Toxic Dinoflagellate Gymnodinium Catenatum Research Articles

Resting Cysts of the Toxic Dinoflagellate Gymnodinium catenatum (Dinophyceae) Ubiquitously Distribute along the Entire Coast of China, with Higher Abundance in Bloom-Prone Areas

Blooms of Gymnodinium catenatum have occurred occasionally in different areas of China and caused tremendous economic loss and even threatened human health. Not only is G. catenatum an important harmful-algal-bloom (HAB)-causing species, but also the only gymnodinioid dinoflagellate known to produce paralytic shellfish poisoning toxins (PSTs). Due to the germination of resting cysts, which often initiates blooms, the distribution and abundance of cysts in sediments and particularly the confirmation of cyst beds are important information for understanding and predicting dinoflagellate blooms. In this research, 199 sediment samples were collected from China’s coastal seas, ranging from the Beidaihe in the Bohai Sea (BS) to the southernmost sample from the Nansha Islands of the South China Sea (SCS). TaqMan quantitative PCR (qPCR) assays with species-specific primers and probes were developed to specifically detect the distribution and abundance of cysts in the 199 samples. The detection revealed that G. catenatum cysts were widely present in the sediments (126 of the 199 samples), with 93.55%, 74.65%, 42.37%, and 50% of the samples detected positively from the BS, YS, ECS and SCS, respectively, and covering the vast sea area from Nansha Islands to the Beidaihe area. The single-cyst morpho-molecular identification in the samples from Beidaihe confirmed the existence of G. catenatum cysts in the BS, and the positive detections of G. catenatum cysts using the qPCR methods. While G. catenatum cysts were widely distributed in all four seas of China, the average abundance was relatively low (1.0 cyst per gram of wet sediment). Three samples from the East China Sea (ECS), however, contained G. catenatum cysts at a relatively higher level (23 cysts g−1 wet sediment) than other sea areas, suggesting a pertinence of cyst abundance to the frequent occurrences of G. catenatum blooms in the area during recent years. Collectively, for G. catenatum being such an important toxic and HAB-causing species globally, the ubiquitous distribution of its cysts along the coastal waters of China and higher abundance in the bloom-prone areas warns us of a risk that cyst beds, although currently low in abundance, may seed HABs in any and many sea areas of China at any forthcoming year, and particularly those areas with records of frequent HABs outbreaks in the past.

Changes in Toxin Production, Morphology and Viability of Gymnodinium catenatum Associated with Allelopathy of Chattonella marina var. marina and Gymnodinium impudicum.

Allelopathy between phytoplankton organisms is promoted by substances released into the marine environment that limit the presence of the dominating species. We evaluated the allelopathic effects and response of cell-free media of Chattonella marina var. marina and Gymnodinium impudicum in the toxic dinoflagellate Gymnodinium catenatum. Additionally, single- and four-cell chains of G. catenatum isolated from media with allelochemicals were cultured to evaluate the effects of post exposure on growth and cell viability. Cell diagnosis showed growth limitation and an increase in cell volume, which reduced mobility and led to cell lysis. When G. catenatum was exposed to cell-free media of C. marina and G. impudicum, temporary cysts and an increased concentration of paralytic shellfish toxins were observed. After exposure to allelochemicals, the toxin profile of G. catenatum cells in the allelopathy experiments was composed of gonyautoxins 2/3 (GTX2/3), decarcarbamoyl (dcSTX, dcGTX2/3), and the sulfocarbamoyl toxins (B1 and C1/2). A difference in toxicity (pg STXeq cell−1) was observed between G. catenatum cells in the control and those exposed to the filtrates of C. marina var. marina and G. impudicum. Single cells of G. catenatum had a lower growth rate, whereas chain-forming cells had a higher growth rate. We suggest that a low number of G. catenatum cells can survive the allelopathic effect. We hypothesize that the survival strategy of G. catenatum is migration through the chemical cloud, encystment, and increased toxicity.

