Environmental control of harmful dinoflagellates and diatoms in a fjordic system

Fjordic coastlines provide an ideal protected environment for both finfish and shellfish aquaculture operations. This study reports the results of a cruise to the Scottish Clyde Sea, and associated fjordic sea lochs, that coincided with blooms of the diarrhetic shellfish toxin producing dinoflagellate Dinophysis acuta and the diatom genus Chaetoceros, that can generate finfish mortalities. Unusually, D. acuta reached one order of magnitude higher cell abundance in the water column (2840 cells L⁻¹) than the more common Dinophysis acuminata (200 cells L⁻¹) and was linked with elevated shellfish toxicity (maximum 601 ± 237 μg OA eq/kg shellfish flesh) which caused shellfish harvesting closures in the region. Significant correlations between D. acuta abundance and that of Mesodinium rubrum were also observed across the cruise transect potentially supporting bloom formation of the mixotrophic D. acuta. Significant spatial variability in phytoplankton that was related to physical characteristics of the water column was observed, with a temperature-driven frontal region at the mouth of Loch Fyne being important in the development of the D. acuta, but not the Chaetoceros bloom. The front also provided important protection to the aquaculture located within the loch, with neither of the blooms encroaching within it. Analysis based on a particle-tracking model confirms the importance of the front to cell transport and shows significant inter-annual differences in advection within the region, that are important to the harmful algal bloom risk therein.     

Toxicity studies of Trichodesmium erythraeum (Ehrenberg, 1830) bloom extracts,from Phoenix Bay,Port Blair,Andamans

Occurrence of toxic cyanobacterial blooms has become a worldwide problem, increasing the risk of human poisoning due to consumption of seafood contaminated with cyanotoxins. Though no such cases of human intoxication due to toxic blooms have been reported so far from India, most of the studies related to blooms have been restricted to reporting of a bloom and/or antimicrobial activity of its extract. Detailed toxicity study of cyanobacterial blooms are lacking. A study on the toxicity of a dense bloom (14.56 × 10⁶ trichomes L⁻¹) of the marine diazotrophic cyanobacteria, Trichodesmium erythraeum, observed in the coastal waters of Phoenix Bay, Port Blair, Andamans was undertaken. The significance of this bloom is that it was a single species and had conspicuously inhibited the growth of other phytoplankton and complete exclusion of zooplankton from the bloom region, intimating the involvement of toxins in the bloom. The cyanobacterial extracts showed prominent antimicrobial activity against certain human pathogenic bacteria and fungi. Studies on the toxicity of the cyanobacterial extracts was carried out using brine shrimp bioassay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and comet assay. The cyanobacterial extract exhibited toxic effect to Artemia salina causing mortality of up to 40% after 48 h at a concentration of 1 mg mL⁻¹, while it induced cytotoxicity in cell lines (HepG2 and HaCat) and caused DNA damage in human lymphocytes in vitro.     

Toward predicting Dinophysis blooms off NW Iberia: A decade of events

Dinophysis acuminata and Dinophysis acuta are recurrent species off NW Iberia but their outbreaks occur under different conditions. A decade (2004–2013) of weekly data for each species at two sentinel stations located at the entrance of Rias de Aveiro-AV (NW Portugal, 40°38.6′ N) and Pontevedra-PO (Galicia, Spain, 42°21.5′ N), were used to investigate the regional synchronism and mesoscale differences related to species detection, bloom (>200 cells L⁻¹) initiation and development. Results highlight the high interannual variability of bloom events and summarize the associated meteorological/oceanographic conditions. D. acuta blooms were observed in 2004–2008 and 2013, and the species highest maxima at AV occurred after the highest maxima of its prey Mesodinium, with a time-lag of 2–3 weeks. D. acuminata blooms were observed every year at both stations. The cell concentration time series shows that the blooms generally present a sequence starting in March with D. acuminata in PO and three weeks later in AV, followed by D. acuta that starts at AV and three months later in PO. Exceptionally, D. acuminata blooms occurred earlier at AV than PO, namely in high spring upwelling (2007) or river runoff (2010) years. A four-year gap (2009–2012) of D. acuta blooms occurred after an anomalous 2008 autumn with intense upwelling which is interpreted as the result of an equatorward displacement of the population core. Numerical model solutions are used to analyze monthly alongshore current anomalies and test transport hypotheses for selected events. The results show a strong interannual variability in the poleward/equatorward currents associated with changes in upwelling forcing winds, the advection of D. acuta blooms from AV to PO and the possibility that D. acuminata blooms at AV might result from inocula advected southward from PO. However, the sensitivity of the results to vertical position of the lagrangian tracers call for more studies on species distribution at the various bloom stages.     

