Three-dimensional structure of a Karenia brevis bloom: Observations from gliders,satellites, and field measurements

Autonomous underwater gliders with customized sensors were deployed in October 2011 on the central West Florida Shelf to measure a Karenia brevis bloom, which was captured in satellite imagery since late September 2011. Combined with in situ taxonomy data, satellite measurements, and numerical circulation models, the glider measurements provided information on the three-dimensional structure of the bloom. Temperature, salinity, fluorescence of colored dissolved organic matter (CDOM) and chlorophyll-a, particulate backscattering coefficient, and K. brevis-specific chlorophyll-a concentrations were measured by the gliders over >250 km from the surface to about 30-m water depth on the shallow shelf. At the time of sampling the bloom was characterized by uniform vertical structures, with relatively high chlorophyll-a and CDOM fluorescence, low temperature, and high salinity. Satellite data extracted along the glider tracks demonstrated coherent spatial variations as observed by the gliders. Further, the synoptic satellite observations revealed the bloom evolution during the 7 months between late September 2011 and mid April 2012, and showed the maximum bloom size of ∼3000 km² around 23 November. The combined satellite and in situ data also confirmed that the ratio of satellite-derived fluorescence line height (FLH) to particulate backscattering coefficient at 547 nm (b_bp(547)) could be used as a better index than FLH alone to detect K. brevis blooms. Numerical circulation models further suggested that the bloom could have been initiated offshore and advected onshore via the bottom Ekman layer. The case study here demonstrates the unique value of an integrated coastal ocean observing system in studying harmful algal blooms (HABs).     

Vertical migration of Karenia brevis in the northeastern Gulf of Mexico observed from glider measurements

The toxic marine dinoflagellate, Karenia brevis (the species responsible for most of red tides or harmful algal blooms in the Gulf of Mexico), is known to be able to swim vertically to adapt to the light and nutrient environments, nearly all such observations have been made through controlled experiments using cultures. Here, using continuous 3-dimensional measurements by an ocean glider across a K. brevis bloom in the northeastern Gulf of Mexico between 1 and 8 August 2014, we show the vertical migration behavior of K. brevis. Within the bloom where K. brevis concentration is between 100,000 and 1,000,000 cells L⁻¹, the stratified water shows a two-layer system with the depth of pycnocline ranging between 14–20 m and salinity and temperature in the surface layer being <34.8 and >28 °C, respectively. The bottom layer shows the salinity of >36 and temperature of <26 °C. The low salinity is apparently due to coastal runoff, as the top layer also shows high amount of colored dissolved organic matter (CDOM). Within the top layer, chlorophyll-a fluorescence shows clear diel changes in the vertical structure, an indication of K. brevis vertical migration at a mean speed of 0.5–1 m h⁻¹. The upward migration appears to start at sunrise at a depth of 8–10 m, while the downward migration appears to start at sunset (or when surface light approaches 0) at a depth of ∼2 m. These vertical migrations are believed to be a result of the need of K. brevis cells for light and nutrients in a stable, stratified, and CDOM-rich environment.     

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.     

Influence of daylight surface aggregation behavior on nutrient cycling during a Karenia brevis (Davis) G. Hansen & Ø. Moestrup bloom: Migration to the surface as a nutrient acquisition strategy

The toxic HAB dinoflagellate Karenia brevis (Davis) G. Hansen & Ø. Moestrup (formerly Gymnodinium breve) exhibits a migratory pattern atypical of dinoflagellates: cells concentrate in a narrow (∼0–5 cm) band at the water surface during daylight hours due to phototactic and negative geotactic responses, then disperse downward at night via non-tactic, random swimming. The hypothesis that this daylight surface aggregation behavior significantly influences bacterial and algal productivity and nutrient cycling within blooms was tested during a large, high biomass (chlorophyll a >19 μg L⁻¹) K. brevis bloom in October of 2001 by examining the effects of this surface layer aggregation on inorganic and organic nutrient concentrations, cellular nitrogen uptake, primary and bacterial productivity and the stable isotopic signature (δ¹⁵N, δ¹³C) of particulate material. During daylight hours, concentrations of K. brevis and chlorophyll a in the 0–5 cm surface layer were enhanced by 131% (±241%) and 32.1% (±86.1%) respectively compared with an integrated water sample collection over a 0–1 m depth. Inorganic (NH₄, NO₃₊₂, PO₄, SiO₄) and organic (DOP, DON) nutrient concentrations were also elevated within the surface layer as was both bacterial and primary productivity. Uptake of nitrogen (NH₄⁺, NO₃⁻, urea, dissolved primary amines, glutamine and alanine) compounds by K. brevis was greatest in the surface layer for all compounds tested, with the greatest enhancement evident in urea uptake rates, from 0.08 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹ to 3.1 × 10⁻⁵ ng N K. brevis cell⁻¹ h⁻¹. These data suggests that this surface aggregation layer is not only an area of concentrated cells within K. brevis blooms, but also an area of increased biological activity and nutrient cycling, especially of nitrogen. Additionally, the classic dinoflagellate migration paradigm of a downward migration for access to elevated NO₃⁻ concentrations during the dark period may not apply to certain dinoflagellates such as K. brevis in oligotrophic nearshore areas with no significant nitricline. For these dinoflagellates, concentration within a narrow surface layer in blooms during daylight hours may enhance nutrient supply through biological cycling and photochemical nutrient regeneration.     