Effects of paralytic shellfish poisoning toxin-producing dinoflagellate Gymnodinium catenatum on the marine copepod Tigriopus japonicus

To understand how the marine copepod Tigriopus japonicus responds to the toxic marine dinoflagellate Gymnodinium catenatum, we assessed acute toxicity and investigated swimming behavior parameters (e.g., swimming speed, swimming path trajectory, and swimming distance) in response to G. catenatum exposure. In addition, the mRNA expression levels of detoxification-related genes (e.g., phase I cytochrome P450 [CYP] and phase II glutathione-S transferase [GST]) were measured in G. catenatum-exposed copepods. No significant change in survival was observed in response to G. catenatum, but swimming speed was significantly decreased (P<0.05) at a high concentration of G. catenatum (600cells/mL). Furthermore, the swimming distance was significantly decreased (P<0.05) compared to that of the control at 600 cells/mLG. catenatum, while no significant change in swimming path trajectory was observed, suggesting that G. catenatum potentially has adverse effects on the swimming behavior of T. japonicus. In addition, the transcriptional regulation of T. japonicus CYPs and -GSTs were significantly upregulated and downregulated (P<0.05), respectively, in response to G. catenatum. In particular, certain genes (e.g., CYPs [CYP307E1, CYP3041A1, and CYP3024A2] and GSTs [GST-kappa, GST-mu5, and GST-omega]) were significantly induced (P<0.05) by G. catenatum, suggesting that these genes likely play a critical role in detoxification mechanisms and might be useful as potential molecular biomarkers in response to G. catenatum exposure. Overall, these results elucidate the potential impacts of the dinoflagellate G. catenatum on the swimming behavior and detoxification system of the marine copepod T. japonicus.

Effects of Different Nutrient and Trace Metal Concentrations on Growth of the Toxic Dinoflagellate Gymnodinium catenatum Isolated from Korean Coastal Waters

The effects of the addition of nutrients (nitrate: N; phosphate: P; and vitamin B1) and trace metals (iron: Fe; Copper: Cu; and selenium: Se) on the growth of Gymnodinium catenatum, which was isolated from Korean coastal waters, were investigated. The Korean isolate of G. catenatum grew under a wide range of concentrations of N and P. Whilst high concentrations of N (&gt; N: P ratio of 23.5) did not stimulate the growth rate, an enhanced growth rate and cell density were observed with the addition of P. The experimental addition of vitamin B1 revealed that G. catenatum is not dependent on vitamin B1 for growth. Moreover, the addition of Fe and Cu resulted in no significant differences in the growth patterns and rates of G. catenatum between the controls and treatments. It is thus possible that growth of the Korean isolate of G. catenatum does not require high concentrations of Fe and Cu. However, the cell densities were enhanced in the stationary phases of treatments upon addition of Se, and the maximum cell densities were higher than those in the culture experiments upon additions of other nutrient and trace metals. Our findings indicate that G. catenatum prefers P and Se for proliferation, rather than other nutritional sources.

Plankton community and copepod production in a temperate coastal lagoon: What is changing in a short temporal scale?

Coastal lagoons are often exposed to intense short-term environmental changes and strong anthropogenic pressures influencing zooplanktonic communities and production. However, most works focus on long-term temporal scales using monthly or seasonal sampling strategies. The present study analysed the structure of the mesozooplanktonic assemblages, the production (egg production rates) and physiological condition (RNA:DNA ratio) of the copepod Acartia clausi in a temperate coastal lagoon (Ria Formosa) during the summer, using an intensive sampling approach. Salinity was the main factor affecting the short-term variability of mesozooplankton composition, followed by tidal phase (ebb tides) and semilunar cycle (spring tides). There was a positive relationship between the abundance of Appendicularia and the cladoceran Penilia avirostris with the toxic dinoflagellate Gymnodinium catenatum, suggesting no deleterious effects. The egg production rate of A. clausi was influenced by salinity and ammonia concentration, with a positive correlation between the egg productivity and the macronutrient, showing a possible adaptation of this calanoid species. The RNA:DNA index was positively related to egg production rate, suggesting that it is a good proxy for the reproductive output of copepods, even in short-term periods. This study shows that different timescales need to be included in regular monitoring of planktonic assemblages in coastal lagoons in order to understand the influence of environmental and anthropogenic variables on marine organisms.

Bacterial Associates Modify Growth Dynamics of the Dinoflagellate Gymnodinium catenatum.

