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The toxic dinoflagellate Karenia mikimotoi has been well-known for causing large-scale and dense harmful algal blooms (HABs) in coastal waters worldwide and serious economic loss in aquaculture and fisheries and other adverse effects on marine ecosystems. Whether K. mikimotoi forms resting cysts has been a puzzling issue regarding to the mechanisms of bloom initiation and geographic expansion of this species. We provide morphological and molecular confirmation of sexually produced thin-walled resting cysts by K. mikimotoi based on observations of laboratory cultures and their direct detection in marine sediments. Light and scanning electron microscopy evidences for sexual reproduction include attraction and pairing of gametes, gamete fusion, formation of planozygote and thin-walled cyst, and the documentation of the thin-walled cyst germination processes. Evidence for cysts in marine sediments was in three aspects: positive PCR detection of cysts using species-specific primers in the DNA extracted from whole sediments; fluorescence in situ hybridization detection of cysts using FISH probes; and single-cell PCR sequencing for cysts positively labeled with FISH probes. The existence of sexually produced, thin-walled resting cysts by K. mikimotoi provides a possible mechanism accounting for the initiation of annually recurring blooms at certain regions and global expansion of the species during the past decades.

High-biomass blooms of the toxic dinoflagellate Pyrodinium bahamense occur most summers in Tampa Bay, Florida, USA, posing a recurring threat to ecosystem health. Like many dinoflagellates, P. bahamense forms immobile resting cysts that can be deposited on the seafloor-creating a seed bank that can retain the organism within the ecosystem and initiate future blooms when cysts germinate. In this study, we examined changes in the dormancy status of cysts collected from Tampa Bay and applied lessons from plant ecology to explore dormancy controls. Pyrodinium bahamense cysts incubated immediately after field collection displayed a seasonal pattern in dormancy and germination that matched the pattern of cell abundance in the water column. Newly deposited (surface) cysts and older (buried) cysts exhibited similar germination patterns, suggesting that a common mechanism regulates dormancy expression in new and mature cysts. Extended cool- and warm-temperature conditioning of field-collected cysts altered the cycle of dormancy compared with that of cysts in nature, with the duration of cool temperature exposure being the best predictor of when cysts emerged from dormancy. Extended warm conditioning, on the other hand, elicited a return to dormancy, or secondary dormancy, in nondormant cysts. These results directly demonstrate environmental induction of secondary dormancy in dinoflagellates-a mechanism common and thoroughly documented in higher plants with seasonal growth cycles. Our findings support the hypothesis that a seasonal cycle in cyst germination drives P. bahamense bloom periodicity in Tampa Bay and point to environmentally induced secondary dormancy as an important regulatory factor of that cycle.

Diatoms are one of the dominant groups in phytoplankton communities of the western Antarctic Peninsula (WAP). Although generally well-studied, little is known about size dependent photophysiological responses in diatom bloom formation and succession. To increase this understanding, four Antarctic diatom species covering two orders of magnitude in cell size were isolated in northern Marguerite Bay (WAP). Fragilariopsis sp., Pseudo-nitzschia cf. subcurvata, Thalassiosira cf. antarctica, and Proboscia cf. alata were acclimated to three different irradiances after which photophysiology, electron transport, carbon fixation and growth were assessed. The small species Fragilariopsis sp., Pseudo-nitzschia cf. subcurvata, and large species Proboscia cf. alata showed similar photoacclimation to higher irradiances with a decrease in cellular chlorophyll a and an increase in chlorophyll a specific absorption and xanthophyll cycle pigments and activity. In contrast, pigment concentrations and absorption remained unaffected by higher irradiances in the large species Thalassiosira cf. antarctica. Overall, the small species showed significantly higher growth rates compared to the large species, which was related to relatively high light harvesting capacity and electron transport rates in the smaller species. However, photophysiological responses related to photoinhibition and photoprotection and carbon fixation showed no relationship to cell size. The present study supports the dominance of small diatoms at low irradiances during winter and early spring, but does not provide photophysiological evidence for the dominance of large diatoms during the phytoplankton bloom in the WAP. This suggests that other factors such as grazing and nutrient availability are likely to play a major role in diatom bloom formation. This article is protected by copyright. All rights reserved.

The bloom-forming freshwater alga Gonyostomum semen is associated with acidic, mesotrophic brown water lakes in boreal regions. However, researchers have been unable to conclusively link G. semen abundance and bloom formation to typical brown water lake traits, that is, high water color and DOC (dissolved organic carbon) concentrations. Iron is a main driver of water color in boreal lakes, and a recent study of lake monitoring data indicated a connection between lakes with high G. semen abundance and iron concentrations >200 µg · L−1. Thus, iron may be the missing link in explaining G. semen abundance and growth dynamics. We experimentally assessed the effects of different iron concentrations above or below 200 µg · L−1 on the growth of G. semen batch monocultures. Iron concentrations 200 µg · L−1 did not. Moreover, the iron concentration of the medium required for growth was higher than for other common phytoplankton (Microcystis botrys and Chlamydomonas sp.) included in the experiment. These results indicate that G. semen requires high levels of iron in the lake environment. Consequently, this and previous findings using lake monitoring data support the hypothesis that high concentrations of iron favor the formation of high-density G. semen blooms in boreal brown water lakes. As lakes get browner in a changing climate, monitoring iron levels could be a potential tool to identify lakes at risk for G. semen blooms, especially among lakes that provide ecosystem services to society.

The poorly understood filamentous cyanobacterium Pseudanabaena is commonly epiphytic on Microcystis colonies and their abundances are often highly correlated during blooms. The response and adaptation of Microcystis to iron limitation have been extensively studied, but the strategies Pseudanabaena uses to respond to iron limitation are largely unknown. Here, physiological responses to iron limitation were compared between one Pseudanabaena and two Microcystis strains grown under different light intensities. The results showed that low-intensity light exacerbated, but high-intensity light alleviated, the negative effect of iron limitation on Pseudanabaena growth relative to two Microcystis strains. It was found that robust light-harvesting and photosynthetic efficiency allowed adaptation of Pseudanabaena to low light availability relative to two Microcystis strains only during iron sufficiency. The results also indicated that a larger investment in the photosynthetic antenna probably contributed to light/iron co-limitation of Pseudanabaena relative to two Microcystis strains under both light and iron limitation. Furthermore, the lower antenna pigments/chlorophyll a ratio and photosynthetic efficiency, and higher nonphotochemical quenching and saturation irradiance provided Pseudanabaena photoadaptation and photoprotection advantages over the two Microcystis strains under the high-light condition. The lower investment in antenna pigments of Pseudanabaena than the two Microcystis strains under high-light intensity is likely an efficient strategy for both saving iron quotas and decreasing photosensitivity. Therefore, when compared with Microcystis, the high plasticity of antenna pigments, along with the excellent photoadaptation and photoprotection ability of Pseudanabaena, probably ensures its ecological success under iron limitation when light is sufficient.

