Dinoflagellate Cells Research Articles

Seasonal variation in composition and abundance of harmful dinoflagellates in Yemeni waters, southern Red Sea

General abundance and species composition of a dinoflagellate community in Yemeni coastal waters of Al Salif (southern Red Sea) were studied with a view to understand the annual variations in particular the toxic species. Dinoflagellates were more abundant among phytoplankton. Thirty five dinoflagellate taxa were identified, among which 12 were reported as potentially toxic species. A significant change in seasonal abundance was recorded with the maximum (2.27∗106cellsl−1) in May, and the minimum (2.50∗102cellsl−1) recorded in January. Kryptoperidinium foliaceum, which was reported for the first time from the Red Sea, was the most abundant species with a maximum in May 2013 (2.26∗106cellsl−1). Spearman's rank correlation analysis indicates that, total harmful dinoflagellate cells, K. foliaceum, Prorocentrum gracile and Prorocentrum micans were significantly correlated with temperature. This study suggests that Yemeni waters should be monitored to investigate harmful species and to identify areas and seasons at higher risk.

Quantitative Determination and Subcellular Imaging of Cu in Single Cells via Laser Ablation-ICP-Mass Spectrometry Using High-Density Microarray Gelatin Standards.

This manuscript describes the development and characterization of a high-density microarray calibration standard, manufactured in-house and designed to overcome the limitations in precision, accuracy, and throughput of current calibration approaches for the quantification of elemental concentrations on the cellular level using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICPMS). As a case study, the accumulation of Cu in the model organism Scrippsiella trochoidea resulting from transition metal exposure (ranging from 0.5 to 100 μg/L) was evaluated. After the Cu exposure, cells of this photosynthetic dinoflagellate were treated with a critical point drying protocol, transferred to a carbon stub, and sputter-coated with a Au layer for scanning electron microscopy (SEM) analysis. In subsequent LA-ICPMS analysis, approximately 100 cells of each population were individually ablated. This approach permitted the evaluation of the mean concentration of Cu in the cell population across different exposure levels and also allowed the examination of the cellular distribution of Cu within the populations. In a cross-validation exercise, subcellular LA-ICPMS imaging was demonstrated to corroborate synchrotron radiation confocal X-ray fluorescence (SR-XRF) microimaging of single cells investigated under in vivo conditions.

The Vitamin B1 and B12 Required by the Marine Dinoflagellate Lingulodinium polyedrum Can be Provided by its Associated Bacterial Community in Culture.

In this study we established the B1 and B12 vitamin requirement of the dinoflagellate Lingulodinium polyedrum and the vitamin supply by its associated bacterial community. In previous field studies the B1 and B12 demand of this species was suggested but not experimentally verified. When the axenic vitamin un-supplemented culture (B-ns) of L. polyedrum was inoculated with a coastal bacterial community, the dinoflagellate’s vitamin growth limitation was overcome, reaching the same growth rates as the culture growing in vitamin B1B7B12-supplemented (B-s) medium. Measured B12 concentrations in the B-s and B-ns cultures were both higher than typical coastal concentrations and B12 in the B-s culture was higher than in the B-ns culture. In both B-s and B-ns cultures, the probability of dinoflagellate cells having bacteria attached to the cell surface was similar and in both cultures an average of six bacteria were attached to each dinoflagellate cell. In the B-ns culture the free bacterial community showed significantly higher cell abundance suggesting that unattached bacteria supplied the vitamins. The fluorescence in situ hybridization (FISH) protocol allowed the quantification and identification of three bacterial groups in the same samples of the free and attached epibiotic bacteria for both treatments. The relative composition of these groups was not significantly different and was dominated by Alphaproteobacteria (>89%). To complement the FISH counts, 16S rDNA sequencing targeting the V3–V4 regions was performed using Illumina-MiSeq technology. For both vitamin amendments, the dominant group found was Alphaproteobacteria similar to FISH, but the percentage of Alphaproteobacteria varied between 50 and 95%. Alphaproteobacteria were mainly represented by Marivita sp., a member of the Roseobacter clade, followed by the Gammaproteobacterium Marinobacter flavimaris. Our results show that L. polyedrum is a B1 and B12 auxotroph, and acquire both vitamins from the associated bacterial community in sufficient quantity to sustain the maximum growth rate.