Marine phytoplankton cells grow in close association with a complex microbial associate community known to affect the growth, behavior, and physiology of the algal host. The relative scale and importance these effects compared to other major factors governing algal cell growth remain unclear. Using algal-bacteria co-culture models based on the toxic dinoflagellate Gymnodinium catenatum, we tested the hypothesis that associate bacteria exert an independent effect on host algal cell growth. Batch co-cultures of G. catenatum were grown under identical environmental conditions with simplified bacterial communities composed of one-, two-, or three-bacterial associates. Modification of the associate community membership and complexity induced up to four-fold changes in dinoflagellate growth rate, equivalent to the effect of a 5°C change in temperature or an almost six-fold change in light intensity (20–115 moles photons PAR m-2 s-1). Almost three-fold changes in both stationary phase cell concentration and death rate were also observed. Co-culture with Roseobacter sp. DG874 reduced dinoflagellate exponential growth rate and led to a more rapid death rate compared with mixed associate community controls or co-culture with either Marinobacter sp. DG879, Alcanivorax sp. DG881. In contrast, associate bacteria concentration was positively correlated with dinoflagellate cell concentration during the exponential growth phase, indicating growth was limited by supply of dinoflagellate-derived carbon. Bacterial growth increased rapidly at the onset of declining and stationary phases due to either increasing availability of algal-derived carbon induced by nutrient stress and autolysis, or at mid-log phase in Roseobacter co-cultures potentially due to the onset of bacterial-mediated cell lysis. Co-cultures with the three bacterial associates resulted in dinoflagellate and bacterial growth dynamics very similar to more complex mixed bacterial community controls, suggesting that three-way co-cultures are sufficient to model interaction and growth dynamics of more complex communities. This study demonstrates that algal associate bacteria independently modify the growth of the host cell under non-limiting growth conditions and supports the concept that algal–bacterial interactions are an important structuring mechanism in phytoplankton communities.

Paralytic shellfish toxins, including deoxydecarbamoyl-STX, in wild-caught Tasmanian abalone (Haliotis rubra)

For the first time wild-caught Tasmanian abalone, Haliotis rubra, have been reported to contain paralytic shellfish toxins (PSTs). This observation followed blooms of the toxic dinoflagellate Gymnodinium catenatum. No illnesses were reported, but harvesting restrictions were enforced in commercial areas. Abalone were assayed using HPLC-FLD methodology based on AOAC official method 2005.06. An uncommon congener, deoxydecarbamoyl-STX (doSTX), was observed in addition to regulated PSTs as unassigned chromatographic peaks. A quantitative reference material was prepared from contaminated Tasmanian abalone viscera and ampouled at 54.2 μmol/L. The LD50 of doSTX via intraperitoneal injection was 1069 nmol/kg (95% confidence limits 983–1100 nmol/kg), indicating it is nearly 40 times less toxic than STX. A toxicity equivalence factor of 0.042 was generated using the mouse bioassay.Levels of PSTs varied among individuals from the same site, although the toxin profile remained relatively consistent. In the foot tissue, STX, decarbamoyl-STX and doSTX were identified. On a molar basis doSTX was the dominant congener in both foot and viscera samples. The viscera toxin profile was more complex, with other less toxic PST congeners observed and was similar to mussels from the same site. This finding implicates localised dinoflagellate blooms as the PST source in Tasmanian abalone.

Apoptosis of hemocytes from lions-paw scallop Nodipecten subnodosus induced with paralyzing shellfish poison from Gymnodinium catenatum

The toxic dinoflagellate Gymnodinium catenatum produces paralyzing shellfish poisons (PSPs) that are consumed and accumulated by bivalves. Previously, we recorded a decrease in hemocytes 24h after injection of PSPs (gonyautoxin 2/3 epimers, GTX2/3) in the adductor muscle in the lions-paw scallop Nodipecten subnodosus. In this work, qualitative and quantitative analyses, in in vivo and in vitro experiments, revealed that the lower count of hemocytes results from cells undergoing typical apoptosis when exposed to GTX 2/3 epimers. This includes visible morphological alterations of the cytoplasmic membrane, damage to the nuclear membrane, condensation of chromatin, DNA fragmentation, and release of DNA fragments into the cytoplasm. Induction of apoptosis was accompanied by phosphatidylserine exposure to the outer cell membrane and activation of cysteine-aspartic proteases, caspase 3 and caspase 8. Addition of an inhibitor of caspase to the medium suppressed activation in hemocytes exposed to the toxins, suggesting that cell death was induced by a caspase-dependent apoptotic pathway. The results are important for future investigation of the scallop's immune system and should provide new insights into apoptotic processes in immune cells of scallops exposed to PSPs.