Nitrogen fixation has been proposed as a mechanism that allows the diazotrophic cyanobacterium, Cylindrospermopsis raciborskii, to bloom in nitrogen-limited freshwater systems. However, it is unclear whether dinitrogen fixation (N2 fixation) can supplement available dissolved inorganic nitrogen (DIN) for growth, or only provides minimum nitrogen (N) for cell maintenance under DIN deplete conditions. Additionally, the rate at which cells can switch between DIN use and N2 fixation is unknown. This study investigated N2 fixation under a range of nitrate concentrations. Cultures were grown with pretreatments of nitrate replete (single dose 941 μmol NO3- · L-1 ) and N-free conditions and then either received a single dose of 941 μmol NO3- · L-1 (N941), 118 μmol NO3- · L-1 (N118) or 0 N. Heterocysts appeared from days 3 to 5 when treatments of high NO3- were transferred to N free media (N941:N0), and from day 5 in N941 transferred to N118 treatments. Conversely, transferring cells from N0 to N941 resulted in heterocysts being discarded from day 3 and day 5 for N0:N118. Heterocyst appearance correlated with a detectable rate of N2 fixation and up-regulation of nifH gene expression, the discard of heterocysts occurred after sequential reduction of nifH expression and N2 fixation. Nitrate uptake rates were not affected by pretreatment, suggesting no regulation or saturation of this uptake pathway. These data demonstrate that for C. raciborskii, N2 fixation is regulated by the production or discard of heterocysts. In conclusion, this study has shown that N2 fixation only provides enough N to support relatively low growth under N-limited conditions, and does not supplement available nitrate to increase growth rates.

International audience; We used a multistrain approach to study the intra‐ and interspecific variability of the growth rates of three Pseudo‐nitzschia species – P. australis, P. fraudulenta, and P. pungens – and of their domoic acid (DA) production. We carried out mating and batch experiments to investigate the respective effects of strain age and cell size, and thus the influence of their life cycle on the physiology of these species. The cell size – life cycle relationship was characteristic of each species. The influence of age and cell size on the intraspecific variability of growth rates suggests that these characteristics should be considered cautiously for the strains used in physiological studies on Pseudo‐nitzschia species. The results from all three species do not support the hypothesis of a decrease in DA production with time since isolation from natural populations. In P. australis, the cellular DA content was rather a function of cell size. More particularly, cells at the gametangia stage of their life cycle contained up to six times more DA than smaller or larger cells incapable of sexual reproduction. These findings reveal a link between P. australis life cycle and cell toxicity. This suggest that life cycle dynamics in Pseudo‐nitzschia natural populations may influence bloom toxicity.

A recent rise in the relative abundance of Discostella species (D. stelligera and D. pseudostelligera) has been well documented from sedimentary diatom assemblages across the Northern Hemisphere. This unprecedented change over the last ~150 years has been linked to rises in atmospheric temperatures, changes in ice cover, and/or increases in thermal stability, among other factors. The bi-weekly monitoring data from two boreal lakes at the Experimental Lakes Area (ELA) in northwestern Ontario were analyzed across seasons (spring, summer, and fall) and decades (1970s-2000s). We found that Discostella species are primarily spring/early summer bloomers (i.e., late April to June) in these lakes and changes in concentrations of Discostella over time were most pronounced in the spring or early summer months. Increases in Discostella abundance over time may be linked to earlier ice-off and a longer period of spring turnover, resulting from increased winter and spring temperatures. It is also possible that a trophic mismatch between the spring diatom bloom and zooplankton is occurring, thus reducing diatom loss rates, and resulting in greater overall abundance. Moreover, the spring dominance of Discostella in our study lakes occurred at a time of the year when nutrient concentrations were at their highest seasonally, suggesting that these taxa are neither limited directly by nutrients, nor responding to enhanced stratification during the summer months in these lakes.

The abundance of Aulacoseira granulata (Ehrenburg) Simonsen and Gloeocystis planctonica (West & G.S.West) Lemmermann was assessed during the summers of 2005 and 2010 in the eutrophic Fox River, Wisconsin, USA. In both years, a mid-summer bloom of G. planctonica was followed by the rapid growth of A. granulata. Laboratory experiments in which A. granulata was grown in cell-free filtrate of a G. planctonica culture revealed that the growth of A. granulata was stimulated in the G. planctonica-treated medium relative to controls. This effect was detected when dormant A. granulata cells were used as the source culture for the experiment but not when actively growing cells were used. Dormant A. granulata also grew more rapidly in river water collected after the 2010 G. planctonica bloom relative to river water collected before the bloom. These results suggest that the summer bloom of A. granulata in the river was stimulated by G. planctonica. This relationship can be described as stimulated rejuvenation, an interaction where the transition of an algal resting stage into active growth is triggered by exposure to another species.

Algal blooms are a worldwide phenomenon and the biological interactions that underlie their regulation are only just beginning to be understood. It is established that algal microorganisms associate with many other ubiquitous, oceanic organisms, but the interactions that lead to the dynamics of bloom formation are currently unknown. To address this gap, we used network approaches to investigate the association patterns among microeukaryotes and bacterioplankton in response to a natural Scrippsiella trochoidea bloom. This is the first study to apply network approaches to bloom dynamics. To this end, terminal restriction fragment (T-RF) length polymorphism analysis showed dramatic changes in community compositions of microeukaryotes and bacterioplankton over the blooming period. A variance ratio test revealed significant positive overall associations both within and between microeukaryotic and bacterioplankton communities. An association network generated from significant correlations between T-RFs revealed that S. trochoidea had few connections to other microeukaryotes and bacterioplankton and was placed on the edge. This lack of connectivity allowed for the S. trochoidea sub-network to break off from the overall network. These results allowed us to propose a conceptual model for explaining how changes in microbial associations regulate the dynamics of an algal bloom. In addition, key T-RFs were screened by principal components analysis, correlation coefficients, and network analysis. Dominant T-RFs were then identified through 18S and 16S rRNA gene clone libraries. Results showed that microeukaryotes clustered predominantly with Dinophyceae and Perkinsea while the majority of bacterioplankton identified were Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes. The ecologi-cal roles of both were discussed in the context of these findings.

Iron availability limits primary productivity in large areas of the world's oceans. Ascertaining the iron status of phytoplankton is essential for understanding the factors regulating their growth and ecology. We developed an incubation-independent, molecular-based approach to assess the iron nutritional status of specific members of the diatom community, initially focusing on the ecologically important pennate diatom Pseudo-nitzschia. Through a comparative transcriptomic approach, we identified two genes that track the iron status of Pseudo-nitzschia with high fidelity. The first gene, ferritin (FTN), encodes for the highly specialized iron storage protein induced under iron-replete conditions. The second gene, ISIP2a, encodes an iron-concentrating protein induced under iron-limiting conditions. In the oceanic diatom Pseudo-nitzschia granii (Hasle) Hasle, transcript abundance of these genes directly relates to changes in iron availability, with increased FTN transcript abundance under iron-replete conditions and increased ISIP2a transcript abundance under iron-limiting conditions. The resulting ISIP2a:FTN transcript ratio reflects the iron status of cells, where a high ratio indicates iron limitation. Field samples collected from iron grow-out microcosm experiments conducted in low iron waters of the Gulf of Alaska and variable iron waters in the California upwelling zone verify the validity of our proposed Pseudo-nitzschia Iron Limitation Index, which can be used to ascertain in situ iron status and further developed for other ecologically important diatoms.

