Soft-shell Clam Mya Arenaria Research Articles

A tale of two soft-shell clams: an integrative taxonomic analysis confirms Mya japonica as a valid species distinct from Mya arenaria (Bivalvia: Myidae)

The soft-shell clam Mya arenaria Linnaeus, 1758 is a commercially important fishery resource that occurs in boreal and temperate environments in the Northern Hemisphere. Whether the soft-shell clam is a single species with a circumboreal range or a species complex also comprising Mya japonica Jay, 1857 distributed in the north Pacific has long been debated by malacologists and palaeontologists based on slight differences in shell morphology. We used an integrative taxonomic approach incorporating available Mya spp. mitochondrial COI and 16S rRNA, and nuclear 28S rRNA gene sequences, as well as spermatozoan and shell morphological characters to test the validity of M. japonica and examine the range of soft-shell clam distribution. Although differences in shell morphology were minor, the results from tree topologies, pairwise uncorrected p-distances, Automatic Barcode Gap Discovery (ABGD) and spermatozoan ultramorphological data confirm the validity of M. japonica in both its endemic region in the northwest Pacific, and as here newly reported introduced populations in British Columbia in the northeast Pacific, and show that M. arenaria is distributed in the northeast Pacific, North Atlantic, Barents Sea (Arctic Ocean) and Mediterranean. We estimate these two closely related sister species diverged 4.1-12.5 Myr during early Pliocene to late Miocene, which is consistent with current evolutionary theory regarding M. arenaria. In addition, ABGD indicated the congener Mya truncata Linnaeus, 1758 may represent a species complex, but additional evidence is still needed to clarify its taxonomic status.

Long-term changes in cohort structure of the soft-shell clam Mya arenaria in the White Sea: growth rate affects lifespan and mortality

ABSTRACTThe population dynamics of a Mya arenaria cohort in the White Sea was studied throughout almost its entire duration (ca. 25 years). We aimed to study changes in the growth heterogeneity of the cohort in terms of individual growth rate and to find out whether there was a connection between the growth rate and both lifespan and mortality rate. Significant variation of the individual growth rate within the cohort of the soft-shell clams was detected. To a great extent it seemed to be determined by the characteristics of the growth achieved in the initial growth period (shell size by the start of the second growing season). Most of the cohort individuals (about 60%) had a similar growth rate during the study period. They belonged to the group of medium-growing clams and with a few exceptions represented the cohort at later stages of development (at the age of 16–24 years). In the most slow-growing group no individuals older than 15 years were found. Clams of this group had the highest mortality rate and the shortest lifespan. In all other groups, periods of low mortality alternated with periods of a much higher mortality. The survival of Mya arenaria seems to be affected mostly by the degree of protection (largely determined by burial depth), intraspecific relationships and ageing.

People of the Clam: Shellfish and Diet in Coastal Maine Late Archaic and Ceramic Period Sites

Abstract Relatively few shell midden sites around the Gulf of Maine have been excavated and analyzed for the quantity of shellfish incorporated into the site. Such data would help us understand the intensity of past shellfish-harvesting pressure on nearby shellfish beds, and the ef fects of shellfish collection on settlement patterns. Moreover , the relative amounts of protein contributed to diet by shellfish versus vertebrates, based on the remains discarded in the midden, indicate that shellfish may have provided the majority of dietary protein. In particular, the softshell clam (Mya arenaria) was of primary importance to regional coastal subsistence and not just a bad-w eather, last-resort food.

Acidification alters predator-prey interactions of blue crab Callinectes sapidus and soft-shell clam Mya arenaria