The binding of okadaic acid analogs to recombinant OABP2.1 originally isolated from the marine sponge Halichondria okadai

The binding between [24-³H]okadaic acid (OA) and a recombinant OA binding protein OABP2.1 was examined using various OA analog, including methyl okadaate, norokadanone, 7-deoxy OA, and 14,15-dihydro OA, 7-O-palmitoyl DTX1, to investigate the structure activity relationship. Among them, 7-O-palmitoyl DTX1, which is one of the diarrhetic shellfish poisoning (DSP) toxins identified in shellfish, displayed an IC₅₀ for [24-³H]OA binding at 51 ± 6.3 nM (Mean ± SD). In addition, a synthetic compound, N-pyrenylmethyl okadamide, exhibited its IC₅₀ at 10 ± 2.9 nM (Mean ± SD). These results suggested that the recombinant OABP2.1 and the N-pyrenylmethyl okadamide might be core substances in a novel assay for the DSP toxins.     

Production of dinophysistoxin-1 and pectenotoxin-2 by a culture of Dinophysis acuminata (Dinophyceae)

The production of diarrhetic shellfish poisoning toxins (okadaic acid analogues and other lipophilic toxins) by a culture of Dinophysis acuminata, fed with the autotrophic ciliate Myrionecta rubra, was confirmed by LC–MS analysis, and the toxin profile compared with that in the field assemblage of the same species. The growth response of D. acuminata to the density of the food organism was also examined in laboratory experiments. In semi-continuous culture experiments, the growth rates of D. acuminata increased with increasing density of M. rubra and a maximum growth rate of 0.67 per day was calculated. In batch culture experiments; the cellular content of PTX2 and DTX1 were 14.7–14.8 and 2.5–4.8 pg cell^?1, respectively. Okadaic acid, dinophysistoxin-3, pectenotoxin-1, pectenotoxin-6, yessotoxin (YTX) and 45-OHYTX were not detected. PTX2 was detected (cellular toxin content: 22 pg cell^?1), but DTX1 was not detected, in an extract of D. acuminata collected from natural seawater at the same location where the cultured D. acuminata specimens were isolated. These results strongly suggest that D. acuminata produces these toxins during cell growth and that environmental factors influence variations in the toxin composition and specific cellular toxicity.     

Toxigenic phytoplankton and concomitant toxicity in the mussel Choromytilus meridionalis off the west coast of South Africa

The variability of toxigenic phytoplankton and the consequent uptake and loss of toxins by the mussel Choromytilus meridionalis was investigated in the southern Benguela at the event scale (3–10 days) in response to the upwelling–downwelling cycle. Phytoplankton and mussel samples were collected daily (20 March–11 April 2007) from a mooring station (32.04°S; 18.26°E) located 3.5 km offshore of Lambert's Bay, within the St Helena Bay region. Rapid changes in phytoplankton assemblages incorporated three groups of toxigenic phytoplankton: (1) the dinoflagellate Alexandrium catenella; (2) several species of Dinophysis, including Dinophysis acuminata, Dinophysis fortii, Dinophysis hastata and Dinophysis rotundata; and (3) members of the diatom genus Pseudo-nitzschia. Analysis of phytoplankton concentrates by LC–MS/MS or LC-FD provided information on the toxin composition and calculated toxicity of each group. Several additional in vitro assays were used for the analysis of toxins in mussels (ELISA, RBA, MBA for PSP toxins; and ELISA for DSP toxins). Good correspondence was observed between methods except for the MBA, which provided significantly lower (approximately 2-fold) estimates of PSP toxins. PSP and DSP toxins both exceeded the regulatory limits in Choromytilis meridionalis, but ASP toxins were undetected. Differences were observed in the composition of both PSP and DSP toxins in C. meridionalis from that of the ingested dinoflagellates (PSP toxins showed an increase in STX, C1,2, and traces of dcSTX and GTX1,4 and a decrease in NEO; DSP toxins showed an increased in DTX1, and traces of PTX2sa, and a decrease in OA). The rate of loss of PSP toxins following dispersal of the A. catenella boom was 0.12 d⁻¹. Variation in the loss rates of different PSP toxins contributed to the change in toxin profile in C. meridionalis. Prediction of net toxicity in shellfish of the nearshore environment in the southern Benguela is limited due to rapid phytoplankton community changes, high variability in cellular toxicity, and the selective uptake and loss of toxins, and/or transformation of toxins.     