Since the 1970s, Puget Sound, Washington State, USA, has experienced an increase in detections of paralytic shellfish toxins (PSTs) in shellfish due to blooms of the harmful dinoflagellate Alexandrium. Natural patterns of climate variability, such as the Pacific Decadal Oscillation (PDO), and changes in local environmental factors, such as sea surface temperature (SST) and air temperature, have been linked to the observed increase in PSTs. However, the lack of observations of PSTs in shellfish prior to the 1950s has inhibited statistical assessments of longer-term trends in climate and environmental conditions on Alexandrium blooms. After a bloom, Alexandrium cells can enter a dormant cyst stage, which settles on the seafloor and then becomes entrained into the sedimentary record. In this study, we created a record of Alexandrium spp. cysts from a sediment core obtained from Sequim Bay, Puget Sound. Cyst abundances ranged from 0 to 400 cysts · cm(-3) and were detected down-core to a depth of 100 cm, indicating that Alexandrium has been present in Sequim Bay since at least the late 1800s. The cyst record allowed us to statistically examine relationships with available environmental parameters over the past century. Local air temperature and sea surface temperature were positively and significantly correlated with cyst abundances from the late 1800s to 2005; no significant relationship was found between PDO and cyst abundances. This finding suggests that local environmental variations more strongly influence Alexandrium population dynamics in Puget Sound when compared to large-scale changes.

Harmful blooms formed by species of the dinoflagellate Cochlodinium have caused massive fish kills and substantial economic losses in the Pacific Ocean. Recently, prominent blooms of Cochlodinium have occurred in central and southern California (2004-2008), and Cochlodinium cells are now routinely observed in microscopical analysis of algal assemblages from Californian coastal waters. The first documented economic loss due to a Cochlodinium bloom in California occurred in Monterey Bay and resulted in the mortality of commercially farmed abalone. Increasing occurrences of Cochlodinium blooms, the fact that these cells preserve poorly using standard techniques, and the difficulty of identifying preserved specimens using morphological criteria make Cochlodinium species prime candidates for the development of a quantitative real-time polymerase chain reaction (qPCR) approach. The 18S rDNA gene sequenced from Cochlodinium cells obtained from California coastal waters, as well as GenBank sequences of Cochlodinium, were used to design and test a Molecular Beacon(®) approach. The qPCR method developed in this study is species specific, sensitive for the detection of C. fulvescens that has given rise to the recent blooms in the eastern Pacific Ocean, and spans a dynamic abundance range of seven orders of magnitude. Initial application of the method to archived field samples collected during blooms in Monterey Bay revealed no statistically significant correlations between gene copy number and environmental parameters. However, the onset of Cochlodinium blooms in central California was consistent with previously reported findings of correlations to decreased surface temperature and increased inputs of nitrogenous nutrients.

A giant form of Anadyomene, most similar to Anadyomene pavonina (J. Agardh) Wille, a rare and diminutive alga endemic to Florida, appeared as up to 10 m long net-like strands covering 10%-80% of a 0.5 km region of the 25-50 m deep Belizean outer reef slope where none had been present up to 12 months earlier. This new species, described herein as Anadyomene gigantodictyon Littler et D. S. Littler, is characterized by a unistratose blade or cluster of blades formed by the polychotomous branching of uniseriate veins, with the interstices, or spaces between the veins, completely or partially filled with cells that are smaller than those of the veins, with cylindrical to ovate cells. The cells at mid-blade are 1.7-2.0 mm in length and 0.2-0.3 mm diameter; interstitial cells are parallel and not juxtaposed. All cells are joined in one plane and form species-specific, fan-shaped patterns with secondary interstitial cells loosely or tightly woven.

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.

Blooms of the freshwater stalked diatom Didymosphenia geminata (Lyngb.) M. Schmidt in A. Schmidt typically occur in oligotrophic, unshaded streams and rivers. Observations that proliferations comprise primarily stalk material composed of extracellular polymeric substances (EPS) led us to ask whether or not the production of excessive EPS is favored under nutrient-limited, high-light conditions. We conducted experiments in outdoor flumes colonized by D. geminata using water from the oligotrophic, D. geminata-affected Waitaki River, South Island, New Zealand, to determine the relationship between D. geminata stalk length, cell division rates, and light intensity under ambient and nutrient-enriched conditions. Stalk lengths were measured in situ, and cell division rates were estimated as the frequency of dividing cells (FDC). FDC responded positively to increasing light intensity and to nutrient additions (N+P and P). Under ambient conditions, stalk length increased as light level increased except at low ambient light levels and temperature. Nutrient enrichment resulted in decreased stalk length and negative correlations with FDC, with this effect most evident under high light. Our results are consistent with the hypothesis that extensive stalk production in D. geminata occurs when cell division rates are nutrient limited and light levels are high. Thus, photosynthetically driven EPS production in the form of stalks, under nutrient-limited conditions, may explain the development of very high biomass in this species in oligotrophic rivers. The responses of FDC and stalk length under nutrient-replete conditions are also consistent with occurrences of D. geminata as a nondominant component of mixed periphyton communities in high-nutrient streams.

Cyanobacterial blooms are a common issue in eutrophic freshwaters, and some cyanobacteria produce toxins, threatening the health of humans and livestock. Microcystin, a representative cyanobacterial hepatotoxin, is frequently detected in most Korean lakes and reservoirs. This study developed predictive models for cyanobacterial bloom using artificial neural networks (ANNs; self-organizing map [SOM] and multilayer perceptron [MLP]), including an evaluation of related environmental factors. Fourteen environmental factors, as independent variables for predicting the cyanobacteria density, were measured weekly in the Daechung Reservoir from spring to autumn over 5 years (2001, 2003-2006). Cyanobacterial density was highly associated with environmental factors measured 3 weeks earlier. The SOM model was efficient in visualizing the relationships between cyanobacteria and environmental factors, and also for tracing temporal change patterns in the environmental condition of the reservoir. And the MLP model exhibited a good predictive power for the cyanobacterial density, based on the environmental factors of 3 weeks earlier. The water temperature and total dissolved nitrogen were the major determinants for cyanobacteria. The water temperature had a stronger influence on cyanobacterial growth than the nutrient concentrations in eutrophic waters. Contrary to general expectations, the nitrogen compounds played a more important role in bloom formation than the phosphorus compounds.

