Exposure to Alexandrium spp. impairs the development of Green-lipped mussel (Perna canaliculus) embryos and larvae.

The green-lipped mussel (GLM) Perna canaliculus is an economically, ecologically, and culturally important species in Aotearoa New Zealand. Since 2011, harmful algal blooms (HABs) of Alexandrium spp. have occurred annually in the Marlborough Sounds, the largest GLM aquaculture region in New Zealand. Across a similar timeframe, there has been a severe reduction in wild spat (juvenile mussel) catch. This research investigated the effects of Alexandrium pacificum (which produces paralytic shellfish toxins; PSTs) and A. minutum (a non-producer of PSTs) on the development of four GLM larval life stages (gametes, embryos, D-stage and settlement). Early life stages of GLM were exposed to environmentally relevant concentrations of Alexandrium spp. as whole cell, lysate and filtrate treatments. A 48-h exposure of embryos to whole A. pacificum cells at 500 cells mL-1 caused lysis of embryos, severe abnormalities, and reduced development through to veliger (D-stage) larvae by 85%. GLM growth was impaired at cell concentrations as low as 250 cells mL-1 during a 4-day exposure of D-stage larvae to both Alexandrium spp. Exposure of GLM to both whole and lysed treatments of Alexandrium spp. at 500 cells mL-1 resulted in halved larval growth rates (2.00 µm day-1 vs 4.48 µm day-1 in the control) and growth remained impeded during a 4-day recovery period. Both A. pacificum and A. minutum were found to negatively impact D-larvae. Both whole-cell and lysed-cell treatments of A. pacificum had similar negative effects, suggesting that Alexandrium spp. toxicity to D-larvae is independent of PSTs. Additionally, cell membrane-free treatments of A. pacificum had no negative effects on embryo development, indicating that cell surface-associated bioactive compounds may be responsible for the observed negative effects during this early life stage. Conversely, non-PST-producing A. minutum was toxic to D-stage larvae but not to embryos; larval growth was reduced following a brief 1 h exposure of sperm to cell membrane-free treatments of A. pacificum. No effects were recorded in GLM larvae exposed during settlement, highlighting the potential for differences in susceptibility of early life stages to Alexandrium spp. exposure and the influence of exposure durations. In the wild, blooms of Alexandrium spp. can persist for several months, reaching cell densities higher than those investigated in the present study, and as such may be detrimental to the vulnerable early life stages of GLM.

Inactivated ostreid herpesvirus-1 induces an innate immune response in the Pacific oyster, Crassostrea gigas, hemocytes.

Infectious diseases are a major constraint to the expansion of shellfish production worldwide. Pacific oyster mortality syndrome (POMS), a polymicrobial disease triggered by the Ostreid herpesvirus-1 (OsHV-1), has devastated the global Pacific oyster (Crassostrea gigas) aquaculture industry. Recent ground-breaking research revealed that C. gigas possess an immune memory, capable of adaption, which improves the immune response upon a second exposure to a pathogen. This paradigm shift opens the door for developing 'vaccines' to improve shellfish survival during disease outbreaks. In the present study, we developed an in-vitro assay using hemocytes - the main effectors of the C. gigas immune system - collected from juvenile oysters susceptible to OsHV-1. The potency of multiple antigen preparations (e.g., chemically and physically inactivated OsHV-1, viral DNA, and protein extracts) to stimulate an immune response in hemocytes was evaluated using flow cytometry and droplet digital PCR to measure immune-related subcellular functions and gene expression, respectively. The immune response to the different antigens was benchmarked against that of hemocytes treated with Poly (I:C). We identified 10 antigen preparations capable of inducing immune stimulation in hemocytes (ROS production and positively expressed immune- related genes) after 1 h of exposure, without causing cytotoxicity. These findings are significant, as they evidence the potential for priming the innate immunity of oysters using viral antigens, which may enable cost-effective therapeutic treatment to mitigate OsHV-1/POMS. Further testing of these antigen preparations using an in-vivo infection model is essential to validate promising candidate pseudo-vaccines.

Understanding the Dynamic of POMS Infection and the Role of Microbiota Composition in the Survival of Pacific Oysters, Crassostrea gigas.