Taxonomic and Environmental Variability in the Elemental Composition and Stoichiometry of Individual Dinoflagellate and Diatom Cells from the NW Mediterranean Sea.

Here we present, for the first time, the elemental concentration, including C, N and O, of single phytoplankton cells collected from the sea. Plankton elemental concentration and stoichiometry are key variables in phytoplankton ecophysiology and ocean biogeochemistry, and are used to link cells and ecosystems. However, most field studies rely on bulk techniques that overestimate carbon and nitrogen because the samples include organic matter other than plankton organisms. Here we used X-ray microanalysis (XRMA), a technique that, unlike bulk analyses, gives simultaneous quotas of C, N, O, Mg, Si, P, and S, in single-cell organisms that can be collected directly from the sea. We analysed the elemental composition of dinoflagellates and diatoms (largely Chaetoceros spp.) collected from different sites of the Catalan coast (NW Mediterranean Sea). As expected, a lower C content is found in our cells compared to historical values of cultured cells. Our results indicate that, except for Si and O in diatoms, the mass of all elements is not a constant fraction of cell volume but rather decreases with increasing cell volume. Also, diatoms are significantly less dense in all the measured elements, except Si, compared to dinoflagellates. The N:P ratio of both groups is higher than the Redfield ratio, as it is the N:P nutrient ratio in deep NW Mediterranean Sea waters (N:P = 20–23). The results suggest that the P requirement is highest for bacterioplankton, followed by dinoflagellates, and lowest for diatoms, giving them a clear ecological advantage in P-limited environments like the Mediterranean Sea. Finally, the P concentration of cells of the same genera but growing under different nutrient conditions was the same, suggesting that the P quota of these cells is at a critical level. Our results indicate that XRMA is an accurate technique to determine single cell elemental quotas and derived conversion factors used to understand and model ocean biogeochemical cycles.

Effects of acute ocean acidification on spatially-diverse polar pelagic foodwebs: Insights from on-deck microcosms

The polar oceans are experiencing some of the largest levels of ocean acidification (OA) resulting from the uptake of anthropogenic carbon dioxide (CO2). Our understanding of the impacts this is having on polar marine communities is mainly derived from studies of single species in laboratory conditions, while the consequences for food web interactions remain largely unknown. This study carried out experimental manipulations of natural pelagic communities at different high latitude sites in both the northern (Nordic Seas) and southern hemispheres (Scotia and Weddell Seas). The aim of this study was to identify more generic responses and achieve greater experimental reproducibility through implementing a series of short term (4d), multilevel (3 treatment) carbonate chemistry manipulation experiments on unfiltered natural surface-ocean communities, including grazing copepods. The experiments were successfully executed at six different sites, covering a diverse range of environmental conditions and differing plankton community compositions. The study identified the interaction between copepods and dinoflagellate cell abundance to be significantly altered by elevated levels of dissolved CO2 (pCO2), with dinoflagellates decreasing relative to ambient conditions across all six experiments. A similar pattern was not observed in any other major phytoplankton group. The patterns indicate that copepods show a stronger preference for dinoflagellates when in elevated pCO2 conditions, demonstrating that changes in food quality and altered grazing selectivity may be a major consequence of ocean acidification. The study also found that transparent exopolymeric particles (TEP) generally increased when pCO2 levels were elevated, but the response was dependent on the exact set of environmental conditions. Bacteria and nannoplankton showed a neutral response to elevated pCO2 and there was no significant relationship between changes in bacterial or nannoplankton abundance and that of TEP concentrations. Overall, the study illustrated that, although some similar responses exist, these contrasting high latitude surface ocean communities are likely to show different responses to the onset of elevated pCO2.