Acidification due to anthropogenic CO2 pollution, along with episodic or persistent acidification that occurs in coastal environments, will likely result in severe seasonal acidification in estuarine environments. Acidification decreases the fitness of individual species, but the degree to which predator-prey interactions will be impacted is largely unknown. This mesocosm study examined the effect of CO2 acidification on crab-bivalve predator-prey interactions involving two commercially important Chesapeake Bay species, the blue crab Callinectes sapidus and the soft-shell clam Mya arenaria. In particular, the direct effects of CO2 acidification on clam growth and behavior, and the indirect effects of CO2 acidification on interactions between crabs and clams were examined. Mya arenaria were grown in CO2-acidified water (pH7.2) or ambient conditions (pH7.8) for 30days. To determine the effect of acidification on clam responsiveness to mechanical disturbance, a probe was slowly moved towards clams until they ceased pumping (a behavior to avoid detection by predators), and the distance between the probe and the clam's siphon was noted. Clams were exposed to predation by C. sapidus, which were held under acidified or ambient conditions for 48h. Callinectes sapidus handling time, search time, and encounter rate were measured from video. Acidified clams had lighter shells than ambient clams, indicating that shell dissolution occurred. Acidification reduced the responsiveness of M. arenaria to a mechanical disturbance that simulated an approaching predator. As compared to ambient trials, crabs in acidified trials had higher encounter rates; however, this was offset by crabs taking longer to find the first clam in trials, and by increased occurrence of crabs eating only a portion of the prey available. As a result, there was no net change in predation-related clam mortality in acidified trials as compared to ambient conditions. Understanding how acidification will impact food webs in productive estuarine environments requires an examination of the direct impacts of acidification on organism behavior and physiology, as well as indirect effects of acidification mediated through predator-prey interactions.

Large-scale mitochondrial COI gene sequence variability reflects the complex colonization history of the invasive soft-shell clam, Mya arenaria (L.) (Bivalvia)

The aim of the study was to determine genetic diversity in the soft-shell clam Mya arenaria on a wide geographical scale using mtDNA COI gene sequences. Low levels of genetic diversity was found, which can most likely be explained by a bottleneck effect during Pleistocene glaciations and/or selection. The geographical genetic structuring of the studied populations was also very low. The star-like phylogeny of the haplotypes indicates a relatively recent, rapid population expansion following the glaciation period and repeated expansion following the founder effect(s) after the initial introduction of the soft-shell clam to Europe. North American populations are characterized by the largest number of haplotypes, including rare ones, as expected for native populations. Because of the founder effect connected with initial and repeated expansion events, European populations have significantly lower numbers of haplotypes in comparison with those of North America. We also observed subtle differentiations among populations from the North and Baltic seas. The recently founded soft-shell clam population in the Black Sea exhibited the highest genetic similarity to Baltic populations, which confirmed the hypothesis that M. arenaria was introduced to the Gulf of Odessa from the Baltic Sea. The most enigmatic results were obtained for populations from the White Sea, which were characterized by high genetic affinity with American populations.

Location is everything: evaluating the effects of terrestrial and marine resource subsidies on an estuarine bivalve.

Estuaries are amongst the world’s most productive ecosystems, lying at the intersection between terrestrial and marine environments. They receive substantial inputs from adjacent landscapes but the importance of resource subsidies is not well understood. Here, we test hypotheses for the effects of both terrestrial- and salmon-derived resource subsidies on the diet (inferred from stable isotopes of muscle tissue), size and percent nitrogen of the soft-shell clam (Mya arenaria), a sedentary estuarine consumer. We examine how these relationships shift across natural gradients among 14 estuaries that vary in upstream watershed size and salmon density on the central coast of British Columbia, Canada. We also test how assimilation and response to subsidies vary at smaller spatial scales within estuaries. The depletion and enrichment of stable isotope ratios in soft-shell clam muscle tissue correlated with increasing upstream watershed size and salmon density, respectively. The effects of terrestrial- and salmon-derived subsidies were also strongest at locations near stream outlets. When we controlled for age of individual clams, there were larger individuals with higher percent nitrogen content in estuaries below larger watersheds, though this effect was limited to the depositional zones below river mouths. Pink salmon exhibited a stronger effect on isotope ratios of clams than chum salmon, which could reflect increased habitat overlap as spawning pink salmon concentrate in lower stream reaches, closer to intertidal clam beds. However, there were smaller clams in estuaries that had higher upstream pink salmon densities, possibly due to differences in habitat requirements. Our study highlights the importance of upstream resource subsidies to this bivalve species, but that individual responses to subsidies can vary at smaller scales within estuaries.