Alteration of plankton communities and biogeochemical cycles by harmful Cochlodinium polykrikoides (Dinophyceae) blooms

Cochlodinium polykrikoides is a globally distributed, ichthyotoxic, bloom-forming dinoflagellate. Blooms of C. polykrikoides manifest themselves as large (many km²) and distinct patches with cell densities exceeding 10³ ml⁻¹ while water adjacent to these patches can have low cell densities (<100 cells ml⁻¹). While the effect of these blooms on fish and shellfish is well-known, their impacts on microbial communities and biogeochemical cycles are poorly understood. Here, we investigated plankton communities and the cycling of carbon, nitrogen, and B-vitamins within blooms of C. polykrikoides and compared them to areas in close proximity (<100 m) with low C. polykrikoides densities. Within blooms, C. polykrikoides represented more than 90% of microplankton (>20 μm) cells, and there were significantly more heterotrophic bacteria and picoeukaryotic phytoplankton but fewer Synechococcus. Terminal restriction fragment length polymorphism analysis of 16S and 18S rRNA genes revealed significant differences in community composition between bloom and non-bloom samples. Inside the bloom patches, concentrations of vitamin B₁₂ were significantly lower while concentrations of dissolved oxygen were significantly higher. Carbon fixation and nitrogen uptake rates were up to ten times higher within C. polykrikoides bloom patches. Ammonium was a more important source of nitrogen, relative to nitrate and urea, for microplankton within bloom patches compared to non-bloom communities. While uptake rates of vitamin B₁ were similar in bloom and non-bloom samples, vitamin B₁₂ was taken up at rates five-fold higher (>100 pmol⁻¹ L⁻¹ d⁻¹) in bloom samples, resulting in turn-over times of hours during blooms. This high vitamin demand likely led to the vitamin B₁₂ limitation of C. polykrikoides observed during nutrient amendment experiments conducted with bloom water. Collectively, this study revealed that C. polykrikoides blooms fundamentally change microbial communities and accelerate the cycling of carbon, some nutrients, and vitamin B₁₂.     

Detection of Karenia brevis blooms on the west Florida shelf using in situ backscattering and fluorescence data

Using shipboard data collected from the central west Florida shelf (WFS) between 2000 and 2001, an optical classification algorithm was developed to differentiate toxic Karenia brevis blooms (>10⁴ cells l⁻¹) from other waters (including non-blooms and blooms of other phytoplankton species). The identification of K. brevis blooms is based on two criteria: (1) chlorophyll a concentration ≥1.5 mg m⁻³ and (2) chlorophyll-specific particulate backscattering at 550 nm ≤ 0.0045 m² mg⁻¹. The classification criteria yielded an overall accuracy of 99% in identifying both K. brevis blooms and other waters from 194 cruise stations. The algorithm was validated using an independent dataset collected from both the central and south WFS between 2005 and 2006. After excluding data from estuarine and post-hurricane turbid waters, an overall accuracy of 94% was achieved with 86% of all K. brevis bloom data points identified successfully. Satisfactory algorithm performance (88% overall accuracy) was also achieved when using underway chlorophyll fluorescence and backscattering data collected during a repeated alongshore transect between Tampa Bay and Florida Bay in 2005 and 2006. These results suggest that it may be possible to use presently available, commercial optical backscattering instrumentation on autonomous platforms (e.g. moorings, gliders, and AUVs) for rapid and timely detection and monitoring of K. brevis blooms on the WFS.     