Quantity of the dinoflagellate sxtA4 gene and cell density correlates with paralytic shellfish toxin production in Alexandrium ostenfeldii blooms

Many marine dinoflagellates, including several species of the genus Alexandrium, Gymnodinium catenatum, and Pyrodinium bahamense are known for their capability to produce paralytic shellfish toxins (PST), which can cause severe, most often food-related poisoning. The recent discovery of the first PST biosynthesis genes has laid the foundation for the development of molecular detection methods for monitoring and study of PST-producing dinoflagellates. In this study, a probe-based qPCR method for the detection and quantification of the sxtA4 gene present in Alexandrium spp. and Gymnodinium catenatum was designed. The focus was on Alexandrium ostenfeldii, a species which recurrently forms dense toxic blooms in areas within the Baltic Sea. A consistent, positive correlation between the presence of sxtA4 and PST biosynthesis was observed, and the species was found to maintain PST production with an average of 6 genomic copies of sxtA4. In August 2014, A. ostenfeldii populations were studied for cell densities, PST production, as well as sxtA4 and species-specific LSU copy numbers in Föglö, Åland, Finland, where an exceptionally dense bloom, consisting of 6.3×106cellsL−1, was observed. Cell concentrations, and copy numbers of both of the target genes were positively correlated with total STX, GTX2, and GTX3 concentrations in the environment, the cell density predicting toxin concentrations with the best accuracy (Spearman's ρ=0.93, p<0.01). The results indicated that all A. ostenfeldii cells in the blooms harbored the genetic capability of PST production, making the detection of sxtA4 a good indicator of toxicity.

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 cellsmL−1 were simulated for 96h. Shell length, percent abnormality (and normality), and percent mortality of resulting larvae were measured. Percentages were recorded after 6, 24, and 96h of exposure; larval shell length was measured at 24 and 96h. 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 cellsmL−1) than in controls. Percent mortalities of oysters were significantly higher in the whole-cell treatment (3000 cellsmL−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 cellsmL−1 and in whole and lysed treatments at 3000 cellsmL−1, and for oyster larvae in the lysed treatment at 3000 cellsmL−1. After 96h, both species exposed to the lysed cell treatment at 3000 cellsmL−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.

Large-scale bloom of Akashiwo sanguinea in the Northern California current system in 2009

Significant seabird mortality on the Oregon (OR) and Washington (WA) coast in 2009 has been attributed to a massive bloom of the dinoflagellate Akashiwo sanguinea (K. Hirasaka) G. Hansen & Ø. Moestrup. Initial, albeit limited, observations suggested this bloom began in WA and reached OR waters through southward transport. Here, we explore a combination of remote sensing products and an exceptional latitudinal dataset of plankton counts collected in the surfzone and offshore in OR and WA coastal waters. Records of satellite ocean color for this period support the new finding that blooms were concurrent in OR and WA waters, with no evidence for latitudinal propagation as had been previously suggested. Plankton analyses further indicate that there was a rapid and synchronized increase of A. sanguinea between late August and mid-September of 2009 along wide swaths of the OR and WA coasts. Bloom onset occurred during a prolonged quiescent and warm period in late August–early September, near the end of the March–October upwelling phase. An upwelling event in October likely contributed to foam production through vertical mixing of A. sanguinea rich waters. Bloom intensity peaked earlier and at higher levels in WA waters as compared to OR with cell concentrations exceeding 1.5×106cellsL−1 (WA) and ∼350,000cellsL−1 (OR). In OR samples, A. sanguinea cells comprised upwards of 90% of dinoflagellate cell counts and ∼30% of total phytoplankton cells. At some locations, A. sanguinea persisted well into November–December of 2009, during which time satellite sea surface temperature records indicated anomalously warm surface waters (up to ∼5°C greater than climatological means). Taken together, the data reveal a HAB event of a magnitude unprecedented in over a decade of observations. We hypothesize that these blooms originated from either a cryptic cyst bed and/or a pelagic seed bank of viable vegetative cells.