Deterrent effect of Gymnodinium catenatum Graham PSP-toxins on grazing performance of marine copepods

The present study aimed to determine the effects of different concentrations of the toxic dinoflagellate Gymnodinium catenatum on the feeding performance, survival rates, and PSP toxin profiles of Euterpina acutifrons Dana and Acartia grani Sars. Each of the three different experiments was conducted in order to determine the feeding rates of adult female copepod species. Feeding experiments were performed in four 75-ml vessels each containing five copepods and different concentrations of cells (C1=2.68×102, C2=5.36×102cellsml−1). Initial and final cell concentrations were estimated using a Coulter counter, while the carbon and nitrogen content were determined using a CHN analyzer. High-performance liquid chromatography was used to determine the composition of the PSP toxins, and estimate their concentrations in the tissue of the copepods. In experiment 1, significantly higher (p<0.05) ingestion rates were recorded for A. grani when feeding on the second concentration of cells (C2). Filtration rates were also significantly higher (p<0.05) for C2. A similar pattern was recorded for ingestion rates in E. acutifrons. By contrast, filtration rates were apparently higher for the lower concentration (C1). In experiment 2, A. grani ingestion rates were significantly higher (p<0.05) for the higher concentration (C2) in all three time phases. However, filtration rates for C2 were significantly higher (p<0.05) at 12 and 18h. E. acutifrons also presented significantly higher ingestion rates (p<0.05) for C2 at 12 and 18h. Filtration rates were significantly higher (p<0.05) for C2 only in the 12-h phase. In experiment 3, both copepods presented significantly lower (p<0.05) ingestion and filtration rates following exposure to G. catenatum for 24h. Carbamoyl toxins in the chromatograms were dominated by GTX1 and STX, followed by GTX2 and GTX3. Total cell toxicity was 2.58 and 2.34pgSTXeqcell−1 for cells harvested at the beginning and the end of the experiments, respectively. Total toxicity in the tissues of E. acutifrons and A. grani was 11.51 and 7.56(gSTXeqg−1, respectively. Survival rates in A. grani and E. acutifrons fed on C2 were 0% after 168 and 192h, respectively. Overall, the present study indicates that the copepods E. acutifrons and A. grani can not only ingest G. catenatum toxic cells, but also transfer their PSP toxins to other trophic levels. However, G. catenatum PSP toxins were shown to have had a deterrent and negative time-dependent effect on the feeding and survival rates, respectively, of E. acutifrons and A. grani.

Uptake and release of paralytic shellfish toxins by the clam Ruditapes decussatus exposed to Gymnodinium catenatum and subsequent depuration

A laboratory experiment was performed with the clam Ruditapes decussatus, fed with the toxic dinoflagellate Gymnodinium catenatum and the non-toxic algae Isochrysis galbana (14 days) and subsequently only with I. galbana (15 days). Individual paralytic shellfish toxins were determined by LC-FLD in G. catenatum cells, whole clam tissues, and particulate organic matter (POM) produced by clams. The toxins dcSTX and dcGTX2 + 3 in the algae were less abundant than C1 + 2 and B1, but were predominant in clams during both the exposure and depuration phases. The toxin dcNEO was only detected in clams during a short period, indicating conversion from other compounds. The toxin composition of the POM indicated the export of dcSTX as faeces or pseudo-faeces along the entire experiment (2.5–14 nmol mg−1), B1 was present in a short period of the exposure and C1 + 2 and dcGTX2 + 3 absent. A mass balance calculation indicated that approximately 95% of C1 + 2 and 85% of B1 supplied to the clams were converted into other toxins or lost in solution. Conversely, the net gain of 512, 61 and 31 nmol for dcSTX, dcGTX2 + 3 and dcNEO, respectively, suggests the conversion from other assimilated compounds by clams during exposure and depuration phases.

THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM (DINOPHYCEAE) REQUIRES MARINE BACTERIA FOR GROWTH(1).

Interactions with the bacterial community are increasingly considered to have a significant influence on marine phytoplankton populations. Here we used a simplified dinoflagellate-bacterium experimental culture model to conclusively demonstrate that the toxic dinoflagellate Gymnodinium catenatum H. W. Graham requires growth-stimulatory marine bacteria for postgermination survival and growth, from the point of resting cyst germination through to vegetative growth at bloom concentrations (10(3) cells · mL(-1) ). Cysts of G. catenatum were germinated and grown in unibacterial coculture with antibiotic-resistant or antibiotic-sensitive Marinobacter sp. DG879 or Brachybacterium sp., and with mixtures of these two bacteria. Addition of antibiotics to cultures grown with antibiotic-sensitive strains of bacteria resulted in death of the dinoflagellate culture, whereas cultures grown with antibiotic-resistant bacteria survived antibiotic addition and continued to grow beyond the 21 d experiment. Removal of either bacterial type from mixed-bacterial dinoflagellate cultures (using an antibiotic) resulted in cessation of dinoflagellate growth until bacterial concentration recovered to preaddition concentrations, suggesting that the bacterial growth factors are used for dinoflagellate growth or are labile. Examination of published reports of axenic dinoflagellate culture indicate that a requirement for bacteria is not universal among dinoflagellates, but rather that species may vary in their relative reliance on, and relationship with, the bacterial community. The experimental model approach described here solves a number of inherent and logical problems plaguing studies of algal-bacterium interactions and provides a flexible and tractable tool that can be extended to examine bacterial interactions with other phytoplankton species.

Effects of the toxic dinoflagellate, Gymnodinium catenatum on hydrolytic and antioxidant enzymes, in tissues of the giant lions-paw scallop Nodipecten subnodosus

This study documents effects of the toxic dinoflagellate Gymnodinium catenatum, a producer of paralytic shellfish poison, on juvenile farmed (5.9 ± 0.39 cm) giant lions-paw scallop Nodipecten subnodosus. Scallops were fed bloom concentrations of toxic dinoflagellate G. catenatum for 7 h. The effect of the toxic dinoflagellate in different tissues was determined by analysis of antioxidant enzymes (catalase, superoxide dismutase, gluthathione peroxidase), thiobarbituric acid reactive substances (lipid peroxidation), and hydrolytic enzymes (proteases, glycosidases, phosphatases, lipases, and esterases). Histopathological photos record the effects of the toxic dinoflagellate in various tissues. The results show that juvenile lions-paw scallops produce pseudo-feces, partially close their shell, increase melanization, and aggregate hemocytes. Several enzymes were affected and could serve as biological markers. In general, the adductor muscle was not affected. In the digestive gland, some enzymes could be the result of defensive and digestive processes. Gills and mantle tissue were markedly affected because these sites respond first to toxic dinoflagellates, leading to the idea that proteolytic cascades could be involved.

Widespread presence of hydrophobic paralytic shellfish toxins in Gymnodinium catenatum

The toxic dinoflagellate Gymnodinium catenatum Graham produces a newly discovered sub-class of paralytic shellfish toxins (PSTs, saxitoxins) that contain a hydroxybenzoate moiety in place of the carbamoyl group (GC toxins: GC1–GC3). GC toxins bind strongly to sodium channels and their lipophilic nature may increase their potential to bioaccumulate in marine organisms. Cultures Australian G. catenatum strains were found to contain 12–63 mol% GC toxins. The GC toxins were also detected in strains from China (38 mol%), Japan (1–2 mol%), Portugal (58 mol%), Spain (36–54 mol%), and Uruguay (10–16 mol%). A cluster analysis of molar proportions of saxitoxin derivatives produced by strains showed clear clustering by country/region of origin, indicating that GC toxins may be very useful markers to identify the source of G. catenatum in the case of new outbreaks. The GC toxins dominate the toxin profiles of many G. catenatum strains, and can contribute significantly to sample toxicity, yet these toxins may easily escape detection using conventional chromatography, resulting in significant underestimates of sample toxicity. This has significant implications for shellfish monitoring and safety.