Climate variability and Dinophysis acuta blooms in an upwelling system

Dinophysis acuta is a frequent seasonal lipophilic toxin producer in European Atlantic coastal waters associated with thermal stratification. In the Galician Rías, populations of D. acuta with their epicentre located off Aveiro (northern Portugal), typically co-occur with and follow those of Dinophysis acuminata during the upwelling transition (early autumn) as a result of longshore transport. During hotter than average summers, D. acuta blooms also occur in August in the Rías, when they replace D. acuminata. Here we examined a 30-year (1985–2014) time series of D. acuta from samples collected by the same method in the Galician Rías. Our main objective was to identify patterns of distribution and their relation with climate variability, and to explain the exceptional summer blooms of D. acuta in 1989–1990. A dome-shaped relationship was found between summer upwelling intensity and D. acuta blooms; cell maxima were associated with conditions where the balance between upwelling intensity and heating, leading to deepened thermoclines, combined with tidal phase (3 days after neap tides) created windows of opportunity for this species. The application of a generalized additive model based on biological (D. acuta inoculum) and environmental predictors (Cumulative June–August upwelling CUI_JJA, average June–August SST_JJA and tidal range) explained more than 70% of the deviance for the exceptional summer blooms of D. acuta, through a combination of moderate (35,000–50,000 m³ s⁻¹ km⁻¹) summer upwelling (CUI_JJA), thermal stratification (SST_JJA > 17 °C) and moderate tidal range (∼2.5 m), provided D. acuta cells (inoculum) were present in July. There was no evidence of increasing trends in D. acuta bloom frequency/intensity nor a clear relationship with NAO or other long-term climatic cycles. Instead, the exceptional summer blooms of 1989–1990 appeared linked to extreme hydroclimatic anomalies (high positive anomalies in SST and NAO index), which affected most of the European Atlantic coast.     

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 ± 0.03 pg/cell, 0.44 ± 0.05 pg/cell), DTX1 (0.45 ± 0.09 pg/cell, 0.64 ± 0.10 pg/cell) and PTX2 (10.4 ± 1.4 pg/cell, 11.0 ± 1.9 pg/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.     

Potential impact of an exceptional bloom of Karenia mikimotoi on dissolved oxygen levels in waters off western Ireland

In the summer of 2005 an exceptional bloom of the dinoflagellate Karenia mikimotoi occurred along Ireland's Atlantic seaboard and was associated with the mass mortality of both benthic and pelagic marine life. Oxygen depletion, cellular toxicity and physical smothering, are considered to be the main factors involved in mortality. In this paper we use a theoretical approach based on stoichiometry (the Anderson ratio) and an average K. mikimotoi cellular carbon content of 329 pg C cell⁻¹ (n = 20) to calculate the carbonaceous and nitrogenous oxygen demand following bloom collapse. The method was validated against measurements of biochemical oxygen demand and K. mikimotoi cell concentration. The estimated potential oxygen utilisation (POU) was in good agreement with field observations across a range of cell concentrations. The magnitude of POU following bloom collapse, with the exception of three coastal areas, was considered insufficient to cause harm to most marine organisms. This indicates that the widespread occurrence of mortality was primarily due to other factors such as cellular toxicity and/or mucilage production, and not oxygen depletion or related phenomena. In Donegal Bay, Kilkieran Bay and inner Dingle Bay, where cell densities were in the order of 10⁶ cells L⁻¹, estimated POU was sufficient to cause hypoxia. Of the three areas, Donegal Bay is considered to be the most vulnerable due to its hydrographic characteristics (seasonally stratified, weak residual flow) and hypoxic conditions (2.2 mg L⁻¹ O₂) were directly observed in the Bay post bloom collapse. Here, depending on the time of bloom collapse, depressed DO levels could persist for weeks and continue to have a potentially chronic impact on the Bay.     

Seasonal variability of lipophilic toxins during a Dinophysis acuta bloom in Western Iberia: Differences between picked cells and plankton concentrates

This work describes and compares the seasonal variability of toxin profiles and content, estimated by LC–MS analyses, in picked cell of Dinophysis acuta Ehrenberg, in plankton concentrates rich in this species, and in extracellular lipophilic toxins collected by adsorbent resins during weekly sampling in a Galician ría (Western Iberia) from October 2005 to January 2006. Picked cells of D. acuta—which exhibited a fairly stable OA:DTX2 ratio, close to 3:2, but a variable okadaates:PTX2 ratio—showed a 9-fold variation in cell toxin quota, which was partly related to cellular volume, with maximum values (19 pg cell⁻¹) observed during the exponential decline of the population. Large differences in toxin profiles and content were observed between picked cells and plankton concentrates (up to 73 pg cell⁻¹ in the latter), that were most conspicuous after the bloom decline. The toxin profile of picked cells was more similar to that observed in the adsorbent resins than to the profiles of plankton concentrates. Their continued detection several weeks after the disappearance of Dinophysis spp. indicates that these toxins may take a long time to be degraded. It is concluded that analyses of picked-cells are essential to determine the contribution of each species of Dinophysis to a toxic outbreak. Estimates of cellular toxin content from plankton concentrates can lead to considerable overestimates after Dinophysis blooms decay due to extracellular toxins that persist in the water column, possibly bound to organic aggregates and detritus, and are retained (>0.22 μm) in the filters.     

