Mesocosm study of PAC-modified clay effects on Karenia brevis cells and toxins, chemical dynamics, and benthic invertebrate physiology.

Modified clay compounds are used globally as a method of controlling harmful algal blooms, and their use is currently under consideration to control Karenia brevis blooms in Florida, USA. In 1400 L mesocosm tanks, chemical dynamics and lethal and sublethal impacts of MC II, a polyaluminum chloride (PAC)-modified kaolinite clay, were evaluated over 72 h on a benthic community representative of Sarasota Bay, which included blue crab (Callinectes sapidus), sea urchin (Lytechinus variegatus), and hard clam (Mercenaria campechiensis). In this experiment, MC II was dosed at 0.2 g L-1 to treat bloom-level densities of K. brevis at 1 × 106 cells L-1. Cell removal in MC II-treated tanks was 57% after 8 h and 95% after 48 h. In the water column, brevetoxin analogs BTx-1 and BTx-2 were found to be significantly higher in untreated tanks at 24 and 48 h, while in MC II-treated tanks, BTx-3 was found to be higher at 48 h and BTx-B5 was found to be higher at 24 and 48 h. In MC II floc, we found no significant differences in BTx-1 or BTx-2 between treatments for any time point, while BTx-3 was found to be significantly higher in the MC II-treated tanks at 48 and 72 h, and BTx-B5 was higher in MC II-treated tanks at 24 and 72 h. Among various chemical dynamics observed, it was notable that dissolved phosphorus was consistently significantly lower in MC II tanks after 2 h, and that turbidity in MC II tanks returned to control levels 48 h after treatment. Dissolved inorganic carbon and total seawater alkalinity were significantly reduced in MC II tanks, and partial pressure of CO2 (pCO2) was significantly higher in the MC II-only treatment after 2 h. In MC II floc, particulate phosphorus was found to be significantly higher in MC II tanks after 24 h. In animals, lethal and sublethal responses to MC II-treated K. brevis did not differ from untreated K. brevis for either of our three species at any time point, suggesting MC II treatment at this dosage has negligible impacts to these species within 72 h of exposure. These results appear promising in terms of the environmental safety of MC II as a potential bloom control option, and we recommend scaling up MC II experiments to field trials in order to gain deeper understanding of MC II performance and dynamics in natural waters.

The hard clam genome reveals massive expansion and diversification of inhibitors of apoptosis in Bivalvia.

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.

Single-molecule long-read (SMRT) transcriptome sequencing of Mercenaria mercenaria reveals a powerful anti-apoptotic system critical for air exposure endurance.

Mercenaria mercenaria is an economically important clam species and exhibits an outstanding resistance to multiple environmental stressors. However, our understanding of their stress adaptability is limited due to a lack of genomic information, such as transcriptome resources. In this study, single-molecule long-read (SMRT) mRNA sequencing was performed to obtain the full-length gill transcriptome reference sequences of M. mercenaria under air exposure stress. In all, 14.5 G subreads were obtained and assembled into 64,603 unigenes, among which 50,613 were successfully annotated. Additionally, 56,295 SSRs, 1457 transcription factors, and 5924 lncRNAs were identified in M. mercenaria transcriptome. Furthermore, numerous apoptosis-related transcripts were identified according to Swiss-Prot annotation and their numbers were counted. We also found that most apoptosis-related transcripts exhibited typical domains of a certain protein family through conserved domain prediction. Additionally, eight typical sequences related to apoptosis pathway were detected by RT-PCR, with the aim to show the sequential variation of gene expression levels under air exposure. These results implied that the complicated apoptosis system, especially the powerful anti-apoptotic system was critical for M. mercenaria to endure air exposure.

Effects of seawater salinity and pH on cellular metabolism and enzyme activities in biomineralizing tissues of marine bivalves.