For over a decade, Pacific oyster mortality syndrome (POMS), a polymicrobial disease, induced recurring episodes of massive mortality affecting Crassostrea gigas oysters worldwide. Recent studies evidenced a combined infection of the ostreid herpesvirus (OsHV-1 µVar) and opportunistic bacteria in affected oysters. However, the role of the oyster microbiota in POMS is not fully understood. While some bacteria can protect hosts from infection, even minor changes to the microbial communities may also facilitate infection and worsen disease severity. Using a laboratory-based experimental infection model, we challenged juveniles from 10 biparental oyster families with previously established contrasted genetically based ability to survive POMS in the field. Combining molecular analyses and 16S rRNA gene sequencing with histopathological observations, we described the temporal kinetics of POMS and characterized the changes in microbiota during infection. By associating the microbiota composition with oyster mortality rate, viral load, and viral gene expression, we were able to identify both potentially harmful and beneficial bacterial amplicon sequence variants (ASVs). We also observed a delay in viral infection resulting in a later onset of mortality in oysters compared to previous observations and a lack of evidence of fatal dysbiosis in infected oysters. Overall, these results provide new insights into how the oyster microbiome may influence POMS disease outcomes and open new perspectives on the use of microbiome composition as a complementary screening tool to determine shellfish health and potentially predict oyster vulnerability to POMS. IMPORTANCE For more than a decade, Pacific oyster mortality syndrome (POMS) has severely impacted the Crassostrea gigas aquaculture industry, at times killing up to 100% of young farmed Pacific oysters, a key commercial species that is cultivated globally. These disease outbreaks have caused major financial losses for the oyster aquaculture industry. Selective breeding has improved disease resistance in oysters, but some levels of mortality persist, and additional knowledge of the disease progression and pathogenicity is needed to develop complementary mitigation strategies. In this holistic study, we identified some potentially harmful and beneficial bacteria that can influence the outcome of the disease. These results will contribute to advance disease management and aquaculture practices by improving our understanding of the mechanisms behind genetic resistance to POMS and assisting in predicting oyster vulnerability to POMS.

Characterization of the effects of triclosan on sperm and embryos of Mytilus and Perna mussel species.

The Greenshell™ mussel (GSM), Perna canaliculus, is a culturally and commercially important species in New Zealand. Declines in spat settlement of GSM have been observed in important growing areas and the cause(s) have not been identified. One hypothesis is that chemical contaminants could be a contributing factor. The aim to this study was to investigate the effects of acute exposure on early life stages using the anti-microbial triclosan (TCS) as a benchmark toxicant and the blue mussel (BM), Mytilus galloprovincialis, as a reference species. Sperm and embryos of BM and GSM were exposed to TCS for 1 h and 48 h, respectively. Following exposures, a range of parameters were investigated including spermatozoa cellular characteristics via flow cytometry, fertilization success, larval mortality and size. Exposure to TCS negatively impacted functional parameters of sperm, reduced the fertilization success and larval size, and increased larval mortality in both BM and GSM with LC5048h of 94.3 and 213 µg L-1, respectively. Triclosan increased sperm ROS production in both species, which could cause destabilisation of mitochondrial and other cellular membranes, resulting in reduced mitochondrial membrane potential (BM) and increased sperm size (GSM), leading to apoptosis in both species. Fertilization success of GSM was only affected at the highest TCS concentration tested (391 µg L-1), but development of larvae derived from exposed sperm was affected from the lowest concentrations tested (0.5 and 5.2 µg L-1) in both species. This highlights the importance of assessing the sensitivity of contaminants across developmental stages. Results of this study confirm that TCS causes oxidative stress and has membranotropic effects, and that early life stages of the endemic GSM are suitable to assess ecotoxicity of contaminants such as TCS.

Dietary Exposure of Pacific Oyster (Crassostrea gigas) Larvae to Compromised Microalgae Results in Impaired Fitness and Microbiome Shift.