Ribosomal DNA Organization Patterns within the Dinoflagellate Genus Alexandrium as Revealed by FISH: Life Cycle and Evolutionary Implications

Dinoflagellates are a group of protists whose genome differs from that of other eukaryotes in terms of size (contains up to 250pg per haploid cell), base composition, chromosomal organization, and gene expression. But rDNA gene mapping of the active nucleolus in this unusual eukaryotic genome has not been carried out thus far. Here we used FISH in dinoflagellate species belonging to the genus Alexandrium (genome sizes ranging from 21 to 170 pg of DNA per haploid genome) to localize the sequences encoding the 18S, 5.8S, and 28S rRNA genes. The results can be summarized as follows: 1) Each dinoflagellate cell contains only one active nucleolus, with no hybridization signals outside it. However, the rDNA organization varies among species, from repetitive clusters forming discrete nuclear organizer regions (NORs) in some to specialized "ribosomal chromosomes" in other species. The latter chromosomes, never reported before in other eukaryotes, are mainly formed by rDNA genes and appeared in the species with the highest DNA content. 2) Dinoflagellate chromosomes are first characterized by several eukaryotic features, such as structural differentiation (centromere-like constrictions), size differences (dot chromosomes), and SAT (satellite) chromosomes. 3) NOR patterns prove to be useful in discriminating between cryptic species and life cycle stages in protists.

Cold-Induced Cysts of the Photosynthetic DinoflagellateLingulodinium polyedrumHave an Arrested Circadian Bioluminescence Rhythm and Lower Levels of Protein Phosphorylation

Dinoflagellates are microscopic, eukaryotic, and primarily marine plankton. Temporary cyst formation is a well-known physiological response of dinoflagellate cells to environmental stresses. However, the molecular underpinnings of cold-induced cyst physiology have never been described. Cultures of the photosynthetic dinoflagellate Lingulodinium polyedrum readily form temporary cysts when placed at low (8°C±1°C) temperature and excyst to form normal motile cells following a return to normal temperature (18°C±1°C). The normal circadian bioluminescence rhythm and the expected changes in Luciferin Binding Protein abundance were arrested in L. polyedrum cysts. Furthermore, after excystment, the bioluminescence rhythm initiates at a time corresponding to zeitgeber 12, independent of the time when the cells encysted. Phosphoprotein staining after two-dimensional polyacrylamide gel electrophoresis, as well as column-based phosphoprotein enrichment followed by liquid chromatography tandem mass spectrometry, showed cyst proteins are hypophosphorylated when compared with those from motile cells, with the most marked decreases found for predicted Casein Kinase2 target sites. In contrast to the phosphoproteome, the cyst proteome is not markedly different from motile cells, as assessed by two-dimensional polyacrylamide gel electrophoresis. In addition to changes in the phosphoproteome, RNA sequencing revealed that cysts show a significant decrease in the levels of 132 RNAs. Of the 42 RNAs that were identified by sequence analysis, 21 correspond to plastid-encoded gene products and 11 to nuclear-encoded cell wall/plasma membrane components. Our data are consistent with a model in which the highly reduced metabolism in cysts is achieved primarily by alterations in the phosphoproteome. The stalling of the circadian rhythm suggests temporary cysts may provide an interesting model to address the circadian system of dinoflagellates.