Molluscan shell formation is a complex energy demanding process sensitive to the shifts in seawater CaCO3 saturation due to changes in salinity and pH. We studied the effects of salinity and pH on energy demand and enzyme activities of biomineralizing cells of the Pacific oyster (Crassostrea gigas) and the hard-shell clam (Mercenaria mercenaria). Adult animals were exposed for 14 days to high (30), intermediate (18), or low (10) salinity at either high (8.0-8.2) or low (7.8) pH. Basal metabolic cost as well as the energy cost of the biomineralization-related cellular processes were determined in isolated mantle edge cells and hemocytes. The total metabolic rates were similar in the hemocytes of the two studied species, but considerably higher in the mantle cells of C. gigas compared with those of M. mercenaria. Cellular respiration was unaffected by salinity in the clams' cells, while in oysters' cells the highest respiration rate was observed at intermediate salinity (18). In both studied species, low pH suppressed cellular respiration. Low pH led to an upregulation of Na+/K+ ATPase activity in biomineralizing cells of oysters and clams. Activities of Ca2+ ATPase and H+ ATPase, as well as the cellular energy costs of Ca2+ and H+ transport in the biomineralizing cells were insensitive to the variation in salinity and pH in the two studied species. Variability in cellular response to low salinity and pH indicates that the disturbance of shell formation under these conditions has different underlying mechanisms in the two studied species.

Potential nitrous oxide production by marine shellfish in response to warming and nutrient enrichment.

Bivalves facilitate microbial nitrogen cycling, which can produce nitrous oxide (N2O), a potent greenhouse gas. Potential N2O production by three marine bivalves (Mytilus edulis, Mercenaria mercenaria and Crassostrea virginica) was measured in the laboratory including responses to nitrogen (N) loading and/or warming over short-terms (up to 14 or 28 days). N additions (targeting 100 µM-N ammonium nitrate) or warming (22 °C) individually and in combination were applied with experimental controls (20 µM-N, 19 °C). N2O production rates were higher with N additions for all species, but warming lacked significant direct effects. Ammonium and nitrate concentrations varied but were consistent with nitrification as a potential N2O source for all bivalves. Highest N2O emissions (7.5 nmol N2O g-1 h-1) were from M. edulis under hypoxic conditions coincident with a drop in pH. Macro-epifauna on M. edulis did not significantly alter N2O production. Thus, under short-term hypoxic conditions, micro-organisms in M. edulis guts may be a particularly significant source of N2O.

Modeling the odor-landscape resulting from the pumping behavior of bivalve clams in the presence of predators.

Motivated by experimental findings, a computational fluid dynamics (CFD) model was used to investigate whether the clam Mercenaria mercenaria may alter its cue downstream variability by an exhalant random pumping behavior. This behavior was hypothesized to occur in the presence of predator chemical signals in order to prevent successful tracking by the predator. Simulated downstream flow and mixing conditions derived from the random nature of the clam exhalant jet in a crossflow were analyzed by computing an intermittency factor, determining the field of finite-time Lyapunov exponents (FTLEs) and identifying the resulting Lagrangian coherent structures (LCSs). Numerical simulations illustrate that the effectiveness of a fluctuating exhalant jet to prevent downstream tracking by a crab, depends on the ratio of the exhalant jet to the crossflow. Specifically, the clam could effectively enhance the downstream dispersion to prevent tracking, but only in the range of parameters where LCSs are generated (jet-to-crossflow ratio ≥ 1). Then, the probability of detection is reduced with respect to the case of a less fluctuating exhalant jet.

Stimpson's hard clam Mercenaria stimpsoni; A multi-decadal climate recorder for the northwest Pacific coast.

A sclerochronological and radiocarbon-based study of life history traits of Stimpson's hard clam (Mercenaria stimpsoni), collected alive from Funakoshi Bay, northeast Japan, showed the lifespan of the species to be at least 92 years (determined from annual growth line counts). Three M. stimpsoni specimens exhibited the following synchronous growth pattern, suggestive of environmental control; annual increment width increasing after 1955 to a maximum value between 1970 and 1980, subsequently decreasing gradually until 2000, and thereafter remaining constant or increasing slightly. Variations on annual growth patterns, as well as standardized growth indices chronology, were relatively closely linked to the Atlantic Multidecadal Oscillation (AMO), but less so to Pacific Decadal Oscillation (PDO). Carbonate samples collected from ontogenetically younger shell portions, estimated from growth line counts to have been deposited before 1950, contained no nuclear bomb-test radiocarbon, thereby supporting the accuracy of annual growth line counts (versus overcounting from ventral margin). Together with the synchronous annual increment width patterns, this indicated that age and annual growth rate estimations for M. stimpsoni based on growth line counts were reliable and applicable to high-resolution sclerochronological analyses, which should contribute to a deeper understanding of multi-decadal northwest Pacific climate variability.