Comparative study of ecophysiological and biochemical variation between the Baltic and North Sea populations of the invasive soft shell clam Mya arenaria (L. 1758)

Abstract Seasonal variations of environmental factors, such as temperature and salinity, require metabolic acclimatization in sedentary benthic fauna distributed over a wide geographical range. The soft-shell clam Mya arenaria inhabits the coastal waters of the North Atlantic including North America and Europe. In Europe, M. arenaria populations are distributed from Iceland to the Mediterranean Sea, including the North Sea, the Baltic Sea and the Black Sea. Seasonal changes in physiological parameters (gonad index, condition index, biochemical composition and respiration rate) of M. arenaria from the Baltic Sea (the Gulf of Gdańsk, Poland), and the North Sea (Versee Meer, the Netherlands) were studied. The sex ratio of both populations did not differ from 1:1 and the seasonal gonad index was higher in the Baltic population. The average condition index changed seasonally at both studied sites, and was also higher in the Baltic population (except the autumn) compared to the North Sea. In both studied populations, the content of proteins, lipids and carbohydrates in the soft tissue followed the seasonal variations, and it was higher in the Baltic population. The respiration rate was lower in the Baltic population, and seasonal changes in the respiration rate seem to be correlated with changes in the water temperature. Based on the results obtained in the present study, we suggest that Mya arenaria is characterized by a large phenotypic plasticity and differences in the observed physiological traits are due to acclimatization to ambient environmental conditions.

Predation of Sea Scallops and Other Indigenous Bivalves by Invasive Green Crab,Carcinus maenas, from Newfoundland, Canada

ABSTRACT Populations of green crab (Carcinus maenas) have expanded within Newfoundland, and this has raised concern from fish harvesters and scientists regarding bivalve predation in coastal areas on species such as juvenile sea scallops (Placopecten magellanicus). We used 2 microcosm experiments to determine (1) the effects of water temperature (5°C and 12°C) on scallop predation; (2) scallop size selection in small and large green crabs and a large indigenous predator, the rock crab (Cancer irroratus); and (3) bivalve prey selection in large green crabs between softshell clams (Mya arenaria), blue mussels (Mytilus edulis), and sea scallops. Overall, green and rock crabs captured 4 times more scallops in warm water (12°C) than cold (5°C). Large green (60–70 mm) and rock (75–90 mm) crabs captured similar numbers of scallops, selected medium-size (30–40 mm) scallops, and avoided small (10–20 mm) scallops. Small green crabs (40–50 mm) captured only small scallops. Large green crabs selected softshell clams ...

Disseminated neoplasia in the soft-shell clam Mya arenaria: membrane lipid composition and functional parameters of circulating cells.

In a previous study we compared lipid composition and functional parameters of circulating cells from Cerastoderma edule affected or not by disseminated neoplasia (neoplastic cells vs hemocytes) (Le Grand et al. Chem Phys Lipids 167:9-20 2013). Neoplastic cells presented morpho-functional modifications concomitant to striking membrane lipid alterations: the proportion of particular plasmalogen molecular species was drastically decreased. We wanted to test whether this pattern was representative of bivalve neoplastic cells. For the purpose, a similar study was conducted on another bivalve species affected by disseminated neoplasia, the soft-shell clam (Mya arenaria). Although total reactive oxygen species production was unaffected, M. arenaria neoplastic cells presented some functional alterations: phagocytosis activity was reduced by 33%. However, lipid compositions were not drastically altered. Particularly, sterol and plasmalogen levels did not differ between both cell types (about 43% of membrane lipids and 35% of phospholipids, respectively in hemocytes and neoplastic cells). This could be related to the fact that disseminated neoplasia was not related to hemolymph cell proliferation in M. arenaria (0.9±0.2 10(6)cellmL(-1), considering both healthy and neoplastic clams, n=6). Nevertheless this study highlighted minor but specific alterations of membrane lipid composition in M. arenaria neoplastic cells. The only phospholipid subclass in which the fatty acid profile strongly differed between both cell types was serine plasmalogen (PlsSer), with neoplastic cells presenting lower specific enrichment of 20:1n-11 in PlsSer. Such specific alteration of membrane lipid composition strengthened the assumption of an implication of key plasmalogen molecular species in this leukemia-like disease in bivalves.