Sexual reproduction is documented for the first time in field populations of the pennate diatoms Pseudo-nitzschia australis Freng. and P. pungens (Grunow ex Cleve) Hasle (var. cingulata Villac and hybrids between var. cingulata and var. pungens). A bloom dominated by these species began on June 26, 2006, along Kalaloch Beach, Washington, USA, coincident with a drop in the Si(OH)4:NO3 ratio to below two. Multimodal size distributions were detected for both species, and synchronous auxosporulation occurred within the smallest size class during a 3-week window. Auxospores and initial cells created a new class of large cells, and cells in the intermediate size classes increased in abundance during auxosporulation. Mating cells of both species were attached to colonies of surf-zone diatoms. Paired gametangia, gametes, zygotes, auxospores, and large initial cells were found. Auxosporulation began first for P. pungens (June 30), apparently once a critical, high cell concentration was reached, followed by P. australis (July 5), when the total Pseudo-nitzschia cell concentration reached 929,000 cells · L−1. Low frequencies of auxosporulation occurred throughout the bloom but increased 4-fold for P. australis and 3-fold for P. pungens when macronutrients were reduced to low levels on July 11. A 2-year life cycle was estimated for P. australis and 3 years for P. pungens, both with annual auxosporulation. Domoic acid (DA) in razor clams reached a maximum of 38 μg DA · g−1 on July 18. A significant relationship existed between the percent of cells within the new size range and DA concentrations in razor clams on the same beach.

A naked dinoflagellate with a unique arrangement of chloroplasts in the center of the cell was isolated from the northern Baltic proper during a spring dinoflagellate bloom (March 2005). Morphological, ultrastructural, and molecular analyses revealed this dinoflagellate to be undescribed and belonging to the genus Gymnodinium F. Stein. Gymnodinium corollarium A. M. Sundstrom, Kremp et Daugbjerg sp. nov. possesses features typical of Gymnodinium sensu stricto, such as nuclear chambers and an apical groove running in a counterclockwise direction around the apex. Phylogenetic analyses based on partial nuclear-encoded LSU rDNA sequences place the species in close proximity to G. aureolum, but significant genetic distance, together with distinct morphological features, such as the position of chloroplasts, clearly justifies separation from this species. Temperature and salinity experiments revealed a preference of G. corollarium for low salinities and temperatures, confirming it to be a cold-water species well adapted to the brackish water conditions in the Baltic Sea. At nitrogen-deplete conditions, G. corollarium cultures produced small, slightly oval cysts resembling a previously unidentified cyst type commonly found in sediment trap samples collected from the northern and central open Baltic Sea. Based on LSU rDNA comparison, these cysts were assigned to G. corollarium. The cysts have been observed in many parts of the Baltic Sea, indicating the ecologic versatility of the species and its importance for the Baltic ecosystem.

The cosmopolitan bloom-forming diatom Skeletonema marinoi Sarno et Zingone is known to produce toxic polyunsaturated aldehydes (PUAs) in response to cell damage that can affect a diverse suite of organisms, including grazing species and competitor plankton species. The production of PUAs in nine different S. marinoi strains isolated at three different times of the year (spring, summer, and autumn) was assessed in relation to the predominant conditions at the time of isolation from Gullmar Fjord, Skagerrak. During the initial stages of growth, PUA production potential of S. marinoi was generally the highest in summer strains, although there was a substantial variation among strains isolated at the same time. Spring strains, however, showed a strong capacity for increased PUA production potential in later stage cultures with diminishing nutrient levels, reaching amounts similar to those observed in summer strains. In contrast, PUA production potentials of summer and autumn strains did not change significantly from the original values. There is negligible grazing pressure during the spring bloom in Gullmar Fjord, but a potential for high competition for resources, such as nutrients, toward the later stages of the bloom. In contrast, grazing pressure is much greater during summer and autumn, and there may also be nutrient limitation at this time. The PUA production potentials of S. marinoi appear to reflect the ecological conditions at the time of isolation with higher production potentials in strains isolated when conditions were likely to be less beneficial for survival.

Development of an Alexandrium minutum Halim bloom affecting a Mediterranean harbor was monitored in detail using a multidisciplinary approach. A. minutum was by far the most abundant species at and near the bloom maximum, but always coexisted with members of three additional dinoflagellate genera and prasinophytes. Bloom initiation (early February) occurred during prolonged influences of sunny weather conditions, when day length exceeded 10.5 h and water temperatures reached 10.2°C. Subsequent development toward its maximum (end of March) also relied on good weather conditions, with specific wind directions favoring accumulation of cells. Arrival of rainy weather, associated with frontal boundaries of large-scale low-atmospheric-pressure systems and characterized by reduced solar irradiance (heavy cloud coverage), opposite wind directions, and enhanced wind speeds, always caused temporal declines of the bloom. These declines were attributed to dispersal or displacement of algae, but a vertical migration of A. minutum cells toward the sediment was not excluded. Delayed inflows of excess terrestrial rainwater along the inner harbor wall strongly reduced salinity and prolonged a temporal decline far beyond influences of bad weather. The associated nutrient supply favored development of the phytoplankton population but reduced the toxin production of A. minutum cells. The HPLC-determined Gonyautoxin (GTX) 1 + 4/GTX 2 + 3 ratio strongly increased toward the bloom maximum. This ratio was influenced by nutrient status and cell density and has a potential value for monitoring developmental stages of blooms. Prolonged bad weather conditions eventually hindered continuation of bloom development, and subsequent declines of algal biomass were attributed to grazing.

Since 1998, a heterokont flagellate initially namedChattonella aff. verruculosa has formed recurrentextensive blooms in the North Sea and the Skager-rak, causing fish mortalities. Cells were isolatedfrom the 2001 bloom off the south coast of Norway,and monoalgal cultures were established and com-pared with the Chattonella verruculosa Y. Hara etChihara reference strain NIES 670 from Japan. Thecells in Norwegian cultured isolates were very vari-able in size and form, being large oblong (up to34 lm long) to small rounded (5–9 lm in diameter)with two unequal flagella, numerous chloroplasts,and mucocysts. The SSU and partial LSU rDNAsequences of strains from Norway and Japan werecompared and differed by 0.4% (SSU) and 1.3%(LSU), respectively. Five strains from Norway wereidentical in the LSU rDNA region. Phylogeneticanalyses based on heterokont SSU and concatenatedSSU + LSU rDNA sequences placed C. aff. verrucul-osa and the Japanese C. verruculosa within the cladeof Dictyochophyceae, with the picoflagellate Florenci-ella parvula Eikrem as the closest relative. Ultrastruc-ture, morphology, and pigment compositionsupported this affinity. We propose the nameVerrucophora farcimen sp. et gen. nov. for this flagellateand systematically place it within the class Dictyoch-ophyceae. Our studies also show that C. verruculosafrom Japan is genetically and morphologically dif-ferent but closely related to V. farcimen. The speciesis transferred from the class Raphidophyceae to theclass Dictyochophyceae and renamed Verrucophoraverruculosa. We propose a new order, Florenciellales,to accommodate V. farcimen, V. verruculosa, andF. parvula.Key index words: carotenoids; Chattonella; Dictyo-chophyceae;harmfulalgae;LSUrDNA;newtaxon;phylogeny; SSU rDNA; ultrastructureAbbreviations: G,gamma;GTR,general timereversible; I, proportion of invariable sites;MCMC, Markov chainMonteCarlo; ML, maximumlikelihoodIn April–May 1998, a heterokont phytoflagellateformed a massive bloom in the North Sea and theSkagerrak, off the coasts of Germany, Denmark,Sweden, and Norway, which killed 350 tons offarmed salmon as well as some feral fish (Aure et al.2001, Backe-Hansen et al. 2001). The responsibleorganism was initially identified as Chattonella aff.verruculosa because it resembled this speciesdescribed from Japan, but differed somewhat in cellsize, form, and growth pattern compared to previ-ous studies (Hara et al. 1994, Yamaguchi et al.1997). A bloom of a similar organism took place inApril–May 2000 in the German Bight and off theDanish Jutland coast (Lu and Go¨bel 2000). In 2001,C. aff. verruculosa again caused fish mortalities inNorway, and 1100 tons of reared salmon were killed.This time, the massive bloom developed in coldwaters (about 2 C–5 C) in the Kattegat and the