Hard clam walking: Active horizontal locomotion of adult Mercenaria mercenaria at the sediment surface and behavioral suppression after extensive sampling.

Locomotion of infaunal bivalve mollusks primarily consists of vertical movements related to burrowing; horizontal movements have only been reported for a few species. Here, we characterize hard clam walking: active horizontal locomotion of adults (up to 118 mm shell length, SL) of the commercially important species, Mercenaria mercenaria, at the sediment surface-a behavior only briefly noted in the literature. We opportunistically observed walking over a 10-yr period, at 9 different sites in the Peconic Bays, New York, USA, and tested several hypotheses for the underlying cause of this behavior through quantitative field sampling and reproductive analyses. Hard clam walking was exhibited by males and females at equal frequency, predominantly during June/July and October, when clams were in peak spawning condition. Extensive walking behavior appears to be cued by a minimum population density; we suggest it may be mediated by unidentified pheromone(s), infaunal pressure waves and/or other unidentified factors. There was no directionality exhibited by walking clams, but individuals in an area of extensive walking were highly aggregated and walking clams were significantly more likely to move toward a member of the opposite sex. Thus, we conclude that hard clam walking serves to aggregate mature individuals prior to spawning, thereby facilitating greater fertilization success. In the process of investigating this behavior, however, we apparently oversampled one population and reduced clam densities below the estimated minimum threshold density and, in so doing, suppressed extensive walking for a period of >3 years running. This not only reinforces the importance of detailed field investigations of species biology and ecology, even for those that are considered to be well studied, but also highlights the need for greater awareness of the potential for research activities to affect focal species behavior.

Intermittent hypoxia leads to functional reorganization of mitochondria and affects cellular bioenergetics in marine molluscs.

Fluctuations in oxygen (O2) concentrations represent a major challenge to aerobic organisms and can be extremely damaging to their mitochondria. Marine intertidal molluscs are well-adapted to frequent O2 fluctuations, yet it remains unknown how their mitochondrial functions are regulated to sustain energy metabolism and prevent cellular damage during hypoxia and reoxygenation (H/R). We used metabolic control analysis to investigate the mechanisms of mitochondrial responses to H/R stress (18 h at <0.1% O2 followed by 1 h of reoxygenation) using hypoxia-tolerant intertidal clams Mercenaria mercenaria and hypoxia-sensitive subtidal scallops Argopecten irradians as models. We also assessed H/R-induced changes in cellular energy balance, oxidative damage and unfolded protein response to determine the potential links between mitochondrial dysfunction and cellular injury. Mitochondrial responses to H/R in scallops strongly resembled those in other hypoxia-sensitive organisms. Exposure to hypoxia followed by reoxygenation led to a strong decrease in the substrate oxidation (SOX) and phosphorylation (PHOS) capacities as well as partial depolarization of mitochondria of scallops. Elevated mRNA expression of a reactive oxygen species-sensitive enzyme aconitase and Lon protease (responsible for degradation of oxidized mitochondrial proteins) during H/R stress was consistent with elevated levels of oxidative stress in mitochondria of scallops. In hypoxia-tolerant clams, mitochondrial SOX capacity was enhanced during hypoxia and continued rising during the first hour of reoxygenation. In both species, the mitochondrial PHOS capacity was suppressed during hypoxia, likely to prevent ATP wastage by the reverse action of FO,F1-ATPase. The PHOS capacity recovered after 1 h of reoxygenation in clams but not in scallops. Compared with scallops, clams showed a greater suppression of energy-consuming processes (such as protein turnover and ion transport) during hypoxia, indicated by inactivation of the translation initiation factor EIF-2α, suppression of 26S proteasome activity and a dramatic decrease in the activity of Na(+)/K(+)-ATPase. The steady-state levels of adenylates were preserved during H/R exposure and AMP-dependent protein kinase was not activated in either species, indicating that the H/R exposure did not lead to severe energy deficiency. Taken together, our findings suggest that mitochondrial reorganizations sustaining high oxidative phosphorylation flux during recovery, combined with the ability to suppress ATP-demanding cellular functions during hypoxia, may contribute to high resilience of clams to H/R stress and help maintain energy homeostasis during frequent H/R cycles in the intertidal zone.