Patterns of the growth of soft-shell clam Mya arenaria L. (Bivalvia) in shallow water estuaries of the southern Baltic Sea

We explored the shell shape and growth performance of the soft-shell clam Mya arenaria L. in two shallow brackish water estuaries of the southern Baltic Sea – Darss-Zingst and Ruegen lagoons. Observed clams ranged from 2 to 50.1mm in length with detected age up to 6 years old. The growth patterns of the clams were analyzed using the von Bertalanffy growth function (VBGF). An isometric relationship was found between shell length and shell height or shell thickness. Asymptotic length and weight were predicted according to von Bertalanffy equation as L∞=82.75mm and W∞=10.9g. The relationship of wet weight to length was allometric, with exponent b=3.09. Observed shell lengths were close to values predicted by the model. Our study distinguished patterns of the molluscan growth in estuarine transitional environment of the southern Baltic Sea in comparison with other coastal habitats with pronounced spatial environmental heterogeneity.

Distribution, stocks, and age structure of the invader Mya arenaria before and after mass mortalities in the Black Sea

The distribution, stocks, and age structure of the invader soft-shell clam Mya arenaria populations before and after the mass mortalities in the northwestern Black Sea have been studied from 1967 to 1987. At the prolonged influence of mortalities from 1973 to 1985, the number of age classes decreased from six in the period before the mortalities to three in 1985. The restoration of the stocks of Mya arenaria and the possibility of fishing these mollusks after the improvement of the ecological state in this part of the sea are predicted.

Strong population differentiation of softshell clams (Mya arenaria) sampled across seven biogeographic marine ecoregions: possible selection and isolation by distance

Twenty-two Mya arenaria samples spanning seven marine ecoregions mostly situated in the Cold Temperate Northwest Atlantic (CTNA) biogeographic province were collected between 2001 and 2010 and genotyped at seven highly polymorphic microsatellite loci to test for population differentiation. Results showed strong regional differentiation with six genetic clusters: (1) Northern Gulf of St. Lawrence (GSL), (2) Magdalen Archipelago, (3) Southern GSL, (4) Lower Atlantic Canada, (5) US Coasts and (6) Northern Europe. Population structure was supported no matter the statistical approach and generally does not reflect the geographical limits of marine ecoregions. A latitudinal cline in allelic richness provides evidence for a northward post-glacial expansion range for this species. While geographical distance explains the genetic variation detected in southern CTNA, increased heterogeneity observed in northern CTNA can be explained by isolation by distance, marine landscaping and presumable selective processes acting at the Mar5 locus. Exclusion of Mar5 from analyses resulted in the detection of three genetic clusters instead of six.

Transcriptome analysis of neoplastic hemocytes in soft-shell clams Mya arenaria: Focus on cell cycle molecular mechanism

In North America, a high mortality of soft-shell clams Mya arenaria was found to be related to the disease known as disseminated neoplasia (DN). Disseminated neoplasia is commonly recognized as a tetraploid disorder related to a disruption of the cell cycle. However, the molecular mechanisms by which hemocytes of clams are transformed in the course of DN remain by far unknown. This study aims at identifying the transcripts related to DN in soft shell clams’ hemocytes using next generation of sequencing (Illumina HiSeq2000). This study mainly focuses on transcripts and molecular mechanisms involved in cell cycle. Using Illumina next generation of sequencing, more than 95,399,159 reads count with an average length of 45 bp was generated from three groups of hemocytes: (1) a healthy group with less than 10% of tetraploid cells; (2) an intermediate group with tetraploid hemocytes ranging between 10% and 50% and (3) a diseased group with more than 50% of tetraploid cells. After the reads were cleaned by removing the adapters, de novo assembly was performed on the sequences and more than 73,696 contigs were generated with a mean contig length estimated at 585 bp ranging from 189 bp to 14,773 bp. Once a Blastx search against NCBI Non Redundant database was performed and the duplicates removed, 18,378 annotated sequences matched known sequences, 3078 were hypothetical and 9002 were uncharacterized sequences. Fifty percent and 41% of known sequences match sequences from Mollusca and Gastropoda respectively. Among the bivalvia, 33%, 17%, 17% and 15% of the contigs match sequences from Ostreoida, Veneroida, Pectinoida and Mytiloida respectively. Gene ontology analysis showed that metabolic, cellular, transport, cell communication and cell cycle represent 33%, 15%, 9%, 8.5% and 7% respectively of the total biological process. Approximately 70% of the component process is related to intracellular process and 15% is linked to protein and ribonucleoprotein complex. Catalytic activities and binding molecular processes represent 39% and 33% of the total molecular functions. Interestingly, nucleic acid binding represents more than 18% of the total protein class. Transcripts involved in the molecular mechanisms of cell cycle are discussed providing new avenues for future investigations.