The bloom-forming cyanobacterium Microcystis aeruginosa (Kutz.) Kutz. 854 was cultured with 1.05 W · m⁻² ultraviolet-B radiation (UVBR) for 3 h every day, and the CO₂-concentrating mechanism (CCM) within this species as well as effects of UVBR on its operation were investigated. Microcystis aeruginosa 854 possessed at least three inorganic carbon transport systems and could utilize external HCO₃⁻ and CO₂ for its photosynthesis. The maximum photosynthetic rate was approximately the same, but the apparent affinity for dissolved inorganic carbon was significantly decreased from 74.7 μmol · L⁻¹ in the control to 34.7 μmol · L⁻¹ in UVBR-treated cells. At 150 μmol · L⁻¹ KHCO₃ and pH 8.0, Na⁺-dependent HCO₃⁻ transport contributed 43.4%-40.2% to the photosynthesis in the control and 34.5%-31.9% in UVBR-treated cells. However, the contribution of Na⁺-independent HCO₃⁻ transport increased from 8.7% in the control to 18.3% in UVBR-treated cells. The contribution of CO₂-uptake systems showed little difference: 47.9%-51.0% in the control and 49.8%-47.2% in UVBR-treated cells. Thus, the rate of total inorganic carbon uptake was only marginally affected, although UVBR had a differential effect on various inorganic carbon transporters. However, the number of carboxysomes in UVBR-treated cells was significantly decreased compared to that in the control.

The harmful bloom alga Aureococcus anophagefferens Hargraves et Sieburth can survive prolonged darkness, which could facilitate overwinter survival and dispersal by anthropogenic vectors such as ballast water. Experiments were conducted to examine the biochemical and photosynthetic changes in cells during dark storage. Cells were stored in the dark for periods from 2 to 14 days and were sampled at days 0, 1, 2, 4, 7, 10, and 14. Samples from days 2, 7, and 14 were monitored during a recovery period of 4–5 days. Physiological and photosynthetic parameters were measured during the dark storage and recovery periods. Cultures resumed growth quickly when returned to light, and bacterial counts remained constant during the dark storage period but increased rapidly during recovery periods. Cellular protein, carbohydrate, and lipid concentrations declined slightly during the dark period. There were no changes in chl a per cell or in RUBISCO per cell during 14 days of darkness. The data therefore suggest that A. anophagefferens is able to maintain its photosynthetic apparatus during dark storage periods of at least 2 weeks and relies on cellular reserves until it is returned to light to resume photosynthesis. During the recovery period in the light, the cells are able to acclimate rapidly to current light levels and resume growth.

Size restoration by the auxospore that develops from the zygote is a crucial stage in diatom life cycles. However, information on sexual events in pelagic diatom species is very limited. We report for the first time auxospore formation by the pennate diatom Fragilariopsis kerguelensis (O'Hara) Hustedt during an iron-induced bloom in the Southern Ocean (EIFEX, European Iron Fertilization EXperiment). Auxospores of F. kerguelensis resembled those described for Pseudo-nitzschia species. The auxospore was characterized by an outer coating, the perizonium; two caps, one at each distal end; and four chloroplasts, one at each end and two in the central part. Different stages of auxospore elongation were recorded, with a length of 24–91 μm, but only the largest auxospores contained the initial cell, whose apical axis ranged between 76 and 90 μm. Gametangial cell walls were often attached to the auxospores and ranged from 10 to 31 μm in length. Auxospore abundances were consistently higher in the fertilized patch, where an increase in the F. kerguelensis population was observed, as compared with surrounding waters.

In Lake Suigetsu, central Japan, greenish/light-brown granules identified as cytoplasmic masses had been preserved in siliceous cell walls of freshwater diatoms in annual layers of lacustrine muds since the early Holocene. The lacustrine muds consisted of alternating dark-colored (rich in diatom valves, clay, and organic matter) and light-colored (mainly diatom valves) laminae. The greenish/light-brown granules were predominately preserved in frustules of the genus Aulacoseira preserved in the dark-colored laminae. The dark-colored laminae were inferred to have formed annually under stratified water caused by surface water warming in summer that caused the formation of an organic-rich anoxic layer on the lake bottom that favored granule preservation. The good preservation of cytoplasmic masses in dark-colored laminae suggested a cause for diatom assemblage periodicity, a phenomenon that was commonly noted in temperate lakes: the cells containing these masses could be potential seed stocks for subsequent spring blooms. Frustules of the most abundant granule-containing species, Aulacoseira nipponica (Skvortzow) Tuji, in the dark-colored laminae of the Early Holocene muds were abundant in the overlying light-colored laminae, suggesting that these species reproduced abundantly in springtime yielding a massive diatom bloom.

The bloom-forming cyanobacterium Microcystis aeruginosa Kutz 854 was cultured with 1.05 W·m−2 UV-B for 3 h every day, and its growth, pigments, and photosynthesis were investigated. The specific growth rates represented by chl a concentration and OD750 were inhibited 8% and 9% by UV-B exposure, respectively. Six days of UV-B treatment significantly reduced cellular contents of phycocyanin and allophycocyanin by 32% and 62%, respectively, and markedly increased the carotenoid content by 27%, but had little effect on the chl a content. The initial values of optimal photosynthetic efficiency for UV-B treated samples were, respectively, 52%, 87%, and 93% of controls on days 4, 7, and 10 of growth. The light-saturated photosynthetic rates at day 6 were significantly lower than controls grown without UV-B. The probability of electron transfer beyond QA decreased during UV-B exposure, and this indicated that the acceptor side of PSII was one of main damage sites. The adaptation of M. aeruginosa 854 to UV-B radiation could be observed from light-saturated photosynthetic rates on day 13 and diurnal changes of chl fluorescence during the late growth phase. When both exposed to higher UV-B, samples cultured under 1.05 W·m−2 UV-B for 9 days recovered faster than controls. It is suggested that M. aeruginosa 854 had at least three adaptive strategies to cope with the enhanced UV-B: increasing the synthesis of carotenoids to counteract reactive oxidants caused by UV-B exposure, degrading phycocyanin and allophycocyanin to avoid further damage to DNA and reaction centers, and enhancing the repair of UV-B induced damage to the photosynthetic apparatus.