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.     

Harmful algal bloom (HAB) in the East Sea identified by the Geostationary Ocean Color Imager (GOCI)

Harmful Cochlodinium polykrikoides blooms have frequently appeared and caused fatal harm to aquaculture in Korean coastal waters since 1995. We investigated the applicability of GOCI, the world's first Geostationary Ocean Color Imager, in monitoring the distribution and temporal movement of a harmful algal bloom (HAB) that was discovered in the East Sea near the Korean peninsula in August 2013. We identified the existence of C. polykrikoides at a maximum cell abundance of over 6000 cells/mL and a chlorophyll a concentration of over 400 mg/m³. In areas of C. polykrikoides blooms, GOCI remote sensing reflectance (R_rs) spectra demonstrated the typical radiometric features of a HAB, and from the diurnal variations using GOCI-derived chlorophyll concentration images, we were able to identify the vertical migration of the red tide species. We also found that the formation and propagation of the HAB had relations with cold water mass in the coastal region. GOCI can be effectively applied to the monitoring of short-term and long-term movements of red tides.     

Nitrogen uptake kinetics in field populations and cultured strains of Karenia brevis

This study represents the most comprehensive assessment of kinetic parameters for Karenia brevis to date as it encompasses natural populations sampled during three different bloom years in addition to cultured strains under controlled conditions. Nitrogen (N) uptake kinetics for ammonium (NH₄⁺), nitrate (NO₃⁻), urea, an amino acid mixture, individual amino acids (glutamate and alanine), and humic substrates were examined for the toxic red tide dinoflagellate, K. brevis, during short term incubations (0.5–1 h) using ¹⁵N tracer techniques. Experiments were conducted using natural populations collected during extensive blooms along the West Florida Shelf in October 2001, 2002, and 2007, and in cultured strains (CCFWC 251 and CCFWC 267) obtained from the Florida Fish and Wildlife Institute culture collection. Kinetic parameters for the maximum uptake velocity (V_max), half-saturation concentration (K_s), and the affinity constant (α) were determined. The affinity constant is considered a more accurate indicator of substrate affinity at low concentrations. K. brevis took up all organic substrates tested, including N derived from humic substances. Uptake rates of the amino acid mixture and some NO₃⁻ incubations did not saturate even at the highest substrate additions (50–200 μmol N L⁻¹). Based upon the calculated α values, the greatest substrate preference was for NH₄⁺ followed by NO₃⁻ ≥ urea, humic compounds and amino acids. The ability of K. brevis to utilize a variety of inorganic and organic substrates likely helps it flourish under a wide range of nutrient conditions from bloom initiation in oligotrophic waters offshore to bloom maintenance near shore where ambient nutrient concentrations may be orders of magnitude greater.     

Zooplankton community composition and copepod grazing on the West Florida Shelf in relation to blooms of Karenia brevis

Blooms of the toxin producing dinoflagellate Karenia brevis occur routinely on the West Florida Shelf of the Gulf of Mexico. Nutrient supplies are thought to play a large role in the formation and maintenance of these blooms. The role of top-down control has been less well studied, but grazing, or the lack thereof, on these toxic species may also enhance the formation of large biomass blooms in this region. Zooplankton community structure and copepod species composition were analyzed from samples collected on the West Florida Shelf (WFS) during a NOAA funded ECOHAB regional Karenia Nutrient Dynamics project during October 2007–2010. In 2008 there was no statistical difference in the abundance of zooplankton at bloom and non-bloom stations, however in 2009 there was a statistically significant difference (p < 0.05) between the abundance of zooplankton at stations with Karenia present. To investigate copepod ingestion rates in relation to K. brevis, shipboard and laboratory experiments of the single label method of ¹⁴C labeled phytoplankton culture, and time course ingestion experiments with isolated copepods were performed. Calculated ingestion rates suggest that the copepod species Centropages velificatus, and Acartia tonsa ingested K. brevis, however rates were variable among collection sites and K. brevis strains. Parvocalanus crassirostris did not ingest K. brevis in any of the experiments.     