A Quantitative Assessment of the Role of the Parasite Amoebophrya in the Termination of Alexandrium fundyense Blooms within a Small Coastal Embayment

Parasitic dinoflagellates of the genus Amoebophrya infect free-living dinoflagellates, some of which can cause harmful algal blooms (HABs). High prevalence of Amoebophrya spp. has been linked to the decline of some HABs in marine systems. The objective of this study was to evaluate the impact of Amoebophrya spp. on the dynamics of dinoflagellate blooms in Salt Pond (MA, USA), particularly the harmful species Alexandrium fundyense. The abundance of Amoebophrya life stages was estimated 3–7 days per week through the full duration of an annual A. fundyense bloom using fluorescence in situ hybridization coupled with tyramide signal amplification (FISH- TSA). More than 20 potential hosts were recorded including Dinophysis spp., Protoperidinium spp. and Gonyaulax spp., but the only dinoflagellate cells infected by Amoebophrya spp. during the sampling period were A. fundyense. Maximum A. fundyense concentration co-occurred with an increase of infected hosts, followed by a massive release of Amoebophrya dinospores in the water column. On average, Amoebophrya spp. infected and killed ∼30% of the A. fundyense population per day in the end phase of the bloom. The decline of the host A. fundyense population coincided with a dramatic life-cycle transition from vegetative division to sexual fusion. This transition occurred after maximum infected host concentrations and before peak infection percentages were observed, suggesting that most A. fundyense escaped parasite infection through sexual fusion. The results of this work highlight the importance of high frequency sampling of both parasite and host populations to accurately assess the impact of parasites on natural plankton assemblages.

Physical–biological coupling induced aggregation mechanism for the formation of high biomass red tides in low nutrient waters

Port Shelter is a semi-enclosed bay in northeast Hong Kong where high biomass red tides are observed to occur frequently in narrow bands along the local bathymetric isobars. Previous study showed that nutrients in the Bay are not high enough to support high biomass red tides. The hypothesis is that physical aggregation and vertical migration of dinoflagellates appear to be the driving mechanism to promote the formation of red tides in this area. To test this hypothesis, we used a high-resolution estuarine circulation model to simulate the near-shore water dynamics based on in situ measured temperature/salinity profiles, winds and tidal constitutes taken from a well-validated regional tidal model. The model results demonstrated that water convergence occurs in a narrow band along the west shore of Port Shelter under a combined effect of stratified tidal current and easterly or northeasterly wind. Using particles as dinoflagellate cells and giving diel vertical migration, the model results showed that the particles aggregate along the convergent zone. By tracking particles in the model predicted current field, we estimated that the physical–biological coupled processes induced aggregation of the particles could cause 20–45 times enhanced cell density in the convergent zone. This indicated that a high cell density red tide under these processes could be initialized without very high nutrients concentrations. This may explain why Port Shelter, a nutrient-poor Bay, is the hot spot for high biomass red tides in Hong Kong in the past 25 years. Our study explains why red tide occurrences are episodic events and shows the importance of taking the physical–biological aggregation mechanism into consideration in the projection of red tides for coastal management.

Genomics Study of the Exposure Effect of Gymnodinium catenatum, a Paralyzing Toxin Producer, on Crassostrea gigas' Defense System and Detoxification Genes

Background Crassostrea gigas accumulates paralytic shellfish toxins (PST) associated with red tide species as Gymnodinium catenatum. Previous studies demonstrated bivalves show variable feeding responses to toxic algae at physiological level; recently, only one study has reported biochemical changes in the transcript level of the genes involved in C. gigas stress response.Principal FindingsWe found that 24 h feeding on toxic dinoflagellate cells (acute exposure) induced a significant decrease in clearance rate and expression level changes of the genes involved in antioxidant defense (copper/zinc superoxide dismutase, Cu/Zn-SOD), cell detoxification (glutathione S-transferase, GST and cytochrome P450, CPY450), intermediate immune response activation (lipopolysaccharide and beta glucan binding protein, LGBP), and stress responses (glutamine synthetase, GS) in Pacific oysters compared to the effects with the non-toxic microalga Isochrysis galbana. A sub-chronic exposure feeding on toxic dinoflagellate cells for seven and fourteen days (30×103 cells mL−1) showed higher gene expression levels. A significant increase was observed in Cu/Zn-SOD, GST, and LGBP at day 7 and a major increase in GS and CPY450 at day 14. We also observed that oysters fed only with G. catenatum (3×103 cells mL−1) produced a significant increase on the transcription level than in a mixed diet (3×103 cells mL−1 of G. catenatum+0.75×106 cells mL−1 I. galbana) in all the analyzed genes.ConclusionsOur results provide gene expression data of PST producer dinoflagellate G. catenatum toxic effects on C. gigas, a commercially important bivalve. Over expressed genes indicate the activation of a potent protective mechanism, whose response depends on both cell concentration and exposure time against these toxic microalgae. Given the importance of dinoflagellate blooms in coastal environments, these results provide a more comprehensive overview of how oysters respond to stress generated by toxic dinoflagellate exposure.