Reflections on the ballast water dispersal—harmful algal bloom paradigm

The ballast water dispersal—HAB paradigm, increasingly invoked circumstantially to explain puzzling and unaccountable HAB species outbreaks when lacking the multiple tests of confirmation recommended by Bolch and de Salas (2007), is evaluated. The types and examples of natural dispersions and taxon cycles are compared to exotic species bloom behavior linked to ballast water vectoring. The regional spreading, bloom behavior and disjunct distributions of the brown tide pelagophyte Aureococcus anophagefferens and the toxic dinoflagellate Gymnodinium catenatum, attributed to ballast water vectoring, are used as representative examples to evaluate the general application of the ballast water—HAB paradigm and associated interpretative problems. Human-aided emigration has a seeding and colonization ecology that differs from bloom ecology. For self-sustaining blooms to occur, these two ecologies must be accommodated by habitat growth conditions. The three stages that a non-native species must pass through (pioneering, persistence, community entry) to achieve colonization, community maintenance, and to bloom, and the niche-related factors and role of habitat disturbance are discussed. The relevance of cryptic occurrences, cyst deposits, dormancy periods and bloom rhythms of HAB species to their blooms attributed to ballast water-assisted introductions is also sketched. The different forms of HAB species rarity, their impact on the ballast water dispersal—HAB paradigm, and the dispersion and blooms of specialist and generalist HAB species are discussed. The remarkable novel and, often, monospecific blooms of dinoflagellate HAB species are being paralleled by similar eruptive bloom behavior cutting across phylogenetic lines, and being found also in raphidophytes, haptophytes, diatoms, silicoflagellates, etc. These blooms cannot be explained only as seeding events. An ecological release of ‘old barriers’ appears to be occurring generally at coastal bloom sites, i.e. something significant is happening ecologically and embedded within the ballast water—HAB paradigm. There may be a relationship between Life Form type [Smayda, T.J., Reynolds, C.S., 2001. Community assembly in marine phytoplankton: application of recent models to harmful dinoflagellate blooms. J. Plankton Res. 23, 447–461] and mode of expatriation; HAB dinoflagellate species commonly reported to produce ballast water-assisted toxic blooms invariably are members of cyst-producing Life Forms IV, V, VI. Ballast water vectoring of Life Forms I, II, III is rarely reported, even though many produce cysts, and where their novel introductions do occur they are more likely to be ichthyotoxic and vectored in shellfish stock consignments. The relevance of, and need to distinguish between morphospecies and their geographic/ribotype clades are discussed based on the Alexandrium tamarense/catenella/fundyense complex. Morphospecies-level ballast water dispersions are probably minor compared to the dispersal of the different ribotypes (toxic/non-toxic clades) making up HAB morphospecies; the redistribution and admixture of genotypes should be the focus. Ballast water-assisted expatriations impact the global occurrence of HABs through the direct transfer of previously absent species or introduction of genetic strains from the donor habitat that are ecologically favored over resident strains. The hybridization of species may be of potentially greater impact, resulting from the (1) mating of individuals from the donor and recipient habitats, or (2) through the interbreeding of strains introduced from two different donor sites into the recipient site, and whose progeny have greater ecological fitness than indigenous strains. Exceptional ecological changes of some sort appear to be occurring globally which, in combination with the genetically altered ecophysiological behavior of HAB species linked to ballast water dispersion and admixture, underpins the global HAB phenomenon. The impact of ballast water and shellfish transplantation on HABs and phytoplankton community ecology, generally, is considerably greater than the current focus on HAB species distributions, vectoring, and blooms. The methodological, investigative and conceptual potential of the ballast water—HAB paradigm should be exploited by developing a GEOHAB type intiative to advance quantification of global HAB ecology.