The Pacific oyster Crassostrea gigas is the world's most cultivated oyster and seed supply is heavily reliant on hatchery production where recurring mass mortality events are a major constraint. Outbreaks of bacterial infection via microalgal feed are frequently implicated in these mortalities. This study assessed the effects of feeding compromised microalgae to developing oyster larvae. Intentionally 'stressed' (high pH) or non-stressed microalgae were fed to 11 day-old oyster larvae at two feeding rations for 96 h, followed by a recovery period. Biological endpoints of larval performance were measured following the 96 h exposure and subsequent recovery. Bacterial communities associated with the microalgae feed, rearing seawater, and the oyster larvae, were characterized and correlated with effects on oyster fitness parameters. Feeding stressed algae to oyster larvae for 96 h increased the occurrence of deformities (>70% vs. 20% in control), reduced feeding and swimming ability, and slowed development. Following the recovery period, fewer larvae reached pediveliger stage (2.7% vs. 36% in control) and became spat (1.5% vs. 6.6% in control). The quantity of stressed algae supplied to oyster larvae also influenced overall larval performance, with high feeding rations generally causing greater impairment than low rations. Bacterial profiling using 16S rRNA showed that most bacterial families characterized in larval tissue were also present in larval rearing seawater and in the microalgae feed (98%). The rearing seawater showed the highest bacterial richness compared to the larval and the microalgal compartments, regardless of feeding regime. In larval tissue, bacterial richness was highest in stressed and high-feed treatments, and negatively correlated with larval fitness parameters. These results suggest significant dysbiosis induced by compromised feed and/or increased feed ration. Several bacterial genera (e.g., Halomonas, Marinomonas) were strongly associated with impaired larval performance while the presence of genera in larvae including Vibrio was closely associated with overfeeding. Our research demonstrated that metabarcoding can be effectively used to identify microbiota features associated with larval fitness.

Balancing essential and non-essential metal bioavailability during hatchery rearing of Greenshell mussel (Perna canaliculus) larvae.

The use of ethylenediaminetetraacetic acid (EDTA) during bivalve hatchery production is thought to improve larval yields due to the reduced exposure to toxic metals (such as Cu); however, few studies have focused on the bioavailability of metals during the rearing process. Greenshell™ mussels (Perna canaliculus) were reared for 48 h with and without EDTA (12 µM) exposure and larvae were subsequently raised to 21 days post-fertilisation with and without EDTA exposure. Survival, shell length, algal ingestion rate, swimming activity, total metal concentration in water, bioavailable metal concentrations and larval metal accumulation were monitored for the 21 day period. Larval fitness (specifically D-yields) was improved on day 2 in the EDTA treatment, whereas an overall negative effect of EDTA treatment on fitness was observed on day 10 and 21. During the first 48 h, increased survival in the EDTA treatment is believed to be due to the reduction of bioavailable Zn concentrations in the rearing seawater. No other metal (essential or non-essential) displayed a consistent trend when comparing metal bioavailability to any of the fitness parameters measured throughout the experiment. Though the measured metal bioavailability was not clearly linked to fitness, the uptake of Al, P, Cr, Fe, Co, Ni, Zn, As, Cd, and Hg by P. canaliculus was reduced during the first 48 h, suggesting that the biological regulation of these elements is just as important as the bioavailability. Overall, treatment of the rearing seawater with 12 µM EDTA is effective for improving Greenshell™ mussel larval yields by decreasing metal bioavailability during the first two days of development but has minimal benefit between day 2 and 21.

Development of a method to cryopreserve Greenshell mussel™ (Perna canaliculus) veliger larvae.

Cryopreservation of larvae of Greenshell™ mussel Perna canaliculus, the most cultivated species in New Zealand, can provide flexibility for selective breeding programmes and enhance its global production. In this study, we set out to develop a reliable protocol for freezing D-stage larvae of Greenshell™ mussels that ensured long-term survival for successful rearing of thawed larvae in the hatchery. The effects of different combinations of cryoprotecting agents (CPA), varying CPA equilibration times, larval concentrations per straw as well as different larval development stages (48 h vs 72 h old) were evaluated by assessing the behavioural response (swimming activity, algal consumption), shell size and survival of larvae, up to 4 days post-thawing. The protocol yielding the best larval performances was a combination of the following CPA (final concentrations): 14% ethylene-glycol (EG) + 0.6 M trehalose (TRE) + 1% polyvinyl-pyrrolidone (PVP), prepared with Milli-Q water. Stocking densities ranging from 50,000 to 150,000 larvae per straw (0.25 mL) and a 20 min equilibration time gave the best results, while no significant differences in fitness were found between larvae cryopreserved at 48 h nor 72 h-old. Using the improved cryopreservation protocol, over 50% of previously cryopreserved D-larvae were able to survive after 4 days of rearing, compared with 65% in the unfrozen control. More importantly, about one third of thawed larvae were able to swim and feed, and to potentially develop further. These findings contribute to enhance the selective breeding programmes for this species.

Cryopreservation of sea urchin sperm and early life stages.