Effective growth of dinoflagellate Prorocentrum minimum by cultivating the cells using municipal wastewater as nutrient source

Several studies have been conducted worldwide in order to develop a more economical method for mass algal cultivation so that more cost-effective biomass production can be accessed. One of the directions is to reduce production costs by using wastewater as a nutrient source in algal cell cultivation. The growth ability of a red-tide causative dinoflagellate species, Prorocentrum minimum, in various concentrations of local urban wastewater was examined in this study. The highest exponential growth rate and maximum cell density (MCD) were achieved when autoclaved 10% wastewater was used for cell cultivation, although the cells could survive in 0.01-100% wastewater. Both growth rate and MCD of the cells in wastewater were found to be substantially higher than that in optimized L1 culture medium. This research highlights the potential of using wastewater as a cost-effective approach for mass cultivation of dinoflagellate cells with consequent production of valuable microalgal biomass.

Bathyphotometer bioluminescence potential measurements: A framework for characterizing flow agitators and predicting flow-stimulated bioluminescence intensity

Bathyphotometer measurements of bioluminescence are used as a proxy for the abundance of luminescent organisms for studying population dynamics; the interaction of luminescent organisms with physical, chemical, and biological oceanographic processes; and spatial complexity especially in coastal areas. However, the usefulness of bioluminescence measurements has been limited by the inability to compare results from different bathyphotometer designs, or even the same bathyphotometer operating at different volume flow rates. The primary objective of this study was to compare measurements of stimulated bioluminescence of four species of cultured dinoflagellates, the most common source of bioluminescence in coastal waters, using two different bathyphotometer flow agitators as a function of bathyphotometer volume flow rate and dinoflagellate concentration. For both the NOSC and BIOLITE flow agitators and each species of dinoflagellate tested, there was a critical volume flow rate, above which average bioluminescence intensity, designated as bathyphotometer bioluminescence potential (BBP), remained relatively constant and scaled directly with dinoflagellate cell concentration. At supra-critical volume flow rates, the ratio of BIOLITE to NOSC BBP was nearly constant for the same species studied, but varied between species. The spatial pattern and residence time of flash trajectories within the NOSC flow agitator indicated the presence of dominant secondary recirculating flows, where most of the bioluminescence was detected. A secondary objective (appearing in the Appendix) was to study the feasibility of using NOSC BBP to scale flow-stimulated bioluminescence intensity across similar flow fields, where the contributing composition of luminescent species remained the same. Fully developed turbulent pipe flow was chosen because it is hydrodynamically well characterized. Average bioluminescence intensity in a 2.54-cm i.d. pipe was highly correlated with wall shear stress and BBP. This correlation, when further scaled by pipe diameter, effectively predicted bioluminescence intensity in fully developed turbulent flow in a 0.83-cm i.d. pipe. Determining similar correlations between other bathyphotometer flow agitators and flow fields will allow bioluminescence potential measurements to become a more powerful tool for the oceanographic community.