Contaminant exposure and ecotoxicological impacts in estuaries

Estuaries are highly dynamic geohydrological systems that form a transition zone between continental rivers and marine environments. They are subjected to many influences such as tides, waves, and influx/efflux of saline water and freshwater. The dynamic mixing of these types of waters provides nutrients in surface waters and sediments making estuaries among the most productive habitats of coastlines. Organisms dwelling in these environments are thus subjected to many combined stresses (salinities, tides and temperature fluctuations) and are particularly at risks to the influence of anthropogenic activity leading to pollution, climate changes and habitats perturbations. Hence, estuaries are area of choice to study the cumulative impacts of pollution in transition zones. A “sustainable partnership” was set up by a France– Quebec initiative in 2003 with the goal to promote and disseminate studies on the wide range of biological responses from organisms exposed to pollution gradients in estuaries of the Atlantic Ocean. This network combines stakeholders involved in both research, teaching, conservation, and protection activities pertaining to science of estuarine ecotoxicology. The success of this endeavor resided on the comparative examination of the impacts of pollution in estuaries from both the Canadian and European coastlines using similar species some cases and focusing on common objectives on either side of the Atlantic in France and Canada (Quebec). The EXECO International Consortium (contaminant exposure and ecotoxicological effects along continental–coastal continuums: http://wwz.ifremer.fr/pollution/Collaborations/GDR-IEXECO ) launched in 2009 has consolidated this important undertaking by combining the actions of two strategic networks: the Canadian (Quebec) CIRE network (Inter-Institutional Ecotoxicology Research Center: http://www.ecotox.uquebec.ca/programmes/htm) and French IMOPHYS network (Integration of Molecular and Physiological Responses to Chemical Contaminants in the Coastal Environment). This special edition on the impacts of pollution in estuarine organisms features a series of articles on work conducted on either side of the Atlantic in the common purpose to gain a better understanding on ecotoxicological effects of anthropogenic activities along the estuarine continuum. These studies featured a blend of both laboratory and field studies at different levels of biological organization and trophic levels. The studies on regulation and adaptation mechanisms were conducted on a molecular basis, e.g., DNA and immune defense alterations, the alteration of physiological functions such as reproduction, and the epidemiological study of chemical contaminant-related tumors. Many papers are based on fish (Platichthys flesus, Microgadus tomcod, Scophtalmus maximus, Limanda limanda, Alosa alosa, Liza ramada), which possess a strong metabolic capacity but the reader will also find papers on bivalve mollusks: the Mytilus edulis mussel and soft shell clam Mya arenaria. Responsible editor: Philippe Garrigues

Selective initiation and transmission of disseminated neoplasia in the soft shell clam Mya arenaria dependent on natural disease prevalence and animal size

Disseminated neoplasia, a diffuse tumor of the hemolymph system, is one of the six most destructive diseases among bivalve mollusk populations, characterized by the development of abnormal, rounded blood cells that actively proliferate. Though the specific etiology of disseminated neoplasia in Mya arenaria remains undetermined, the involvement of viral pathogens and/or environmental pollutants has been suggested and considered. The current study used 5-bromodeoxyuridine (BrDU) known to induce the murine leukemia virus and filtered neoplastic hemolymph to initiate disseminated neoplasia in clams from different populations and size classes respectively. M. arenaria from three locations of different natural neoplasia occurrences were divided into a control and three experimental treatments and injected with 200μl of sterile filtered seawater or 50–200μg/ml BrDU respectively. In a concurrent experiment, animals from different size classes were injected with 2.5% total blood volume of 0.2μm filtered blood from a fully neoplastic animal. Animals were biopsied weekly and cell neoplasia development was counted and scored as 0–25, 26–50, 51–75 and 76–100% neoplastic hemocytes (stages 1–4) in 50μl samples. BrDU injection demonstrated that neoplasia development in M. arenaria was dose dependent on BrDU concentration. In addition, natural disease prevalence at the source location determined initiation of neoplasia induction, with animals from the area of the highest natural disease occurrence displaying fastest neoplasia development (p=0.0037). This could imply that depending on the natural disease occurrence, a potential infectious agent may remain dormant in normal (stage 1) individuals in higher concentrations until activated, i.e. through chemical injection or potentially stress. The size experiment demonstrated that only M. arenaria between 40 and 80mm developed 26–100% neoplastic hemocytes when injected with filtered neoplastic hemolymph, indicating that individuals smaller than 20mm or larger than 80mm were not or no longer susceptible to disease development. So far neoplasia studies have not considered natural disease prevalence or size involvement in neoplasia development and our results indicate that these should be future considerations in neoplasia examinations.