Soundscapes and Larval Settlement: Larval Bivalve Responses to Habitat-Associated Underwater Sounds.

We quantified the effects of habitat-associated sounds on the settlement response of two species of bivalves with contrasting habitat preferences: (1) Crassostrea virginicia (oyster), which prefers to settle on other oysters, and (2) Mercenaria mercenaria (clam), which settles on unstructured habitats. Oyster larval settlement in the laboratory was significantly higher when exposed to oyster reef sound compared with either off-reef or no-sound treatments. Clam larval settlement did not vary according to sound treatments. Similar to laboratory results, field experiments showed that oyster larval settlement in "larval housings" suspended above oyster reefs was significantly higher compared with off-reef sites.

Effects of contaminated St. Lucie River saltwater sediments on an amphipod (Ampelisca abdita) and a hard-shell clam (Mercenaria mercenaria).

The St. Lucie estuary (SLE) ecosystem in South Florida has been shown to be contaminated with metals and pesticides. Our earlier studies also showed that aquatic organisms, especially benthic species in the SLE ecosystem, might be potentially at high risk from copper (Cu) exposure. The objectives of this study were to conduct studies with separate groups of organisms exposed to seven field-collected sediment samples from the St. Lucie River according to standard procedures to evaluate toxicity and tissue concentrations of Cu and zinc (Zn). Short term and longer term whole sediment acute toxicity studies were performed with Ampelisca abdita and Mercenaria mercenaria. Analysis of sediment chemical characteristics showed that Cu and Zn are of most concern because their concentrations in 86 % of the sediments were higher than the threshold effect concentrations for Florida sediment quality criteria and the National Oceanic and Atmospheric Administration Screening Quick Reference Tables (SQuiRTs) sediment values. There was no significant effect on survival of the tested organisms. However, increased Cu and Zn concentrations in the test organisms were found. Dry weight of the tested organisms was also inversely related to Cu and Zn concentrations in sediments and organisms. The effects on organism weight and Cu and Zn uptake raise concerns about the organism population dynamics of the ecosystem because benthic organisms are primary food sources in the SLE system and are continuously exposed to Cu- and Zn-contaminated sediments throughout their life cycle. The results of the present study also indicate that Cu and Zn exposures by way of sediment ingestion are important routes of exposure.

A non-native prey mediates the effects of a shared predator on an ecosystem service.

Non-native species can alter ecosystem functions performed by native species often by displacing influential native species. However, little is known about how ecosystem functions may be modified by trait-mediated indirect effects of non-native species. Oysters and other reef-associated filter feeders enhance water quality by controlling nutrients and contaminants in many estuarine environments. However, this ecosystem service may be mitigated by predation, competition, or other species interactions, especially when such interactions involve non-native species that share little evolutionary history. We assessed trophic and other interference effects on the critical ecosystem service of water filtration in mesocosm experiments. In single-species trials, typical field densities of oysters (Crassostrea virginica) reduced water-column chlorophyll a more strongly than clams (Mercenaria mercenaria). The non-native filter-feeding reef crab Petrolisthes armatus did not draw down chlorophyll a. In multi-species treatments, oysters and clams combined additively to influence chlorophyll a drawdown. Petrolisthes did not affect net filtration when added to the bivalve-only treatments. Addition of the predatory mud crab Panopeus herbstii did not influence oyster feeding rates, but it did stop chlorophyll a drawdown by clams. However, when Petrolisthes was also added in with the clams, the clams filtered at their previously unadulterated rates, possibly because Petrolisthes drew the focus of predators or habituated the clams to crab stimuli. In sum, oysters were the most influential filter feeder, and neither predators nor competitors interfered with their net effect on water-column chlorophyll. In contrast, clams filtered less, but were more sensitive to predators as well as a facilitative buffering effect of Petrolisthes, illustrating that non-native species can indirectly affect an ecosystem service by aiding the performance of a native species.