Sea urchins are excellent model organisms useful for several lines of biotechnological research. Sea urchins are typically collected from the sea and kept in research facilities all over the world for such purposes. Cryopreservation can be a very powerful tool to enhance the use of sea urchins as a model species for research. The development of cryopreservation protocols for different sea urchin gametes, embryos, and larvae allows year round access to high quality material outside the natural reproductive season. It also reduces the uncertainty and variability that may be caused by changing oceanic, meteorological, and environmental conditions. Access to cryopreserved gametes and embryos will allow using these model organisms in laboratories all around the world, regardless of their facilities or their proximity to a natural population of sea urchins. Cryopreservation is a very useful tool for aquaculture production, fisheries conservation, and wild stock enhancement allowing spat supply all year round without the need of conditioning broodstock for out of season reproduction-which is expensive, time consuming, and often unfruitful. It will also provide flexibility for selective breeding programs by allowing crossings of individuals with different reproductive seasons. Although cryopreservation protocols have been successfully developed for many valuable fisheries species such as sturgeons, salmonid fishes, and other marine invertebrates (e.g., oysters), only a small amount of research has been carried out regarding sea urchin cryopreservation. In this chapter, we outline protocols for the cryopreservation of sea urchin cells.

Sublethal effects of oil-contaminated sediment to early life stages of the Eastern oyster, Crassostrea virginica.

The explosion of the Deepwater Horizon (DWH) oil drilling rig resulted in the release of crude oil into the Gulf of Mexico. This event coincided with the spawning season of the Eastern oyster, Crassostrea virginica. Although oil bound to sediments constitutes an important source of polycyclic aromatic hydrocarbon (PAH) exposure to benthic organisms, toxicity of sediment-associated DWH oil has not been investigated in any bivalve species. Here, we evaluated the sublethal effects of acute exposure of gametes, embryos and veliger larvae of the Eastern oyster to different concentrations of unfiltered elutriates of sediment contaminated with DWH oil. Our results suggest that gametes, embryos and veliger larvae are harmed by exposure to unfiltered elutriates of contaminated sediment. Effective concentrations for fertilization inhibition were 40.6 µg tPAH50 L-1 and 173.2 µg tPAH50 L-1 for EC201h and EC501h values, respectively. Embryo exposure resulted in dose-dependent abnormalities (EC20 and EC50 values were 77.7 µg tPAH50 L-1 and 151 µg tPAH50 L-1, respectively) and reduction in shell growth (EC2024h value of 1180 µg tPAH50 L-1). Development and growth of veliger larvae were less sensitive to sediment-associated PAHs compared to embryos. Fertilization success and abnormality of larvae exposed as embryos were the most sensitive endpoints for assessing the toxicity of oil-contaminated sediment. Bulk of measured polycyclic aromatic hydrocarbons were sediment-bound and caused toxic effects at lower tPAH50 concentrations than high energy water accommodated fractions (HEWAF) preparations from the same DWH oil. This study suggests risk assessments would benefit from further study of suspended contaminated sediment.

Impacts of exposure to the toxic dinoflagellate Karenia brevis on reproduction of the northern quahog, Mercenaria mercenaria.

The Gulf of Mexico, including the southwest Florida coast, USA, experience recurrent blooms of the brevetoxin (PbTx)-producing dinoflagellate, Karenia brevis. Northern quahogs (hard clams) Mercenaria mercenaria, are an important commercial species in this region. This study examined the effects of field and laboratory exposure of adult clams to K. brevis during their reproductive period, and effects on their subsequently produced offspring. Ripe adult clams were collected from a site which had been exposed to an eight-month natural bloom of K. brevis and an unaffected reference site. Ripe adult clams were also exposed to bloom concentrations of K. brevis for 10 days in the laboratory. Clams exposed to K. brevis accumulated PbTx at concentrations of 1508 (field exposure), 1444 (1000 cells mL-1 laboratory treatment) and 5229 ng g-1 PbTx-3 eq (5000 cells mL-1 laboratory treatment). Field-exposed clams showed histopathological effects: a significantly higher prevalence of mucus in the stomach/ intestine (23.3%), edema in gill tissues (30%) and presence of the cestode parasite, Tylocephalum spp. in whole tissue (40%), compared to non-exposed clams (0, 3.3 and 6.7% respectively). These clams also showed reduced gonadal allocation (23% gonadal area) and a higher prevalence of clams of undetermined sex (20%) compared to those sampled from the non-exposed site (43% and 0%, respectively). It is hypothesized that less energy may be channeled into reproduction as more is allocated for homeostasis or tissue repair. The fertilization success of gametes obtained from both field and laboratory-exposed adults was significantly lower in clams that had been exposed to K. brevis and development of these offspring was negatively affected at Days 1 and 4 post-fertilization (in field- and laboratory-exposed clams at the higher K. brevis concentration and in laboratory-exposed clams at the higher K. brevis concentration, respectively). Negative effects may be due to toxin accumulation in the gametes of field-exposed clams (244 ± 50 ng PbTx g-1 and 470 ± 82 ng g-1 wet weight in oocytes and sperm, respectively). Adverse effects in M. mercenaria are compared to those previously reported in oysters, Crassostrea virginica, under similar conditions of exposure. This study provides further evidence of the impacts of K. brevis and its associated toxins on the adults and offspring of exposed shellfish. Site-selection for the collection of broodstock and aquaculture grow-out efforts should therefore consider the local occurrence of K. brevis blooms.