Characterization of multiple isolates from an Alexandrium ostenfeldii bloom in The Netherlands

Alexandrium ostenfeldii is an emerging harmful algal bloom species forming a global threat to coastal marine ecosystems, with consequences for fisheries and shellfish production. The Oosterschelde estuary is a shallow, macrotidal and mesotrophic estuary in the southwest of The Netherlands with large stocks of mussels, oysters, and cockles. These shellfish stocks were threatened by a recent A. ostenfeldii bloom in the Ouwerkerkse Kreek, which is a brackish water creek discharging water into the Oosterschelde. Little is yet known about the characteristics of the A. ostenfeldii population in this creek. We therefore isolated 20 clones during an A. ostenfeldii bloom in 2013, and characterized these clones on their growth and toxin profile in their exponential growth phase. The cyclic imines were identified by comparison of A. ostenfeldii extracts with the retention time and CID spectra of standard solutions, or with published CID spectra. We furthermore assessed the allelochemical potency and phylogeny of a selection of 10–12 clones. Morphology and molecular phylogeny showed that all clones belong to Group 1 of A. ostenfeldii. All clones showed comparable growth rates of on average 0.22 ± 0.03 d⁻¹. During exponential growth, they all produced a unique combination of paralytic shellfish poisoning toxins, spirolides and gymnodimines, of which particularly the latter showed a high intra-specific variability, with a 25-fold difference between clones with the lowest and highest cell quota. Furthermore, the selected 12 clones showed high allelopathic potencies with EC₅₀ values based on lysis assays against the cryptophyte Rhodomonas salina between 212 and 525 A. ostenfeldii cells mL⁻¹. Lytic activities were lower for cell extracts, indicating an important extracellular role of these compounds. A high intra-specific variability may add to the success of genotypically diverse A. ostenfeldii blooms, and make populations resilient to changes in environmental and climatic conditions.     

Effects of hydrology and river management on the distribution,abundance and persistence of cyanobacterial blooms in the Murray River,Australia

Major cyanobacterial blooms (biovolume > 4 mm³ L⁻¹) occurred in the main water reservoirs on the upper Murray River, Australia during February and March 2010. Cyanobacterial-infested water was released and contaminated rivers downstream. River flow velocities were sufficiently high that in-stream bloom development was unlikely. The location has a temperate climate but experienced drought in 2010, causing river flows that were well below the long-term median values. This coupled with very low bed gradients meant turbulence was insufficient to destroy the cyanobacteria in-stream. Blooms in the upper 500 km of the Murray and Edward Rivers persisted for 5 weeks, but in the mid and lower Murray blooms were confined to a small package of water that moved progressively downstream for another 650 km. Anabaena circinalis was the dominant species present, confirmed by 16S rRNA gene sequencing, but other potentially toxic species were also present in smaller amounts. Saxitoxin (sxtA), microcystin (mcyE) and cylindrospermopsin (aoaA) biosynthesis genes were also detected, although water sample analysis rarely detected these toxins. River water temperature and nutrient concentrations were optimal for bloom survival. The operational design of weirs and retention times within weir pools, as well as tributary inflows to and diversions from the Murray River all influenced the distribution and persistence of the blooms. Similar flow, water quality and river regulation factors were underlying causes of another bloom in these rivers in 2009. Global climate change is likely to promote future blooms in this and other lowland rivers.     

