH) Estuary using New Jersey State stock assessments and published studies and evaluates their potential for rehabilitation under present environmental conditions. This estuary has experienced increasing urbanization, population growth, bulkheading, and changes in watershed use. Clam populations have decreased markedly since the 1960s, and in LEH, substantial areas are now devoid of clams. Landings of wild-caught hard clams, and commercial and recreational clam licenses have all declined. There is no evidence that eutrophication an...

While early life-stage marine bivalves are vulnerable to ocean acidification, effects over successive generations are poorly characterized. The objective of this work was to assess the transgenerational effects of ocean acidification on two species of North Atlantic bivalve shellfish, Mercenaria mercenaria and Argopecten irradians. Adults of both species were subjected to high and low pCO2 conditions during gametogenesis. Resultant larvae were exposed to low and ambient pH conditions in addition to multiple, additional stressors including thermal stress, food-limitation, and exposure to a harmful alga. There were no indications of transgenerational acclimation to ocean acidification during experiments. Offspring of elevated pCO2-treatment adults were significantly more vulnerable to acidification as well as the additional stressors. Our results suggest that clams and scallops are unlikely to acclimate to ocean acidification over short time scales and that as coastal oceans continue to acidify, negative effects on these populations may become compounded and more severe.

As nitrogen entering coastal waters continues to be an issue, much attention has been generated to identify potential options that may help alleviate this stressor to estuaries, including the propagation of bivalves to remove excess nitrogen. Oysters (Crassostrea virginica) and quahogs (Mercenaria mercenaria) from numerous Cape Cod, MA, (USA) sources were analyzed for nitrogen content stored in tissues that would represent a net removal of nitrogen from a water body if harvested. Results showed local oysters average 0.69% nitrogen by total dry weight (mean 0.28gN/animal) and quahogs average 0.67% nitrogen by total dry weight (mean 0.22gN/animal); however, these values did vary by season and to a lesser extent by location or grow-out method. The differences in nitrogen content were largely related to the mass of shell or soft tissue. Nitrogen isotope data indicate shellfish from certain water bodies in the region are incorporating significant amounts of nitrogen from anthropogenic sources.