Accumulation and Elimination Profiles of Paralytic Shellfish Poison in the Short-necked Clam Tapes japonica Fed with the Toxic Dinoflagellate Gymnodinium catenatum

The paralytic shellfish poison (PSP)-producing dinoflagellate Gymnodinium catenatum (Gc) was fed to the short-necked clam Tapes japonica, and the accumulation, transformation and elimination profiles of PSP were investigated by means of high-performance liquid chromatography with postcolumn fluorescence derivatization (HPLC-FLD). The short-necked clams ingested most of the Gc cells (4 x 10(6) cells) supplied as a bolus at the beginning of the experiment, and accumulated a maximal amount of toxin (181 nmol/10 clams) after 12 hr. The rate of toxin accumulation at that time was 16%, which rapidly decreased thereafter. During the rearing period, a variation in toxin composition, derived presumably from the transformation of toxin analogues in the clams, was observed, including a reversal of the ratio of C2 to C1, and the appearance of carbamate (gonyautoxin (GTX) 2, 3) and decarbamoyl (dc) derivatives (decarbamoylsaxitoxin (dcSTX) and dcGTX2, 3), which were undetectable in Gc cells. The total amount of toxin contained in clams and residue (remaining Gc cells and/or excrement in the rearing tank) gradually declined, and only about 1% of the supplied toxin was detected at the end of the experiment.

Effects of the toxic dinoflagellate Gymnodinium catenatum on uptake and fate of paralytic shellfish poisons in the Pacific giant lions-paw scallop Nodipecten subnodosus

Juvenile Pacific giant lions-paw scallops Nodipecten subnodosus were fed the toxic dinoflagellate Gymnodinium catenatum, a producer of paralytic shellfish poison (PSP), supplied with Isochrysis galbana (a nontoxic microalgae). Short-term ( 2 days). At high food concentrations, juvenile scallops showed production of pseudofeces, partial shell valve closure, and reduction in feeding. According to HPLC analysis, the only toxins present in the dinoflagellate G. catenatum and in the scallops were the gonyautoxins (GTXs), except in the labial palps and digestive gland, where trace amounts of saxitoxin (STX) were present in scallops. These tissues could play an important role in toxin biotransformation. The ranking of toxin concentration in tissues was: digestive gland > labial palps > intestine > gills > mantle > adductor muscle, where the total contribution of viscera was more than 80% of the total toxin body burden. Juvenile scallops exhibited no apparent detrimental physiological responses during the long-term feeding experiment. The dinoflagellate may contribute nutrients to the scallop, in addition to the microalgae I. galbana. The dinoflagellate may enhance cell uptake and byssus production. Once PSP accumulated during the first 12 days, it was slowly eliminated. The limited capacity for accumulating toxins in the adductor muscle favors domestic marketing of scallops.

Vertical migration of the toxic dinoflagellate Gymnodinium catenatum under different concentrations of nutrients and humic substances in culture

Vertical migration behaviour by the chainforming dinoflagellate Gymnodinium catenatum Graham was investigated using vertically-stratified laboratory columns. Under surface nutrient-deplete conditions, with nutrients added only at depth, 100% of cells underwent vertical migration (VM), starting downwards migration 3 h before the end of the light period and beginning upwards migration 3 h before the start of the light period. Cells in nutrient-replete columns showed no VM, but they were more dispersed in the upper layer during the dark compared to the light period. When surface layers (S) were nitrate-deplete (−N) and enriched with humic substances (H) contained in Huon River water and bottom waters (B) were nutrient replete (R) (SH −NB R), the pattern of VM was altered—50% of cells underwent migration and 50% remained at the pycnocline. In columns with nitrate-replete and humic-enriched surface layers (SH RB R), most cells underwent VM, while 30% remained at the surface. Cells in SH −NB R columns showed increased N quotas and intra-cellular nitrate concentrations after 4 days, indicating nitrate uptake by G. catenatum in bottom layers. The concomitant increase in particulate organic nitrogen (PON) with the decrease in external nitrate concentrations in bottom layers provide convincing evidence that VM by G. catenatum facilitates nutrient retrieval at depth. However, addition of humic substances (a potential source of organic nitrogen) to surface layers did not ameliorate G. catenatum N depletion sufficiently to preclude the need for NO 3 − uptake at depth. Furthermore, there was no detectable pattern of increasing carbon (C) quota during the day (photosynthate accumulation) or increasing N quota during the night (nitrate assimilation). Toxic dinoflagellate G. catenatum blooms are commonly associated with nitrate depletion in surface waters in south-east Tasmanian waters (Australia). Therefore, vertical migration, facilitating N uptake at depth, could play an important role in this organism's ecological strategy, enabling it to exploit environments where light and nutrients are vertically separated.