Monitoring,management, and mitigation of Karenia blooms in the eastern Gulf of Mexico

Annual blooms of the toxic dinoflagellate Karenia brevis in the eastern Gulf of Mexico represent one of the most predictable global harmful algal bloom (HAB) events, yet remain amongst the most difficult HABs to effectively monitor for human and environmental health. Monitoring of Karenia blooms is necessary for a variety of precautionary, management and predictive purposes. These include the protection of public health from exposure to aerosolized brevetoxins and the consumption of toxic shellfish, the protection and management of environmental resources, the prevention of bloom associated economic losses, and the evaluation of long term ecosystem trends and for potential future bloom forecasting and prediction purposes. The multipurpose nature of Karenia monitoring, the large areas over which blooms occur, the large range of Karenia cell concentrations (from 5 × 10³ cells L^?1 to >1 × 10⁶ cells L^?1) over which multiple bloom impacts are possible, and limitations in resources and knowledge of bloom ecology have complicated K. brevis monitoring, mitigation and management strategies. Historically, K. brevis blooms were informally and intermittently monitored on an event response basis in Florida, usually in the later bloom stages after impacts (e.g. fish kills, marine mammal mortalities, respiratory irritation) were noted and when resources were available. Monitoring of different K. brevis bloom stages remains the most practical method for predicting human health impacts and is currently accomplished by the state of Florida via direct microscopic counts of water samples from a state coordinated volunteer HAB monitoring program. K. brevis cell concentrations are mapped weekly and disseminated to stakeholders via e-mail, web and toll-free phone numbers and provided to Florida Department of Agriculture and Consumer Services (FDACS) for management of both recreational and commercial shellfish beds in Florida and to the National Oceanic and Atmospheric Administration (NOAA) for validation of the NOAA Gulf of Mexico HAB bulletin for provision to environmental managers. Many challenges remain for effective monitoring and management of Karenia blooms, however, including incorporating impact specific monitoring for the diverse array of potential human and environmental impacts associated with blooms, timely detection of offshore bloom initiation, sampling of the large geographic extent of blooms which often covers multiple state boundaries, and the involvement of multiple Karenia species other than K. brevis (several of which have yet to be isolated and described) with unknown toxin profiles. The implementation and integration of a diverse array of optical, molecular and hybrid Karenia detection technologies currently under development into appropriate regulatory and non-regulatory monitoring formats represents a further unique challenge.     

Controls on the initiation and development of blooms of the dinoflagellate Cochlodinium polykrikoides Margalef in lower Chesapeake Bay and its tributaries

Massive blooms of the dinoflagellate Cochlodinium polykrikoides occur annually in the Chesapeake Bay and its tributaries. The initiation of blooms and their physical transport has been documented and the location of bloom initiation was identified during the 2007 and 2008 blooms. In the present study we combined daily sampling of nutrient concentrations and phytoplankton abundance at a fixed station to determine physical and chemical controls on bloom formation and enhanced underway water quality monitoring (DATAFLOW) during periods when blooms are known to occur. While C. polykrikoides did not reach bloom concentrations until late June during 2009, vegetative cells were present at low concentrations in the Elizabeth River (4 cells ml⁻¹) as early as May 27. Subsequent samples collected from the Lafayette River documented the increase in C. polykrikoides abundance in the upper branches of the Lafayette River from mid-June to early July, when discolored waters were first observed. The 2009 C. polykrikoides bloom began in the Lafayette River when water temperatures were consistently above 25 °C and during a period of calm winds, neap tides, high positive tidal residuals, low nutrient concentrations, and a low dissolved inorganic nitrogen (DIN) to dissolved inorganic phosphorous (DIP) ratio. The pulsing of nutrients associated with intense but highly localized storm activity during the summer months when water temperatures are above 25 °C may play a role in the initiation of C. polykrikoides blooms. The upper Lafayette River appears to be an important area for initiation of algal blooms that then spread to other connected waterways.     

Nutrient ratios influence variability in Pseudo-nitzschia species diversity and particulate domoic acid production in the Bay of Seine (France)

The population dynamics of different Pseudo-nitzschia species, along with particulate domoic acid (pDA) concentrations, were studied from May 2012 to December 2013 in the Bay of Seine (English Channel, Normandy). While Pseudo-nitzschia spp. blooms occurred during the two years of study, Pseudo-nitzschia species diversity and particulate domoic acid concentrations varied greatly. In 2012, three different species were identified during the spring bloom (P. australis, P. pungens and P. fraudulenta) with high pDA concentrations (∼1400 ng l⁻¹) resulting in shellfish harvesting closures. In contrast, the 2013 spring was characterised by a P. delicatissima bloom without any toxic event. Above all, the results show that high pDA concentrations coincided with the presence of P. australis and with potential silicate limitation (Si:N < 1), while nitrate concentrations were still replete. The contrasting environmental conditions between 2012 and 2013 highlight different environmental controls that might favour the development of either P. delicatissima or P. australis. This study points to the key role of Pseudo-nitzschia diversity and cellular toxicity in the control of particulate domoic acid variations and highlights the fact that diversity and toxicity are influenced by nutrients, especially nutrient ratios.     