In the St. Lawrence Estuary, annual recurrent blooms of the toxic dinoflagellate Alexandrium tamarense L. Balech are associated with brackish waters. Riverine inputs are suspected to favor bloom development by increasing water column stability and/or by providing growth stimulants such as humic substances (HS). A 17-day culture experiment was conducted to evaluate the importance of HS as growth factors for A. tamarense. Nonaxenic cultures were exposed to four HS extracts from three different sources: humic and fulvic acids isolated from the Manicouagan River, Quebec, Canada; humic acids from the Suwannee River, Georgia, United States; and a desalted alkaline soil extract. For each extract, four concentrations were tested as supplements to the artificial Keller medium, a nitrate-rich algal culture medium. Additions of HS from all sources significantly enhanced the overall growth rates relative to the controls. Concentrations of HS, estimated by UV spectrophotometry, remained constant throughout the exponential growth phase, suggesting that the HS were acting mainly as growth promoters during our experiment. Dose-response curves indicated that HS could increase the growth rate of A. tamarense even at low concentrations, such as those encountered in the St. Lawrence Estuary. Our results support the hypothesis that HS from the Manicouagan River plume can stimulate the development of toxic dinoflagellate blooms.

Molecular analyses and subsequent morphological reinvestigation of clonal isolates germinated from cysts previously assigned to Scrippsiella hangoei (Schiller) Larsen revealed considerable differences to vegetative cell isolates of this cold-water dinoflagellate from the northern Baltic Sea. The presence of hexagonal platelets on the cell surface and a characteristic acrobase on the episome agree with the description of Gymnodinium halophilum Biecheler. However, the arrangement of amphiesmal vesicles in more than nine latitudinal series indicates allocation of this dinoflagellate to Woloszynskia Thompson. We therefore reassign G. halophilum to Woloszynskia halophila. This species exhibits ultrastructural characteristics similar to Polarella glacialis Montresor et al. and symbiontic Gymnodinium Stein, such as stalked pyrenoids and a central eyespot consisting of multiple layers of crystal-filled vacuoles. A close relationship between these dinoflagellates is also supported by 28s rRNA sequence data. The preference for high salinities identifies W. halophila as a marine species. The spiny resting cysts of W. halophila are identical to the cysts formed during the massive encystment events previously attributed to S. hangoei in the Baltic Sea. This suggests that W. halophila is a significant contributor to the dinoflagellate spring blooms in the Baltic Sea. Scrippsiella hangoei clones, in turn, produce noncalcareous and smooth-walled cysts when crossed with a complementary mating type.

The dinoflagellate Alexandrium tamarense (Lebour) Balech 1985 is responsible for recurrent outbreaks of paralytic shellfish poisoning in the St. Lawrence Estuary. In July 1998, an A. tamarense red tide developed in the estuary with maximum cell concentrations reaching 2.3 × 10 6 cells.L - 1 in brackish surface waters. To estimate the growth rate of these cells, surface water samples from different locations and days during the bloom were incubated for 5 to 9 days under in situ temperature and light conditions. Growth rates varied both spatially and temporally between 0 and 0.55 day - 1 , reaching the maximum growth rate reported for this species in culture. High growth rates were measured even during the peak of the red tide, suggesting that the extremely high cell concentrations observed did not solely result from aggregation or physical concentration but also involved active cellular growth. Alexandrium tamarense cells were found over a large range of salinity (20.8-29.5 psu), but high densities and significant growth were only measured when salinity was lower than 24.5 psu. Under these conditions, the number of divisions achieved by A. tamarense was proportional to the amount of nitrate available at the beginning of the incubations, whereas variations in growth rate were apparently controlled by the availability of phosphate. We hypothesize that the ability of A. tamarense to perform vertical migrations and acquire nitrate at night pushes this species toward phosphate limitation in the St. Lawrence Estuary.

Benthic dinoflagellates of the genus Ostreopsis Schmidt are common in tropical and subtropical water, and some species produce toxins potentially involved in human intoxication events. A benthic bloom of Ostreopsis mascarenensis Quod was observed near Rodrigues Island during a survey of benthic dinoflagellates in the southwestern Indian Ocean. The morphology of O. mascarenensis was studied by LM and SEM. Preliminary screening of a crude extract of an O. mascarenensis bloom revealed neurotoxicity in mice similar to that induced by palytoxin. After partition of the crude extract, the highest toxicity was retained in the butanol-soluble fraction, which retained hemolytic activity suggestive of palytoxin analogues. Two new toxins, mascarenotoxin-A and -B, were resolved from this fraction by HPLC coupled to a diode array detector. The closed mass spectrum profile and fragmentation pattern obtained by advanced nano-electrospray ionization quadrupole time-of-flight mass spectrometry between purified toxins and a reference palytoxin confirmed the mascarenotoxins as palytoxin analogues. These results were confirmed by tandem mass spectrometry with the identification of specific fragment ion m/z 327. An on-line liquid chromatography protocol coupled to tandem mass spectrometry was developed for detection of these palytoxin analogues. The present study describes the first purification, chemical, and toxicological characterization of new palytoxin analogues isolated from a benthic bloom of O. mascarenensis. These results suggest that O. mascarenensis, which is largely distributed in the southwestern Indian Ocean, could be a source of palytoxin poisoning in this tropical area.

In autumn 2002, parasitic dinoflagellates of the genus Amoebophrya Koeppen infected populations of the bloom-forming thecate dinoflagellates Alexandrium affine (Inoue and Fukuyo) Balech and Gonyaulax spinifera (Claparede and Lachmann) Diesing in coastal waters of Korea. Here we present the first documentation of Amoebophrya infections in A. affine and G. spinifera and use host–parasite cultures to provide information on parasite development and total generation times. Parasites of the two dinoflagellate hosts differed in their site of infection, developing in the cytoplasm of A. affine but in the nucleus of G. spinifera. Developmental stages of the parasite strains were similar to the previous descriptions of Amoebophrya spp. infecting other dinoflagellates. A prominent feature of Amoebophrya infection in A. affine from natural field samples was the presence of abnormal “giant cells” in the long chains formed by this host species. The characteristic green autofluorescence of Amoebophrya infections was not evident under blue light excitation until very late in the infection cycle of the A. affine–Amoebophrya sp. system but was detectable throughout the infection cycle in the G. spinifera–Amoebophrya sp. system. Despite the relatively long duration (2–10 min) of the emergence process of Amoebophrya spp. from these thecate hosts, total parasite generation times were shorter (53–55 h) than those previously reported for athecate host–parasite systems. These observations provide the basis for better assessing the impact of parasitism by Amoebophrya sp. as a significant loss factor for thecate dinoflagellate populations in Korean waters.

In some lakes, large amounts of the potentially toxic cyanobacterium Microcystis overwinter in the sediment. This overwintering population might inoculate the water column in spring and promote the development of dense surface blooms of Microcystis during summer. In the Dutch Lake Volkerak, we found photochemically active Microcystis colonies in the sediment throughout the year. The most vital colonies originated from shallow sediments within the euphotic zone. We investigated whether recruitment of Microcystis colonies from the sediment to the water column was an active process, through production of gas vesicles or respiration of carbohydrate ballast. We calculated net buoyancy, as an indication of relative density, using the amounts and densities of the major cell constituents (carbohydrates, proteins, and gas vesicles). Carbohydrate content of benthic Microcystis cells was very low throughout the year. Buoyancy changes of benthic Microcystis were mostly a result of changes in gas vesicle volume. Before the summer bloom, net buoyancy and the amount of buoyant colonies in the sediment did not change. Therefore, recruitment of Microcystis from the sediment does not seem to be an active process regulated by internal buoyancy changes. Instead, our observations indicate that attachment of sediment particles to colonies plays an important part in the buoyancy state of benthic colonies. Therefore, we suggest that recruitment of Microcystis is more likely a passive process resulting from resuspension by wind-induced mixing or bioturbation. Consequently, shallow areas of the lake probably play a more important role in recruitment of benthic Microcystis than deep areas.