Germination of phytoplankton resting cells from surface sediments in two areas of the Southern Chinese coastal waters

AbstractTo understand the role of phytoplankton resting cells in the outbreak of algal blooms, particularly harmful algal blooms, surface sediments were collected monthly from April 2007 to March 2008 from two bays near the international ports in the Southern Chinese coastal waters. Sieved sediments were incubated for 20 and 40 days, and germinated vegetative cells were observed. Altogether, 97 taxa were recorded, of which 50 were diatoms and 35 dinoflagellates. Vegetative cells of cyanobacteria, chlorophytes, dictyophytes, euglenophytes, haptophytes, and raphidophytes were also observed. Centric diatoms such as Chaetoceros, Melosira, Skeletonema, and Thalassiosira dominated. Scrippsiella, Gymnodinium, and Alexandrium were common dinoflagellate taxa. Diatom spores germinated in samples from all seasons but were abundant in the autumn and winter samples. Low numbers of dinoflagellate cells germinated in the winter samples. The nanophytoplankton taxa, Gymnodinium corii and Chrysochromulina sp., which have not been recorded in the previous phytoplankton surveys, were abundant, suggesting either their new appearance in the water column or perhaps that they were overlooked in routine phytoplankton monitoring due to their small sizes. Vegetative cells of harmful or potentially harmful taxa were germinated, and some of them such as Amphidinium, Gambierdiscus, Ostreopsis, and Coolia have not previously been reported in the study area. Based on the results of the incubation of sediments from the two bays near the international ports, it is suggested that international shipping increases the risk of the introduction of new phytoplankton species and thus promotes the incidence of harmful algal blooms.

Life Cycle of the pseudocolonial dinoflagellate Polykrikos kofoidii (Gymnodiniales, Dinoflagellata).

The athecate, pseudocolonial polykrikoid dinoflag-ellates show a greater morphological complexity than many other dinoflagellate cells and contain not only elaborate extrusomes but sulci, cinguli, flagellar pairs, and nuclei in multiple copies. Among polykrikoids, Polykrikos kofoidii is a common species that plays an important role as a grazer of toxic planktonic algae but whose life cycle is poorly known. In this study, the main life cycle stages of P. kofoidii were examined and documented for the first time. The formation of gametes, 2-zooid-1-nucleus stages very different from vegetative cells, was observed and the process of gamete fusion, isogamy, was recorded. Karyogamy followed shortly after completed plasmogamy. A complex reorganization of furrows (cinguli and sulci) and flagella followed zygote formation, resulting in a 4-zooid zygote with one nucleus. The fate of zygotes under different nutritional conditions was also investigated; well-fed zygotes were able to reenter the vegetative cycle via meiotic divisions as indicated by nuclear cyclosis. However, nuclear cyclosis was preceded by a presumably mitotic division of the primary zygote nucleus which by definition would imply that P. kofoidii has a diplohaplontic life cycle. Nuclear cyclosis in germlings hatched from spiny resting cysts indicate that these cysts are of zygote origin (hypnozygotes). Hypnozygote formation, cyst hatching, the morphology of the germling (a 1-zooid cell), and its development into a normal pseudocolony are documented here for the first time. There is evidence that P. kofoidii has a system of complex heterothallism.

Spatio-temporal variation of microphytoplankton in the upwelling system of the south-eastern Arabian Sea during the summer monsoon of 2009

The phytoplankton standing crop was assessed in detail along the South Eastern Arabian Sea (SEAS) during the different phases of coastal upwelling in 2009. During phase 1 intense upwelling was observed along the southern transects (8°N and 8.5°N). The maximum chlorophyll a concentration (22.7mgm −3) was observed in the coastal waters off Thiruvananthapuram (8.5°N). Further north there was no signature of upwelling, with extensive Trichodesmium erythraeum blooms. Diatoms dominated in these upwelling regions with the centric diatom Chaetoceros curvisetus being the dominant species along the 8°N transect. Along the 8.5°N transect pennate diatoms like Nitzschia seriata and Pseudo-nitzschia sp. dominated. During phase 2, upwelling of varying intensity was observed throughout the study area with maximum chlorophyll a concentrations along the 9°N transect (25mgm−3) with Chaetoceros curvisetus as the dominant phytoplankton. Along the 8.5°N transect pennate diatoms during phase 1 were replaced by centric diatoms like Chaetoceros sp. The presence of solitary pennate diatoms Amphora sp. and Navicula sp. were significant in the waters off Kochi. Upwelling was waning during phase 3 and was confined to the coastal waters of the southern transects with the highest chlorophyll a concentration of 11.2mgm−3. Along with diatoms, dinoflagellate cell densities increased in phases 2 and 3. In the northern transects (9°N and 10°N) the proportion of dinoflagellates was comparatively higher and was represented mainly by Protoperidinium spp., Ceratium spp. and Dinophysis spp.