Interactive effects of mosquito control insecticide toxicity, hypoxia, and increased carbon dioxide on larval and juvenile eastern oysters and hard clams.

Mosquito control insecticide use in the coastal zone coincides with the habitat and mariculture operations of commercially and ecologically important shellfish species. Few data are available regarding insecticide toxicity to shellfish early life stages, and potential interactions with abiotic stressors, such as low oxygen and increased CO2 (low pH), are less understood. Toxicity was assessed at 4 and 21 days for larval and juvenile stages of the Eastern oyster, Crassostrea virginica, and the hard clam, Mercenaria mercenaria, using two pyrethroids (resmethrin and permethrin), an organophosphate (naled), and a juvenile growth hormone mimic (methoprene). Acute toxicity (4-day LC50) values ranged from 1.59 to >10 mg/L. Overall, clams were more susceptible to mosquito control insecticides than oysters. Naled was the most toxic compound in oyster larvae, whereas resmethrin was the most toxic compound in clam larvae. Mortality for both species generally increased with chronic insecticide exposure (21-day LC50 values ranged from 0.60 to 9.49 mg/L). Insecticide exposure also caused sublethal effects, including decreased swimming activity after 4 days in larval oysters (4-day EC50 values of 0.60 to 2.33 mg/L) and decreased growth (shell area and weight) in juvenile clams and oysters after 21 days (detected at concentrations ranging from 0.625 to 10 mg/L). Hypoxia, hypercapnia, and a combination of hypoxia and hypercapnia caused mortality in larval clams and increased resmethrin toxicity. These data will benefit both shellfish mariculture operations and environmental resource agencies as they manage the use of mosquito control insecticides near coastal ecosystems.

A heart that beats for 500 years: age-related changes in cardiac proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in Arctica islandica, the longest-living noncolonial animal.

Study of negligibly senescent animals may provide clues that lead to better understanding of the cardiac aging process. To elucidate mechanisms of successful cardiac aging, we investigated age-related changes in proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in the heart of the ocean quahog Arctica islandica, the longest-lived noncolonial animal (maximum life span potential: 508 years). We found that in the heart of A. islandica the level of oxidatively damaged proteins did not change significantly up to 120 years of age. No significant aging-induced changes were observed in caspase-like and trypsin-like proteasome activity. Chymotrypsin-like proteasome activity showed a significant early-life decline, then it remained stable for up to 182 years. No significant relationship was observed between the extent of protein ubiquitination and age. In the heart of A. islandica, an early-life decline in expression of HSP90 and five mitochondrial electron transport chain complexes was observed. We found significant age-related increases in the expression of three cytokine-like mediators (interleukin-6, interleukin-1ß, and tumor necrosis factor-α) in the heart of A. islandica. Collectively, in extremely long-lived molluscs, maintenance of protein homeostasis likely contributes to the preservation of cardiac function. Our data also support the concept that low-grade chronic inflammation in the cardiovascular system is a universal feature of the aging process, which is also manifest in invertebrates.

Bioelectric fields of marine organisms: voltage and frequency contributions to detectability by electroreceptive predators.

Behavioral responses of elasmobranch fishes to weak electric fields have been well studied. These studies typically employ a stimulator that produces a dipole electric field intended to simulate the natural electric field of prey items. However, the characteristics of bioelectric fields have not been well described. The magnitude and frequency of the electric field produced by 11 families of marine organisms were quantified in this study. Invertebrate electric potentials ranged from 14 to 28 µV and did not differ from those of elasmobranchs, which ranged from 18 to 30 µV. Invertebrates and elasmobranchs produced electric potentials smaller than those of teleost fishes, which ranged from 39 to 319 µV. All species produced electric fields within the frequency range that is detectable by elasmobranch predators (<16 Hz), with the highest frequencies produced by the penaeids (10.3 Hz) and the gerreids (4.6 Hz). Although voltage differed by family, there was no relationship between voltage and mass or length of prey. Differences in prey voltage may be related to osmoregulatory strategies; invertebrates and elasmobranchs are osmoconformers and have less ion exchange with the surrounding seawater than teleosts species, which are hyposmotic. As predicted, voltage production was greatest at the mucous membrane-lined mouth and gills, which are sites of direct ion exchange with the environment.