Empirical models of toxigenic Pseudo-nitzschia blooms: Potential use as a remote detection tool in the Santa Barbara Channel

The Santa Barbara Channel, CA is a highly productive region where wind-driven upwelling and mesoscale eddies are important processes driving phytoplankton blooms. In recent years, the spring bloom has been dominated by the neurotoxin-producing diatom, Pseudo-nitzschia spp. In this paper, we relate a 1.5-year time series of Pseudo-nitzschia spp. abundance and domoic acid concentration to physical, chemical, and biological data to better understand the mechanisms controlling local Pseudo-nitzschia spp. bloom dynamics. The data were used to define the ranges of environmental conditions associated with Pseudo-nitzschia spp. bloom development in the Santa Barbara Channel. The time series captured three large toxic events (max. particulate domoic acid concentration, pDA ～6000 ng L^?1; max. cellular domoic acid concentrations, cDA ～88 pg cell^?1) in the springs of 2005–2006 and summer 2005 corresponding to bloom-level Pseudo-nitzschia spp. abundance (>5.0 × 10⁴ cells L^?1). In general, large increases in Pseudo-nitzschia spp. abundance were accompanied by increases in cDA levels, and cDA peaks preceded pDA peaks by at least one month in both the springs of 2005 and 2006. Statistical models incorporating satellite ocean color (MODIS-Aqua and SeaWiFS) and sea surface temperature (AVHRR) data were created to determine the probability that a remotely sensed phytoplankton bloom contains a significant population of toxic Pseudo-nitzschia spp. Models correctly estimate 98% of toxic bloom situations, with a 7–29% rate of false positive identification. Conditions most associated with high cDA levels are low sea surface temperature, high salinity, increased absorption by cDOM (412 nm), increased reflectance at 510/555 nm, and decreased particulate absorption at 510 nm. Future efforts to merge satellite and regionally downscaled forecasting products with these habitat models will help assess bloom forecasting capabilities in the central CA region and any potential connections to large-scale climate modes.     

Comparative diel oxygen cycles preceding and during a Karenia bloom in Sarasota Bay,Florida, USA

The diel change in dissolved oxygen concentrations were recorded with an automated incubator containing a pulsed oxygen sensor in Sarasota Bay, Florida. The deployments occurred during a ‘pre-bloom’ period in May to June 2006, and during a harmful algal bloom dominated by Karenia brevis in September 2006. The diurnal (daylight) increase in dissolved oxygen concentrations varied from 16 to 104 μmol O₂ l⁻¹ with the corresponding nocturnal decrease in oxygen varying from 16 to 77 μmol O₂ l⁻¹. Nocturnal respiration consumed 42–113% of the diurnal net oxygen production with the minimum and maximum during the pre-bloom period. Hourly production rates closely followed fluctuations in irradiance with maximum rates in the late morning. Hourly oxygen utilization rates (community respiration) at night were highest during the first few hours after sunset.     

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.     

Brevetoxin persistence in sediments and seagrass epiphytes of east Florida coastal waters

A bloom of Karenia brevis Davis developed in September 2007 near Jacksonville, Florida and subsequently progressed south through east Florida coastal waters and the Atlantic Intracoastal Waterway (ICW). Maximum cell abundances exceeded 10⁶ cells L⁻¹ through October in the northern ICW between Jacksonville and the Indian River Lagoon. The bloom progressed further south during November, and terminated in December 2007 at densities of 10⁴ cells L⁻¹ in the ICW south of Jupiter Inlet, Florida. Brevetoxins were subsequently sampled in sediments and seagrass epiphytes in July and August 2008 in the ICW. Sediment brevetoxins occurred at concentrations of 11–15 ng PbTx-3 equivalents (g dry wt sediment)⁻¹ in three of five basins in the northern ICW during summer 2008. Seagrass beds occur south of the Mosquito Lagoon in the ICW. Brevetoxins were detected in six of the nine seagrass beds sampled between the Mosquito Lagoon and Jupiter Inlet at concentrations of 6–18 ng (g dry wt epiphytes)⁻¹. The highest brevetoxins concentrations were found in sediments near Patrick Air Force Base at 89 ng (g dry wt sediment)⁻¹. In general, brevetoxins occurred in either seagrass epiphytes or sediments. Blades of the resident seagrass species have a maximum life span of less than six months, so it is postulated that brevetoxins could be transferred between epibenthic communities of individual blades in seagrass beds. The occurrence of brevetoxins in east Florida coast sediments and seagrass epiphytes up to eight months after bloom termination supports observations from the Florida west coast that brevetoxins can persist in marine ecosystems in the absence of sustained blooms. Furthermore, our observations show that brevetoxins can persist in sediments where seagrass communities are absent.     