Isolation and Characterization of New Microsatellite Markers for the Invasive Softshell Clam, Mya arenaria (L.) (Bivalvia: Myidae)

The invasive softshell clam (Mya arenaria Linnaeus, 1758) is native to the northwestern region of the Atlantic Ocean. This species has been introduced in the northeast Pacific and along the European coasts, due to intense naval transports and aquaculture, and it is now present in all the European seas. In this paper we describe seven new microsatellite loci for Mya arenaria. The isolated loci are polymorphic with a number of alleles per locus between 6 and 14. The observed and expected heterozygosities ranged from 0.417 to 0.951, and from 0.643 to 0.895, with an average of 0.716 and 0.775, respectively. These microsatellite markers should be useful in analyzing this species’ genetic diversity, which could explain various processes of its invasion history.

Expression of RAS-like family members, c-jun and c-myc mRNA levels in neoplastic hemocytes of soft-shell clams Mya arenaria using microsphere-based 8-plex branched DNA assay

The molecular mechanisms by which disseminated neoplasia (DN) is developed in soft shell clams Mya arenaria remain largely unknown. This study aims at quantifying Rho-like GTPase, RAS-Rho, RAS-related C3 botulinum (RAS C3), c-jun as well as c-myc transcript levels in clams sampled at North River (Charlottetown, Prince Edward Island, Canada). The transcripts were quantified using multiplex gene analysis (Quantigene® 2 Plex, Affymetrix) in 3 groups of clams: (1) Group C (healthy clams considered as control) with a low percentage of tetraploid hemocytes (<10%); (2) Group D (disease in development): individuals presenting a percentage of tetraploid cells ranging between 10% and 50%; (3) Group E (established disease): clams with a high percentage of tetraploid hemocytes (>50%). Data showed a down-regulation of Rho-like GTPase, Rho-like subfamily, RAS C3, c-jun and an up-regulation of c-myc gene expression. It is believed that a deregulation of the expression of these genes could partly unravel the molecular mechanisms involved in the development of DN in soft shell clams Mya arenaria. Further investigations should be pursued to determine the role of these gene products in clams' hemocytes.

Modeling the Effect of Environmental Parameters on Feeding Ecology of the Shortnose Sturgeon in the Saint John River, New Brunswick

AbstractWe examined the effects of spatial, temporal, and environmental parameters on the feeding habits of the shortnose sturgeon Acipenser brevirostrum. We sampled sturgeon gut contents using colonic flushing, a new, low‐impact technique. We found that heavy feeding continued until at least November, when water temperature reached 8°C. In all seasons, foraging shortnose sturgeon used shallow water (2–8 m); however, in late fall this trend was even stronger, when 50% of the sturgeon were captured in less than 3 m depth. Over 60 families of prey items were identified from 166 colonic samples collected at four points between spring and late fall from seven locations throughout the lower Saint John River, New Brunswick. Samples from freshwater sites contained mainly Gastropoda, Sphaeriidae, Chironomidae, and Gammaridae. Samples from saline and brackish environments contained mostly Gammaridae, Corophiidae, softshell clam Mya arenaria, Cyathura polita, and Chironomidae. We used sensitivity analysis to determine the number of shortnose sturgeon required to obtain a representative sample of the diet. We found that 20–40 gut samples from sturgeons in both brackish and freshwater environments would suffice for a general description of the diet but that complete knowledge of the sturgeon diet and a comparison among several sites would require the sampling of hundreds of fish. The digestibility of prey varied from 77% of the ingested volume in a saline environment to only 6% in freshwater samples. Shortnose sturgeon from freshwater sites had higher ingested volumes and more prey per ingested volume than did those from brackish sites, which indicated further differences in feeding efficiency between the environments. Salinity was a significant parameter in the redundancy analysis of ingested prey items. In freshwater sites the significant factors were season, sturgeon length, ingested volume, number of ingested prey, and number of ingested taxa. In saline sites the significant factors were season, location, ingested volume, and the number of ingested prey. Our findings identified the spatial and temporal gradients in shortnose sturgeon foraging and provided insight into foraging efficiency in this species.