Interactive effects of elevated temperature and CO(2) levels on metabolism and oxidative stress in two common marine bivalves (Crassostrea virginica and Mercenaria mercenaria).

Marine bivalves such as the hard shell clams Mercenaria mercenaria and eastern oysters Crassostrea virginica are affected by multiple stressors, including fluctuations in temperature and CO2 levels in estuaries, and these stresses are expected to be exacerbated by ongoing global climate change. Hypercapnia (elevated CO2 levels) and temperature stress can affect survival, growth and development of marine bivalves, but the cellular mechanisms of these effects are not yet fully understood. In this study, we investigated whether oxidative stress is implicated in cellular responses to elevated temperature and CO2 levels in marine bivalves. We measured the whole-organism standard metabolic rate (SMR), total antioxidant capacity (TAOC), and levels of oxidative stress biomarkers in the muscle tissues of clams and oysters exposed to different temperatures (22 and 27°C) and CO2 levels (the present day conditions of ~400ppm CO2 and 800ppm CO2 predicted by a consensus business-as-usual IPCC emission scenario for the year 2100). SMR was significantly higher and the antioxidant capacity was lower in oysters than in clams. Aerobic metabolism was largely temperature-independent in these two species in the studied temperature range (22-27°C). However, the combined exposure to elevated temperature and hypercapnia led to elevated SMR in clams indicating elevated costs of basal maintenance. No persistent oxidative stress signal (measured by the levels of protein carbonyls, and protein conjugates with malondialdehyde and 4-hydroxynonenal) was observed during the long-term exposure to moderate warming (+5°C) and hypercapnia (~800ppm CO2). This indicates that long-term exposure to moderately elevated CO2 and temperature minimally affects the cellular redox status in these bivalve species and that the earlier observed negative physiological effects of elevated CO2 and temperature must be explained by other cellular mechanisms.

Extreme longevity is associated with increased resistance to oxidative stress in Arctica islandica, the longest-living non-colonial animal.

We assess whether reactive oxygen species production and resistance to oxidative stress might be causally involved in the exceptional longevity exhibited by the ocean quahog Arctica islandica. We tested this hypothesis by comparing reactive oxygen species production, resistance to oxidative stress, antioxidant defenses, and protein damage elimination processes in long-lived A islandica with the shorter-lived hard clam, Mercenaria mercenaria. We compared baseline biochemical profiles, age-related changes, and responses to exposure to the oxidative stressor tert-butyl hydroperoxide (TBHP). Our data support the premise that extreme longevity in A islandica is associated with an attenuated cellular reactive oxygen species production. The observation of reduced protein carbonyl concentration in A islandica gill tissue compared with M mercenaria suggests that reduced reactive oxygen species production in long-living bivalves is associated with lower levels of accumulated macromolecular damage, suggesting cellular redox homeostasis may determine life span. Resistance to aging at the organismal level is often reflected in resistance to oxidative stressors at the cellular level. Following TBHP exposure, we observed not only an association between longevity and resistance to oxidative stress-induced mortality but also marked resistance to oxidative stress-induced cell death in the longer-living bivalves. Contrary to some expectations from the oxidative stress hypothesis, we observed that A islandica exhibited neither greater antioxidant capacities nor specific activities than in M mercenaria nor a more pronounced homeostatic antioxidant response following TBHP exposure. The study also failed to provide support for the exceptional longevity of A islandica being associated with enhanced protein recycling. Our findings demonstrate an association between longevity and resistance to oxidative stress-induced cell death in A islandica, consistent with the oxidative stress hypothesis of aging and provide justification for detailed evaluation of pathways involving repair of free radical-mediated macromolecular damage and regulation of apoptosis in the world's longest-living non-colonial animal.

In vivo effects of brown tide on the feeding function of the gill of the northern quahog Mercenaria mercenaria (Bivalvia: Veneridae).