Effects of nutrient-limiting supply ratios on toxin content of Karenia brevis grown in continuous culture

Toxins produced as secondary metabolites can play important roles in phytoplankton communities and contribute to the ecological success of harmful algal bloom (HAB) taxa. Toxin composition and content in phytoplankton are affected by a suite of environmental factors, including nutrient availability. Changes in nutrient availability can increase or decrease toxin content and alter toxin composition, depending on toxin stoichiometry and the mechanisms by which nutrient limitation affects toxin production. The studies that have assessed the effects of nutrient availability on brevetoxin content of the HAB species Karenia brevis have reported contradictory results, although there is growing support that nutrient limitation increases brevetoxin content. In this study, we assessed the effects of decreased nitrogen (N) and phosphorus (P) availability on brevetoxin content and composition of K. brevis grown in chemostats at steady state by altering the nutrient supply ratios of incoming media from the Redfield Ratio. Overall, brevetoxin content was greatest in cultures grown at the lowest rate, regardless of the nutrient supply ratio (i.e., under both Redfield and N-limiting supply ratios). Compared to cultures grown at 0.2 d⁻¹, cultures grown at 0.1 d⁻¹ exhibited 5-fold increases in intracellular toxin content. In contrast, at constant growth rates, N-limiting supply ratios decreased intracellular brevetoxin content by approximately one-third, although this result was significant only in cultures growing at the fastest rate of 0.23 d⁻¹. P-limiting supply ratios had no effect on brevetoxin content or composition. In addition, when cultures grown at rates of 0.2 d⁻¹ were supplied with balanced/Redfield N:P supply ratios, but different absolute nutrient concentrations, toxin content was greater under greater nutrient concentrations. These findings suggest that when growth rate is not nutrient limited, there is a positive relationship between nutrient availability and brevetoxin content. This work contributes to previous studies by demonstrating strong growth rates effects on brevetoxin content and that growth rate and nutrient availability can independently or together affect toxin content of K. brevis. Moreover, our work underscores the value of the chemostat as a tool to elucidate the mechanisms by which nutrient availability and growth rate affect toxin production and content of HAB species.     

The applicability of empirical vegetation indices for determining leaf chlorophyll content over different leaf and canopy structures

Retrieving leaf chlorophyll content at a range of spatio-temporal scales is central to monitoring vegetation productivity, identifying physiological stress and managing biological resources. However, estimating leaf chlorophyll over broad spatial extents using ground-based traditional methods is time and resource heavy. Satellite-derived spectral vegetation indices (VIs) are commonly used to estimate leaf chlorophyll content, however they are often developed and tested on broadleaf species. Relatively little research has assessed VIs for different leaf structures, particularly needle leaves which represent a large component of boreal forest and significant global ecosystems. This study tested the performance of 47 published VIs for estimating foliar chlorophyll content from different leaf and canopy structures (broadleaf and needle). Coniferous and deciduous sites were selected in Ontario, Canada, representing different dominant vegetation species (Picea mariana and Acer saccharum) and a variety of canopy structures. Leaf reflectance data was collected using an ASD Fieldspec Pro spectroradiometer (400–2500 nm) for over 300 leaf samples. Canopy reflectance data was acquired from the medium resolution imaging spectrometer (MERIS). At the canopy level, with both leaf types combined, the DD-index showed the strongest relationship with leaf chlorophyll (R² = 0.78; RMSE = 3.56 μg/cm²), despite differences in leaf structure. For needleleaf trees alone the relationship with the top VI was weaker (D_[red], R² = 0.71; RMSE = 2.32 μg/cm²). A sensitivity study using simulated VIs from physically-modelled leaf (PROSPECT) and canopy (4-Scale) reflectance was performed in order to further investigate these results and assess the impacts of different background types and leaf area index on the VIs’ performance. At the leaf level, the MNDVI8 index showed a strong linearity to changing chlorophyll and negligible difference to leaf structure/type. At canopy level, the best performing VIs were relatively consistent where LAI ≥ 4, but responded strongly to differences in background at low canopy coverage (LAI = 2). This research provides comprehensive assessments for the use of spectral indices in retrieval of spatially-continuous leaf chlorophyll content at the leaf (MTCI: R² = 0.72; p < 0.001) and canopy (DD: R² = 0.78; p < 0.001) level for resource management over different spatial and temporal scales.     