Evaluation of toxicity of Deepwater Horizon slick oil on spat of the oyster Crassostrea virginica.

The 2010 explosion of the Deepwater Horizon (DWH) oil rig generated the largest marine oil spill in US history with millions of barrels of crude oil released in the Gulf of Mexico (GoM). The eastern oyster, Crassostrea virginica, is an ecologically and economically important species in the northern GoM. Due to its biological characteristics (sessile, filter feeding), juvenile oysters may have been affected. This study investigated the effects of surface-collected DWH oil prepared as high-energy water-accommodated fraction (HEWAF) on the survival of 2-month-old oyster spat, and evaluated the potential impacts of HEWAF on particle clearance rate and spat tissue. Exposure of oysters to a range of oil/HEWAF (0-7-66-147-908-3450 µg tPAH50 (sum of 50 polycyclic aromatic hydrocarbons) L-1) resulted in non-dose-dependent mortalities and reduced clearance rates of algal food (Tisochrysis lutea). A morphometric study of the digestive tubules (DGTs) indicated a dose-dependent response to oil exposure on lumen dilation, on epithelium thinning of the DGT, and a significant change in DGT synchrony (LOEC = 66 µg tPAH50 L-1). This finding suggests that structural changes occurred in the digestive gland of exposed oysters most likely due to an oil-related stress. In addition, histological observations showed that tissues in contact with HEWAF (gills, palp, connective tissue, digestive gland) were adversely impacted at ≥ 7 µg tPAH50 L-1, and exhibited pathological symptoms typical of an inflammatory response (e.g., hemocyte diapedesis and infiltration, syncytia, epithelium sloughing).

Effects of field and laboratory exposure to the toxic dinoflagellate Karenia brevis on the reproduction of the eastern oyster, Crassostrea virginica, and subsequent development of offspring.

Blooms of the brevetoxin-producing dinoflagellate, Karenia brevis, are a recurrent and sometimes devastating phenomenon in the Gulf of Mexico. The eastern oyster, Crassostrea virginica, is exposed regularly to these blooms, yet little is known about the impacts of K. brevis upon this important species. The present study considered the effects of exposure to both a natural bloom and cultured K. brevis on the reproductive development of C. virginica. Oysters had been exposed to a bloom of K. brevis that occurred in Lee County, Florida, from September 2012 through May 2013, during a period of gametogenesis and gamete ripening. Ripe adult oysters were collected from this bloom-exposed site and from a site 200 miles north which was not exposed to any bloom. In addition, responses to two 10-day laboratory exposures of either unripe or ripe adult oysters to whole cells of K. brevis at high bloom concentrations (1000 and 5000cellsmL-1) were determined. Both field- and laboratory-exposed adult oysters accumulated PbTx (attaining ∼22×103ngg-1 and 922ngg-1 PbTx-3 equivalents in the laboratory and the field, respectively), and significant mucal, edematous, and inflammatory features, indicative of a defense response, were recorded in adult tissues in direct contact with K. brevis cells. Laboratory-exposed oysters also showed an increase in the total number of circulating hemocytes suggesting that: (1) new hemocytes may be moving to sites of tissue inflammation, or, (2) hemocytes are released into the circulatory system from inflamed tissues where they may be produced. The area of oyster tissue occupied by gonad (representative of reproductive effort) and reactive oxygen species production in the spermatozoa of oysters exposed to the natural bloom of K. brevis were significantly lower compared to oysters that were not exposed to K. brevis. Additionally, following 10-day exposure of ripe oysters, a significant, 46% reduction in the prevalence of individuals with ripe gametes was obtained in the 5000cellsmL-1K. brevis treatment. Brevetoxin (PbTx) was recorded within the spermatozoa and oocytes of naturally exposed oysters and was estimated to be 18 and 26% of the adult PbTx load, respectively. Larvae derived from gametes containing PbTx showed significantly higher mortalities and attained a smaller larval size for the first 6 days post-fertilization. These negative effects on larval development may be due to the presence of PbTx in the lipid droplets of the oocytes, which is mobilized by the larvae during embryonic and lecithotrophic larval development. Provision of a non-contaminated food source to larvae however, appeared to mitigate the early negative effects of this neonatal PbTx exposure. Results herein show that adult eastern oysters and their offspring are susceptible to exposure to K. brevis. Caution should therefore be exercised when identifying oyster reef restoration areas and in efforts to establish aquaculture in areas prone to red tides.