The in vivo response of adult northern quahogs, Mercenaria mercenaria, to Aureococcus anophagefferens (brown tide) at the level of the gill was determined using video-endoscopy. Feeding activity, particle-approach velocities, and ventral-groove-transport velocities were documented after the quahogs were exposed to Isochryis galbana (baseline observations) supplemented with either toxic or nontoxic A. anophagefferens at two bloom concentrations (8 x 10(5) or 2 x 10(6) cells ml(-1)). Externally, there was no evidence of adverse effects of brown tide on feeding, as siphons remained extended and dilated. Toxic brown tide at both concentrations elicited gill muscular contractions, intermittent cessation of water flow, and decreased particle loading within the pallial cavity. The 8 x 10(5) cell ml(-1) toxic treatment had no significant effect on approach velocities or ventral-groove-transport velocities after 2 h, although time-averaging showed significant reduction of the latter during the last 30 min of exposure. The higher concentration of toxic brown tide caused a significant decrease in these velocities after only 1 h. Nontoxic brown tide produced none of these effects. Thus, A. anophagefferens compromised quahog feeding by stimulating contractions of the branchial musculature and interfering with lateral and ventral groove ciliary beating. These effects were both time- and concentration-dependent and could be caused by either a dopaminergic or a serotonergic toxic factor.

Use of the juvenile clam, Mercenaria mercenaria, as a sensitive indicator of aqueous and sediment toxicity.

The hard clam is an economically important bivalve and is abundant along the East Coast of the US. The goal of this research was to evaluate the sensitivity of this test species as compared to that of other benthic and epibenthic organisms. Toxic effects of cadmium (inorganic metal), DDT (organochlorine pesticide), and fluoranthene (polycyclic aromatic hydrocarbon) exposure in sediments (10-day) and seawater (24-h) on juvenile (212-350-microm) hard clams Mercenaria mercenaria were determined. The aqueous 24-h LC(50) values were 0.42 mg/L cadmium (95% CL=0.35-0.45 mg/L), 0.61 mg/L DDT (95% CL=0.40-0.95 mg/L), and 0.65 mg/L fluoranthene (95% CL=0.44-1.23 mg/L). Results of sediment toxicity tests indicated that the 10-day LC(50) values were 1.66 mg/kg cadmium (95% CL=1.21-2.28 mg/kg), 5.8 mg/kg DDT (95% CL=4.8-8.3mg/kg), and 1.75 mg/kg fluoranthene (95% CL=1.38-2.09 mg/kg). Based on comparisons to toxicity data for other marine species, these findings suggest that the juvenile clam is one of the more sensitive species to a variety of contaminants and may be a valuable indicator for potential sediment toxicity.

Toxicity of atrazine to the juvenile hard clam, Mercenaria mercenaria.

The herbicide atrazine is one of the most heavily used pesticides in the United States. The effects of atrazine on the clam Mercenaria mercenaria were evaluated in aqueous and sediment laboratory assays. Juvenile clams of approximately 1mm in size were used for all experiments. An acute aqueous bio-assay was used to determine the 96-h LC(50) for the juvenile clams. A chronic aqueous bioassay was conducted at lower atrazine concentrations over a 10-day exposure period to examine both lethal and sublethal (dry mass, shell size, and condition index) endpoints. A chronic sediment bioassay examined mortality and sublethal endpoints in a 10-day exposure. The acute 96-h LC(50) was 5608 microg/L with 95% confidence intervals ranging from 5003 to 6287 microg/L. Results of the chronic aqueous assay indicated both lethal and sublethal (reduced shell size) effects at high atrazine concentrations. In the 10-day chronic aqueous assay, the no observable effect concentration was 500 microg/L, the lowest observable effect concentration was 1000 microg/L, and the maximum allowable toxicant concentration (MATC) was 707 microg/L. There were no significant effects of atrazine in the chronic sediment exposure. Safe concentrations for the aqueous experiments were estimated by applying an uncertainty factor of 10 to the calculated MATC values. While there were adverse effects of atrazine at high concentrations, these results suggest that atrazine is not directly toxic to M. mercenaria at environmentally relevant concentrations.