Contribution of diazotrophy to nitrogen inputs supporting Karenia brevis blooms in the Gulf of Mexico

Blooms of Karenia brevis plague the West Florida Shelf (WFS) region in the Gulf of Mexico (GOM) where they exert harmful effects on aquatic biota and humans. Because productivity on the WFS is N limited, new N inputs into the region are thought to trigger blooms of K. brevis. Here we examine the potential for new N inputs via N₂ fixation by Trichodesmium and other diazotrophic plankton to contribute to the N demand of K. brevis. Because of possible methodological biases, we also compared N₂ fixation rates by cultured Trichodesmium using the ¹⁵N₂ bubble addition method and the ¹⁵N₂ saturated seawater. Both methods yielded identical results in 12 and 24 h incubations; however, there was more variability in rate estimates made using the bubble addition method. Pelagic N₂ fixation rates by other planktonic diazotrophs ranged from 0 to 13.6 nmol N L⁻¹ d⁻¹, comparable to or higher than rates observed in oligotrophic gyres. These rates should be considered conservative estimates because they were made using the bubble addition method. Integrating over our study area, we estimate that new inputs of N to the WFS via N₂ fixation are on the order of 0.011 Tmol N annually. Further, we measured directly the trophic transfer of recently fixed N₂ to co-occurring plankton that included K. brevis and found that up to 47% of N₂ fixed was transferred to non-diazotrophic plankton even in short (<6 h) incubations where N₂ fixation was likely underestimated.     

Brevenal,a brevetoxin antagonist from Karenia brevis,binds to a previously unreported site on mammalian sodium channels

Brevetoxins are a family of ladder-frame polyether toxins produced by the marine dinoflagellate Karenia brevis. During blooms of K. brevis, inhalation of brevetoxins aerosolized by wind and wave action can lead to asthma-like symptoms in persons at the beach. Consumption of either shellfish or finfish contaminated by K. brevis blooms can lead to the development of neurotoxic shellfish poisoning. The toxic effects of brevetoxins are due to binding at a defined site on, and subsequent activation of, voltage-sensitive sodium channels (VSSCs) in cell membranes (site 5). In addition to brevetoxins, K. brevis produces several other ladder-frame compounds. One of these compounds, brevenal, has been shown to antagonize the effects of brevetoxin. In an effort to further characterize the effects of brevenal, a radioactive analog ([³H]-brevenol) was produced by reducing the terminal aldehyde moiety of brevenal to an alcohol using tritiated sodium borohydride. A K_D of 67 nM and B_max of 7.1 pmol/mg protein were obtained for [³H]-brevenol in rat brain synaptosomes, suggesting a 1:1 matching with VSSCs. Brevenal and brevenol competed for [³H]-brevenol binding with K_i values of 75 nM and 56 nM, respectively. However, although both brevenal and brevenol inhibited brevetoxin binding, brevetoxin was completely ineffective at competition for [³H]-brevenol binding. After examining other site-specific compounds, it was determined that [³H]-brevenol binds to a site that is distinct from the other known sites on the sodium channel, including the brevetoxin site, (site 5) although some interaction with site 5 is apparent.     

Distribution and dynamics of two species of Dinophyceae producing high biomass blooms over the French Atlantic Shelf

The frequency and distribution of high biomass blooms produced by two dinoflagellate species were analysed along the French continental shelf from 1998 to 2012. Two species were specifically studied: Karenia mikimotoi and Lepidodinium chlorophorum. Based on remote-sensing reflectances at six channels (410, 430, 480, 530, 550 and 670 nm), satellite indices were created to discriminate the species forming the blooms. A comparison with observations showed that the identification was good for both species in spite of a lower specificity for L. chlorophorum. The overall analysis of the satellite indices, in association with some monitoring data and cruise observations, highlights the regularity of these events and their extent on the continental shelf. L. chlorophorum blooms may occur all along the South Coast of Brittany. All the coastal areas under the influence of river plumes and the stratified northern shelf area of the Western English Channel appear to be areas of bloom events for both species. These two species are likely to be in competitive exclusion as they share the same spatial distribution and the timing of their bloom is very close. Finally, due to the scarcity of off-shore observations, these satellite indices provide useful information regarding HABs management and the development of a warning system along the French coast.     