Recurrent vernal presence of the toxic Alexandrium tamarense/Alexandrium fundyense (Dinoflagellata) species complex in Narragansett Bay,USA

The vernal occurrence of toxic dinoflagellates in the Alexandrium tamarense/Alexandrium fundyense species complex in an enclosed embayment of Narragansett Bay (Wickford Cove, Rhode Island) was documented during 2005 and 2009–2012. This is the first report of regular appearance of the Alexandrium fundyense/Alexandrium tamarense species complex in Narragansett Bay. Thecal plate analysis of clonal isolates using SEM revealed cells morphologically consistent with both Alexandrium tamarense Lebour (Balech) and Alexandrium fundyense Balech. Additionally, molecular analyses confirmed that the partial sequences for 18S through the D1–D2 region of 28S were consistent with the identity of the two Alexandrium species. Toxin analyses revealed the presence of a suite of toxins (C1/2, B1 (GTX-5), STX, GTX-2/3. Neo, and GTX-1/4) in both Alexandrium tamarense (6.31 fmol cell⁻¹ STX equiv.) and Alexandrium fundyense (9.56 fmol cell⁻¹ STX equiv.) isolated from Wickford Cove; the toxicity of a Narragansett Bay Alexandrium peruvianum isolate (1.79 fmol cell⁻¹ STX equiv.) was also determined. Combined Alexandrium tamarense/Alexandrium fundyense abundance in Wickford Cove reached a peak abundance of 1280 cells L⁻¹ (May of 2010), with the combined abundance routinely exceeding levels leading to shellfishing closures in other systems. The toxic Alexandrium tamarense/Alexandrium fundyense species complex appears to be a regular component of the lower Narragansett Bay phytoplankton community, either newly emergent or previously overlooked by extant monitoring programs.     

Termination of a toxic Alexandrium bloom with hydrogen peroxide

The dinoflagellate Alexandrium ostenfeldii is a well-known harmful algal species that can potentially cause paralytic shellfish poisoning (PSP). Usually A. ostenfeldii occurs in low background concentrations only, but in August of 2012 an exceptionally dense bloom of more than 1 million cells L⁻¹ occurred in the brackish Ouwerkerkse Kreek in The Netherlands. The A. ostenfeldii bloom produced both saxitoxins and spirolides, and is held responsible for the death of a dog with a high saxitoxin stomach content. The Ouwerkerkse Kreek routinely discharges its water into the adjacent Oosterschelde estuary, and an immediate reduction of the bloom was required to avoid contamination of extensive shellfish grounds. Previously, treatment of infected waters with hydrogen peroxide (H₂O₂) successfully suppressed cyanobacterial blooms in lakes. Therefore, we adapted this treatment to eradicate the Alexandrium bloom using a three-step approach. First, we investigated the required H₂O₂ dosage in laboratory experiments with A. ostenfeldii. Second, we tested the method in a small, isolated canal adjacent to the Ouwerkerkse Kreek. Finally, we brought 50 mg L⁻¹ of H₂O₂ into the entire creek system with a special device, called a water harrow, for optimal dispersal of the added H₂O₂. Concentrations of both vegetative cells and pellicle cysts declined by 99.8% within 48 h, and PSP toxin concentrations in the water were reduced below local regulatory levels of 15 μg L⁻¹. Zooplankton were strongly affected by the H₂O₂ treatment, but impacts on macroinvertebrates and fish were minimal. A key advantage of this method is that the added H₂O₂ decays to water and oxygen within a few days, which enables rapid recovery of the system after the treatment. This is the first successful field application of H₂O₂ to suppress a marine harmful algal bloom, although Alexandrium spp. reoccurred at lower concentrations in the following year. The results show that H₂O₂ treatment provides an effective emergency management option to mitigate toxic Alexandrium blooms, especially when immediate action is required.