Susceptibility of gametes and embryos of the eastern oyster, Crassostrea virginica, to Karenia brevis and its toxins.

The bivalve mollusc, Crassostrea virginica, is frequently exposed to blooms of Karenia brevis along the west coast of Florida during periods of spawning and early larval development. A continuous 4-day exposure of gametes and 2-4 cell stage embryos of C. virginica to whole-cell and culture filtrate of K. brevis at 500 and 5000 cells mL(-1), was followed by a 4-day 'recovery' period. Larval growth, percent of normal, abnormal and dead larvae, and the presence of food in the larval gut were measured throughout the exposure period. Results suggest that negative effects mainly occur during embryogenesis and early development. Damage to feeding apparatus/gut may occur during embryonic development or exposure to toxins may act as a feeding deterrent on non-toxic algae. Following 2-h in vitro exposure of gametes, differences in oocyte and sperm cell parameters were investigated using flow cytometry. The reduced sperm viability in the whole-cell 5000 cells mL(-1) treatment suggests the involvement of extracellular brevetoxins (PbTx) and perhaps other harmful, uncharacterized compounds associated with the K. brevis cell membrane. The cumulative effects of reduced sperm viability, fertilization success, embryonic and larval survival, and the near-annual exposure to blooms of K. brevis could cause significant bottlenecks on oyster recruitment.

Effects of the red tide dinoflagellate, Karenia brevis, on early development of the eastern oyster Crassostrea virginica and northern quahog Mercenaria mercenaria.

The brevetoxin-producing dinoflagellate, Karenia brevis, adversely affects many shellfish species including the commercially and ecologically important bivalve molluscs, the northern quahog (=hard clam) Mercenaria mercenaria and eastern oyster Crassostrea virginica, in the Gulf of Mexico, USA. This study assessed the effects of exposure of these bivalves to K. brevis during their early development. In separate experiments, embryos of 2-4 cell stage of M. mercenaria and C. virginica were exposed to both whole and lysed K. brevis cells isolated from Manasota Key, Florida. Low bloom concentrations of 500 to 3000 cells mL(-1) were simulated for 96 h. Shell length, percent abnormality (and normality), and percent mortality of resulting larvae were measured. Percentages were recorded after 6, 24, and 96 h of exposure; larval shell length was measured at 24 and 96 h. For both quahogs and oysters, the effects of exposing embryos to K. brevis on all larval responses were generally dose- and time-dependent. Percent mortalities and abnormalities of both clam and oyster embryos increased significantly after only 6h of exposure to whole cells of K. brevis. For clams, these parameters were significantly higher in whole and lysed treatments (at 3000 cells mL(-1)) than in controls. Percent mortalities of oysters were significantly higher in the whole-cell treatment (3000 cells mL(-1)) than under control conditions. After 24h of exposure, mean larval shell length of both bivalve species was significantly reduced relative to controls. This was evident for clam larvae in both the lysed treatment at 1500 cells mL(-1) and in whole and lysed treatments at 3000 cells mL(-1), and for oyster larvae in the lysed treatment at 3000 cells mL(-1). After 96 h, both species exposed to the lysed cell treatment at 3000 cells mL(-1) had significantly smaller larvae compared to those in the control. Overall, lysed cells of K. brevis had a more pronounced effect on shell length, percent abnormality, and mortality in both clams and oysters than did whole cells. Given the fact that blooms of K. brevis overlap with the spawning periods of these two bivalves, and that cells of this naked dinoflagellate are readily lysed by wave action, these results suggest that exposure to K. brevis during the early life history stages of clams and oysters could adversely affect their population recruitment. Further, the presence of whole or lysed cells of K. brevis in hatcheries could have a major negative impact on production.