Lack of detection of a putative retrovirus associated with haemic neoplasia in the soft shell clam Mya arenaria

Haemic neoplasia (HN) is a leukemia-like disease that affects at least 20 species of marine bivalves including soft shell clam, Mya arenaria. Since the disease was discovered in 1969, the etiology remains unknown. A retroviral etiology has been suggested based on the detection of reverse transcriptase activity and electron microscopic observation of retroviral-like particles using negative staining. To date, however no virus isolate and no retroviral sequence from HN has been obtained. Moreover, transmission of the disease by cell-free filtrate from affected clams has not been reproduced. In the current study, we reinvestigated the association of HN with a putative retrovirus. Sucrose gradient centrifugation followed by assessment of reverse transcriptase activity, electrophoretic analysis of protein and RNA, and electron microscopic examinations of fractions corresponding to retroviral density were employed. Detection of retroviral pol sequences using degenerate RT-PCR approaches was also attempted. Our results showed visible bands at the expected density of retrovirus in HN-positive and HN-negative clam tissues and both with reverse transcriptase activity. Electron microscopy, RNA analysis, protein analysis, and PCR systems targeting the pol gene of retroviruses did not however provide clear evidence supporting presence of a retrovirus. We point out that the retrovirus etiology of HN of Mya arenaria proposed some 25years ago should be reconsidered in the absence of a virus isolate or virus sequences.

In vitro interactions between several species of harmful algae and haemocytes of bivalve molluscs

Harmful algal blooms (HABs) can have both lethal and sublethal impacts on shellfish. To understand the possible roles of haemocytes in bivalve immune responses to HABs and how the algae are affected by these cells (haemocytes), in vitro tests between cultured harmful algal species and haemocytes of the northern quahog (= hard clam) Mercenaria mercenaria, the soft-shell clam Mya arenaria, the eastern and Pacific oysters Crassostrea virginica and Crassostrea gigas and the Manila clam Ruditapes philippinarum were carried out. Within their respective ranges of distribution, these shellfish species can experience blooms of several HAB species, including Prorocentrum minimum, Heterosigma akashiwo, Alexandrium fundyense, Alexandrium minutum and Karenia spp.; thus, these algal species were chosen for testing. Possible differences in haemocyte variables attributable to harmful algae and also effects of haemolymph and haemocytes on the algae themselves were measured. Using microscopic and flow cytometric observations, changes were measured in haemocytes, including cell morphology, mortality, phagocytosis, adhesion and reactive oxygen species (ROS) production, as well as changes in the physiology and the characteristics of the algal cells, including mortality, size, internal complexity and chlorophyll fluorescence. These experiments suggest different effects of the several species of harmful algae upon bivalve haemocytes. Some harmful algae act as immunostimulants, whereas others are immunosuppressive. P. minimum appears to activate haemocytes, but the other harmful algal species tested seem to cause a suppression of immune functions, generally consisting of decreases in phagocytosis, production of ROS and cell adhesion and besides cause an increase in the percentage of dead haemocytes, which could be attributable to the action of chemical toxins. Microalgal cells exposed to shellfish haemolymph generally showed evidence of algal degradation, e.g. loss of chlorophyll fluorescence and modification of cell shape. Thus, in vitro tests allow a better understanding of the role of the haemocytes and the haemolymph in the defence mechanisms protecting molluscan shellfish from harmful algal cells and could also be further developed to estimate the effects of HABs on bivalve molluscs in vivo.