Alexandrium catenella (Whedon et Kofoid) Balech was isolated from Thau lagoon (northern Mediterranean) and its growth and uptake characteristics measured for nitrate, ammonium, and urea. Although affinity constants did not indicate a preference for ammonium over nitrate, there was a strong inhibition of nitrate uptake by ammonium when both nitrogen (N) sources were present. Nitrogen budgets during growth in cultures revealed major imbalances between decreases in dissolved N and increases in particulate N, indicating excretion of dissolved organic N during the early part of the growth phase and uptake during the later part. A quasi-unialgal bloom in November 2001 (4 × 10 6 cells . L - 1 ) allowed measurements of uptake of nitrate, nitrite, ammonium, and urea; net and gross growth rate of A. catenella; and grazing rates on this organism. The affinity constants indicate that it is not a strong competitor for the N nutrients tested when these are in low concentrations (

The smallest known eukaryote, Ostreococcus tauri Courties et Chretiennot-Dinet, was first reported as the dominant picoplankter in a French lagoon known for its diverse phytoplankton community and high oyster productivity. Long-term seasonal blooms of this picoeukaryote were observed in association with stable plankton communities. On 5 June 2001, a distinctive monotypic picoplankton bloom was detected by flow cytometry as part of an ongoing study of "brown tide" (Aureococcus anophagefferens) bloom initiation in Long Island bays. The bloom reached a concentration of 5 X 10 5 cells.mL - 1 in West Neck Bay and lasted less than 2 weeks. Epifluorescence microscopy and TEM indicated that the bloom organism was an Ostreococcus-like picoalga, the first ever observed in a Long Island bay. Many cells of this alga contained numerous virus-like particles. The Ostreococcus-like picoalga, which resembles O. tauri, was rare in samples collected the following week. Instead, a substantial increase in the Synechococcus population was observed. Such rapid population changes have not previously been reported for Ostreococcus. Viral lysis and grazing by heterotrophic nanoflagellates may have contributed to the rapid decline of the Ostreococcus-like cells in West Neck Bay.

Many shallow lakes in north temperate zones experience reduced dissolved oxygen concentration under ice. However, some shallow lakes display supersaturated dissolved oxygen concentrations (>20 mg.L - 1 ) in late winter under conditions of maximum ice thickness. During the winters of 1996, 1997, and 1999, we collected phytoplankton samples from Arrowwood Lake near Pingree, North Dakota to determine whether a specific alga was involved in dissolved oxygen supersaturation in this lake. Although dissolved oxygen supersaturation was not observed during this period, we did observe an increase in dissolved oxygen concentration that was associated with a phytoplankton bloom during late February and early March in both 1996 and 1997. In 1996, the bloom was composed of the dinoflagellate, Peridinium aciculiferum (Lemm.) Lemm. and several species of cryptomonads. A similar bloom of P. aciculiferum was followed by a bloom of several species of euglenoids in 1997. In contrast, P. aciculiferum was only a minor component of the winter phytoplankton, dissolved oxygen concentrations remained low, and no bloom event was observed in 1999. Statistical analyses indicated a significant relationship (r s = 0.57, P = 0.019) between dissolved oxygen levels and the density of the dinoflagellate, P. aciculiferum, but no significant relationship between dissolved oxygen levels and densities of other phytoplankton. These results suggest that the elevated levels of dissolved oxygen are associated with the dinoflagellate, P. aciculiferum. This bloom was most likely the result of an excystment event rather than a general growth response.

Microcystis aeruginosa Kutz. 7820 was cultured at 350 and 700 muL.L-1 CO2 to assess the impacts of doubled atmospheric CO2 concentration on this bloom-forming cyanobacterium. Doubling Of CO2 concentration in the airflow enhanced its growth by 52%-77%, with pH values decreased and dissolved inorganic carbon (DIC) increased in the medium. Photosynthetic efficiencies and dark respiratory rates expressed per unit chl a tended to increase with the doubling of CO2. However, saturating irradiances for photosynthesis and light-saturated photosynthetic rates normalized to cell number tended to decrease with the increase of DIC in the medium. Doubling of CO2 concentration in the airflow had less effect on DIC-saturated photosynthetic rates and apparent photosynthetic affinities for DIC. In the exponential phase, CO2 and HCO3- levels in the medium were higher than those required to saturate photosynthesis. Cultures with surface aeration were DIC limited in the stationary phase. The rate of CO2 dissolution into the liquid increased proportionally when CO2 in air was raised from 350 to 700 muL.L-1, thus increasing the availability of DIC in the medium and enhancing the rate of photosynthesis. Doubled CO2 could enhance CO2 dissolution, lower pH values, and influence the ionization fractions of various DIC species even when the photosynthesis was not DIC limited. Consequently, HCO3- concentrations in cultures were significantly higher than in controls, and the photosynthetic energy cost for the operation of CO2 concentrating mechanism might decrease.

Plankton samples from three inland embayments and several outer coastal sites of Washington State were collected from 1997 through 1999 and were examined for the presence of diatoms of the genus Pseudo-nitzschia and levels of the toxin, domoic acid (DA). Seven species were observed, including Pseudo-nitzschia pungens (Grunow ex Cleve) Hasle, P. multiseries (Hasle) Hasle, P. australis Frenguelli, P. fraudulenta (Cleve) Hasle, P. cf. heimii Manguim, P. pseudodelicatissima (Hasle) Hasle, and P. delicatissima (Cleve) Heiden. The coastal Pseudo-nitzschia species assemblages differed significantly from those observed within embayments. The dominant species observed at coastal sites were P. pseudodelicatissima and P. cf. heimii. Pseudo-nitzschia assemblages found in embayments included one or more of the following species: P. pungens, P. multiseries, P. australis, P. pseudodelicatissima, and P. fraudulenta. The nuclear large subunit rRNA gene was sequenced for six of the seven species identified. This sequence revealed that P. multiseries, P. pungens, P. australis, and P. heimii were genetically similar to those found in California, whereas P. delicatissima and P. pseudodelicatissima were distinct from the California isolates. Although the concentrations of DA in razor clams along Washington State coasts have exceeded regulatory limits several times since 1991, levels of DA in shellfish from Washington State embayments have not yet exceeded regulatory limits. The widespread presence of toxin-producing Pseudo-nitzschia species suggests, however, that toxic blooms are likely to occur within embayments in the future. In conjunction with the monitoring of environmental conditions conducive to toxic bloom formation, the development of species-specific probes for rapid and accurate detection of potentially toxic Pseudo-nitzschia species in this region would enable the forecasting of a toxic event before DA accumulates in shellfish, thereby reducing the impacts to coastal communities.