Growth and uptake kinetics for nitrate, ammonium and phosphate by the toxic dinoflagellate Gymnodinium catenatum isolated from Hiroshima Bay, Japan

: Growth and uptake kinetics for nitrate, ammonium and phosphate by Gymnodinium catenatum isolated from Hiroshima Bay were experimentally investigated. Maximum uptake rate (ρmax) and half saturation constant (Ks), which were obtained by short-term uptake experiments, were 6.48 pmol/cell per h and 7.60 µM for nitrate, 3.37 pmol/cell per h and 33.6 µM for ammonium, and 1.42 pmol/cell per h and 3.40 µM for phosphate, respectively. Specific maximum growth rate (µ′m) and minimum cell quota (Q0), which were obtained by semicontinuous growth experiments, were 0.36/day and 31.3 pmol/cell for the N-limited conditions, respectively. The µ′m and QP0 for P-limited conditions were 0.37/day and 1.83 pmol/cell, respectively. These results indicate that G. catenatum is a poor competitor in terms of utilization of inorganic nutrients and is unlikely to form blooms in Hiroshima Bay where phosphate is chronically depleted unless the growth is supported by organic species (i.e. dissolved organic phosphorus) of the nutrients.

Properties of light absorption in a highly coloured estuarine system in south-east Australia which is prone to blooms of the toxic dinoflagellate Gymnodinium catenatum

The Huon Estuary in south-east Tasmania is an important site in Australia's aquaculture and finfish industries. Atlantic salmon and shellfish are farmed in the Huon River Estuary, which drains a catchment that includes both areas of pristine wilderness and agriculture. The estuary experiences algal blooms, including blooms of the toxic dinoflagellate Gymnodinium catenatum, which can cause considerable problems for the aquaculture industry. The freshwater input is highly coloured due to high levels of humic material and this combined with intrusions of clearer oceanic water and the occurrence of algal blooms makes this estuary optically complex. Between November 1996 and September 1998 samples for pigment and optical analyses were collected weekly from five sites within the mid to lower regions of the estuary. In addition, every three months (beginning July 1996), samples were collected from 30 sites within the full river/estuary system. Early in December 1997 a bloom of Gymnodinium catenatum occurred throughout the estuary and persisted until June 1998. Chromophoric dissolved organic matter (CDOM) was found to dominate the total absorption of the water throughout the entire estuary and over the two-year duration of the study. The occurrence of algal blooms showed no correlation with the optical characteristics of the estuary, suggesting that optical parameters cannot be used to predict the occurrence of algal blooms in this estuary. Blooms of different algal species, such as diatoms and dinoflagellates, may be able to be distinguished by their absorption spectra in the UV region rather than the visible region. To date, this study is the most detailed spatial and temporal study of the characteristics of light absorption in an estuarine system within Australia.

Diatom and dinoflagellate assemblages of the Hawkesbury River, N.S.W., over the last two centuries: evidence for changes in hydrology

Diatom and dinoflagellate cyst analysis of a 77 cm long sediment core from Cowan Creek, Hawkesbury River estuary, N.S.W., revealed changes in the catchment hydrology over the last 266 years. High abundances of the freshwater/brackish diatom genus Cyclotella at the base of the core imply sustained periods of reduced salinity that now no longer occur. Reduction of freshwater flow after approximately circa 1800 (60 cm) has allowed the development of marine planktonic diatoms Thalassiosira spp.,Ditylum brightwellii, Rhizosolenia setigera, Pseudo-nitzschia pungens and Chaetoceros spp. Benthic diatom diversity has remained relatively unchanged. The toxic dinoflagellate Gymnodinium catenatum, although identified in a cyst survey in April 1995, was not found in the sediment cores. Changes in dinoflagellate assemblage are consistent with the effects of increasing urbanisation and eutrophication.