Molecular ecological responses of the dinoflagellate Karenia mikimotoi to phosphate stress

Karenia mikimotoi is a toxic, widespread dinoflagellate which could produce hemolytic toxins and ichthyotoxins affecting fisheries within the area of its bloom. Previous ecophysiological studies indicated that the enhance of environmental phosphate concentration could promote the growth of K. mikimotoi. Intrinsic mechanisms regarding the effects of external phosphate on its photosynthesis, cell cycle succession and differential proteins’ expressions are still unknown. K. mikimotoi was cultured in phosphate-deprived medium, while the culture in f/2 medium (Guillard, 1975) was introduced as phosphate-sufficient control experiment. Cell counts and phosphate concentration detection were performed every other day. Flowcytometry was applied to measure cell cycle succession and chlorophyll fluorescence intensity fluctuation. Differential proteomics expression was examined by SDS-PAGE tandem LTQ Orbitrap MS/MS spectrometry. Functions of each differential protein were searched within NCBInr protein database and Swissprot database. Our study demonstrated that phosphate stress inhibited growth and cell cycle succession of K. mikimotoi remarkably (p < 0.01). Algal chlorophyll fluorescence intensity was significantly affected by phosphate deprivation (p < 0.05). 11 species of differential proteins were detected only in phosphate-limited culture sample which related to stress signal transduction, vacuolar phosphate release, phospholipid degradation, organic acid synthesis and phagotrophy. 4 kinds of differential proteins were identified only in f/2 medium culture sample which referred to cell proliferation, glycolysis, SAM cycle and polyamine production. Based on analysis of differential proteomic functional annotation, we hypothesized proteomic response mechanism of K. mikimotoi to phosphate stress. Molecular biological responses of dinoflagellate K. mikimotoi to phosphate stress was explored.     

Effects of phytoplankton species composition on absorption spectra and modeled hyperspectral reflectance

Understanding the spectral characteristics of remotely-sensed reflectance by different phytoplankton species can assist in the development of algorithms to identify various algal groups using satellite ocean color remote sensing. One of the main challenges is to separate the effect of species composition on the reflectance spectrum from other factors such as pigment concentration and particle size structure. Measuring the absorption spectra of nine different cultured algae, and estimating the reflectance of the different species, provides a useful approach to study the effects of species composition on the bio-optical properties. The results show that the absorption spectra of different species exhibit different spectral characteristics and that species composition can significantly change the absorption characteristics at four main peaks (438, 536, 600 and 650 nm). A ‘distance angle index’ was used to compare different phytoplankton species. Results indicate that this index can be used to identify species from the absorption spectra, using a database of standard absorption spectra of known species as reference. By taking into account the role of species composition in the phytoplankton absorption model, the performance of the model can be improved by up to 5%. A reflectance-species model is developed to estimate the remotely-sensed reflectance from the absorption spectra, and the reflectance of different phytoplankton species at the same chlorophyll-a concentration is compared, to understand effects of species composition on the reflectance spectra. Different phytoplankton species can cause up to 33% difference in the modeled reflectance at short wavelengths under the condition of the same chlorophyll-a concentration, and variations in the reflectance spectrum correspond to the colors of the algae. The standard deviation of the reflectance among different species shows that the variations from 400 to 450 nm are sensitive to species composition at low chlorophyll-a concentrations, whereas variations in the 510 to 550 nm range are more sensitive under high chlorophyll-a concentrations. For this reason, the green bands may be more suitable for estimating species composition from hyperspectral satellite data during bloom conditions, whereas the blue bands may be more helpful in detection of species under low chlorophyll-a concentrations. In this theoretical approach, variations in reflectance at the same chlorophyll-a concentration can be used to identify phytoplankton species. Another approach to identify phytoplankton species from remotely-sensed hyperspectral reflectance measurements would be to derive the absorption spectra of phytoplankton from the reflectance measurements, and compare these with a standard database of absorption spectra.