During a survey from January to March 1998, the occurrence of W-B radiation (UVBR)-induced DNA damage in Antarctic marine phytoplankton and bacterioplankton was investigated, Sampling was done in Ryder Bay, off the British base Rothera Station, 67 degreesS, 68 degreesW (British Antarctic Survey). Samples were taken regularly during the survey period at fixed depths, after which DNA damage was measured in various plankton size fractions: (> 10, 2-10, and 0,2-2 mum). Incident solar radiation was measured using spectroradiometry, whereas attenuation of biologically effective UVBR was studied using a DNA dosimeter, A diatom bloom was found in the bay during the research period, judging from microscopic observations and HPLC analyses of taxon-specific pigments, The high phytoplankton biomass likely caused strong attenuation of DNA effective UVBR (Kbd-eff). Kbd-eff values ranged from 0.83 .m(-1) at the peak of the bloom to 0.47 .m(-1) at the end of the season. UVBR-mediated DNA damage, as measured by cyclobutane pyrimidine dimer (CPD) abundance, was detected in all plankton Size fractions, Highest levels were found in the smallest size fraction, mainly consisting of heterotrophic bacteria. Clear CPD depth profiles were found during mid-summer (January, beginning of February) with surface levels exceeding 100 CPDs per million nucleotides in the bacterioplankton fraction. At that time, melting of the continuously present shelf ice caused strong salinity gradients in the upper meters, thereby stimulating water column stabilization. At the end of February and beginning of March, this phenomenon was less pronounced or absent, At that time, DNA damage was homogeneously distributed over the first 10 m, ranging between 20 and 30 CPDs per million nucleotides for the smallest size fraction

The haptophyte Phaeocystis antarctica Karsten is a dominant species within the seasonal bloom in the Ross Sea. One of the unique characteristics of this form is that carbon is partitioned between the cells and the colonial matrix, a relationship that is poorly documented for this region. We combined particulate organic carbon measurements and microscopic analysis of P. antarctica-dominated samples to assess the contribution of single cells, colony-associated cells, and mucilage to the carbon concentrations of waters with P. antarctica. Two cruises to the Ross Sea were completed, one in austral spring 1994 and one in summer 1995–1996. In 1994 the bloom was dominated by colonial P. antarctica that contributed up to 96% of the total autotrophic carbon, whereas in 1995–1996 a mixture of P. antarctica and diatoms occurred. P. antarctica colony volume (V ) was related to colonial cell number (NC) by the relationship V = 417 ×NC1.67. Total colony carbon (CCOL) was calculated as the sum of cell carbon (CCC) and mucus-related carbon (CM). We found the contribution of mucus carbon to be 213 ng C mm−3 of colony volume. For P. antarctica-dominated assemblages sampled at the peak of the bloom, CM represented a minor fraction (14 ± 4%) of colony carbon, and during early summer conditions CM was at most 33% of CCOL. This organism plays a cardinal role in the carbon cycle of many regions. These results constrain the partitioning of carbon between cellular material and the colony matrix, information that is necessary to accurately describe the biogeochemical cycles influenced by this species.

Laboratory experiments with batch cultures showed that the Texas brown tide alga, Aureoumbra lagunensis Stockwell, DeYoe, Hargraves et Johnson, produced a large amount of extracellular polymeric substance (EPS) that formed an extracellular polysaccharide mucus layer. Both dissolved and particulate carbohydrate and EPS concentrations increased as A. lagunensis cultures progressed from exponential growth phase, through stationary phase, to declining phase. Particulate carbohydrate and EPS concentrations per cell were more than three times higher during the declining phase than that in exponential growth phase, reflecting a large increase in the EPS mucus layer. The amounts of carbohydrate and EPS produced by A. lagunensis were significantly higher under hypersaline conditions. The thicker EPS mucus layer surrounding A. lagunensis cells under hypersaline conditions might be a protective adaptation that permits it to bloom under hypersaline conditions that most other phytoplankton cannot survive. This could be one of the reasons why the Texas brown tide persisted in the Laguna Madre, an often hypersaline coastal lagoon, for 7 years.

A toxic phytoplankton bloom, dominated by the prymnesiophyte Chrysochromulina leadbeateri Estep, developed in the Ofotfjord–Tysfjord area (North Norway) in mid-May and ended in late June 1991 in Vestfjorden and the adjacent fjord areas. Chrysochromulina leadbeateri dominated at total cell densities of >2 × 106 cells·L−1; at lower total cell densities, C. leadbeateri was accompanied by other Chrysochromulina species, peridinin-containing dinoflagellates, and diatoms. Bio-optical characteristics and pigmentation in laboratory and field strains of C. leadbeateri allowed for the interpretation of the optical signatures within the bloom. The bio-optical data suggested healthy and actively growing cells during the bloom. About 600 metric tons of pen-raised Atlantic salmon were killed by the C. leadbeateri bloom. A laboratory study was conducted to assess the potential impact of finfish on C. leadbeateri growth. It was found that the polyamine putrescine enhanced cell biomass and hemolytic activity. Given this, a possible scenario for the development of this bloom and the level of toxicity is hypothesized: (1) The nutrient loading in the Ofotfjord area was enhanced during the winter of 1990–1991 due to the overwintering of 1.5 × 106 metric tons of herring from a depth of 0–250 m. This may have sustained a large stock of the mixotrophic C. leadbeateri in early spring before light regime (irradiance, spectral irradiance, and day length) made net photosynthesis possible. (2) The release of polyamines during the decay of dead fish (e.g. putrescine, cadaverine, and histamine) may have acted as cofactors with ichthyotoxins making “hypertoxic complexes” with the polyamines enhancing growth in the mixotrophic C. leadbeateri.

In estuaries, phytoplankton are exposed to rapidly changing conditions that may have profound effects on community structure and function. In these experiments, we evaluated the growth, productivity, and compositional responses of natural phytoplankton communities exposed to limiting nutrient additions and incubation conditions typical of estuarine habitats. Mesocosm bioassays were used to measure the short-term (2-day) growth rate, primary productivity, and group-specific biomass responses of the phytoplankton community in the Neuse River Estuary, North Carolina. A three-factor (mixing, sediment addition, and nutrient addition) experimental design was applied using 55-L mesocosm tanks. Growth rates were determined using the 14C photopigment radiolabeling method, and the abundance of algal groups was based on quantification of chemosystematic photopigments by HPLC. For Neuse River Estuary phytoplankton communities, stratified (nonmixed), turbid, and low-nitrate conditions favored increases in cryptomonad biomass. Mixed, turbid, high-nitrate conditions were favorable for increased primary productivity and chlorophytes, diatoms, and cyanobacteria. The highest community growth rates occurred under calm, high-nitrate conditions. This approach provided an assessment of the community-level phytoplankton responses and insights into the mechanisms driving blooms and bloom species in estuarine waters. The ability to rapidly alter growth rates to capitalize on conditions conducive for growth may play an important role in the timing, extent, and species involved with blooms in estuarine waters. Adaptive growth rate responses of individual species, as well as the community as a whole, further illustrate the sensitivity of estuarine ecosystems to excessive N inputs.