Spatial variation of phytoplankton assemblages and biomass in the New Zealand sector of the Southern Ocean during the late austral summer 2008

Up until now, only the floristic composition of large phytoplankton species has been studied in the Southern Ocean immediately south of New Zealand. To fill the gaps in our knowledge of the Antarctic phytoplankton flora and biomass, in relation to physical and chemical environments, New Zealand’s IPY project sampled water between New Zealand and Antarctica and into the Ross Sea in late summer. Near surface phytoplankton samples, collected between New Zealand and Antarctica, and vertical stations, between Scott Island and the Ross Sea ice shelf, allowed us to contrast the flora and biomass in the Ross Sea/Antarctic and Subantarctic waters. Spatially, diatoms were found to be more diverse in both shelf and offshore Antarctic than in Subantarctic waters. Over the Ross Sea shelf, the pennate diatom, Fragilariopsis kerguelensis, and a colonial prymnesiophyte, Phaeocystis antarctica, dominated both numerically and in biomass, contributing to greatest levels of chlorophyll a and cell carbon biomass over the shelf. In both shelf and Antarctic waters, heterotrophic taxa accounted for a greater proportion of total dinoflagellate cell carbon (avg. 95.4 and 91.9 %, respectively) than autotrophic, while in the Subantarctic waters, the reverse was true (avg. 48.6 %). A small number of potentially harmful dinoflagellates, Karenia and Karlodinium spp., were found for the first time in the New Zealand sector of the Southern Ocean. This study represents the first thorough description of the spatial variation of the floristic composition and biomass of phytoplankton in relation to physical and chemical properties, from New Zealand to Antarctica.

Toxin production, retention, and extracellular release by Dinophysis acuminata during extended stationary phase and culture decline

Dinophysis spp. produce diarrhetic shellfish poisoning (DSP) toxins and pectenotoxins. The extent to which the dinoflagellate cells retain their toxicity in stationary phase, a period when cells are most toxic, and their transition into cell death is not known. Here we present results on the production, recycling, retention, and release of toxins from a monoculture of Dinophysis acuminata during these two important stages. Once stationary phase was reached, cultures were divided between light and dark treatments to identify if light influenced toxin dynamics. Light was required for long-term cell maintenance (>2 months) of D. acuminata in the absence of prey, however, in the dark, cells in stationary phase survived on reserves alone for four weeks before beginning to decline. Cells maintained relatively constant levels of intracellular OA (0.39±0.03pg/cell, 0.44±0.05pg/cell), DTX1 (0.45±0.09pg/cell, 0.64±0.10pg/cell) and PTX2 (10.4±1.4pg/cell, 11.0±1.9pg/cell) in the dark and light treatments, respectively, throughout stationary phase and into culture decline. Toxin production was only apparent during late exponential and early stationary growth when cells were actively dividing. In general, the concentration of dissolved (extracellular) toxin in the medium significantly increased upon culture aging and decline; cells did not appear to be actively or passively releasing toxin during stationary phase, but rather extracellular release was likely a result of cell death. Light availability did not have an apparent effect on toxin production, quotas, or intracellular vs. extracellular distribution. Together these results suggest that a bloom of D. acuminata would retain its cellular toxicity or potency as long as the population is viable, and that cells under conditions of low light (e.g., at the boundary or below euphotic zone) and/or minimal prey could maintain toxicity for extended periods.