Neurological illnesses associated with Florida red tide (Karenia brevis) blooms

Human respiratory and gastrointestinal illnesses can result from exposures to brevetoxins originating from coastal Florida red tide blooms, comprising the marine alga Karenia brevis (K. brevis). Only limited research on the extent of human health risks and illness costs due to K. brevis blooms has been undertaken to date. Because brevetoxins are known neurotoxins that are able to cross the blood-brain barrier, it is possible that exposure to brevetoxins may be associated with neurological illnesses. This study explored whether K. brevis blooms may be associated with increases in the numbers of emergency department visits for neurological illness. An exposure-response framework was applied to test the effects of K. brevis blooms on human health, using secondary data from diverse sources. After controlling for resident population, seasonal and annual effects, significant increases in emergency department visits were found specifically for headache (ICD-9 784.0) as a primary diagnosis during proximate coastal K. brevis blooms. In particular, an increased risk for older residents (≥55 years) was identified in the coastal communities of six southwest Florida counties during K. brevis bloom events. The incidence of headache associated with K. brevis blooms showed a small but increasing association with K. brevis cell densities. Rough estimates of the costs of this illness were developed for hypothetical bloom occurrences.     

Removal of harmful algal cells (Karenia brevis) and toxins from seawater culture by clay flocculation

Harmful algal blooms (HABs) occur worldwide causing serious threat to marine life, and to public health through seafood-borne illnesses and exposure to toxin-containing marine aerosol. This study was undertaken to assess the ability of phosphatic clay to remove the toxic dinoflagellate, Karenia brevis, and the potent neurotoxins (brevetoxins) produced by this species. Results showed that the addition of an aqueous slurry of 0.75 g (dry weight) clay to 3 l of K. brevis culture, containing 5×10⁶ and 10×10⁶ cells/l, removed 97±4% of brevetoxins from the water column within 4 h after the addition of clay. Clay flocculation of extra-cellular brevetoxins, released from cells ruptured (lyzed) by ultrasonication, removed 70±10% of the toxins. Addition of the chemical flocculant, polyaluminum chloride (PAC), removed all of the extra-cellular toxins. A 14 day study was undertaken to observe the fate of brevetoxins associated with clay flocculation of viable K. brevis cells. At 24 h following the clay addition, 90±18% of the toxins were removed from the water column, along with 85±4% of the cells. The toxin content of clay diminished from 208±13 μg at Day 1, to 121±21 μg at Day 14, indicating that the phosphatic clay retained about 58% of the toxins throughout the 14-day period. These studies showed the utility of natural clay as a means of reducing adverse effects from HABs, including removal of dissolved toxins, in the water column, although considerable work clearly remains before this approach can be used on natural blooms in open waters.     

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.     

Karenia brevis monitoring,management, and mitigation for Florida molluscan shellfish harvesting areas

The purpose of this paper is to describe the State of Florida Karenia brevis monitoring, management and mitigation procedures for the harvesting of safe molluscan shellfish. Monitoring and management to prevent public health impacts of due to brevetoxins from K. brevis in shellfish has worked successfully for over forty years. Over the past forty years there have been no reports of human illness resulting from the consumption of commercial or recreational harvest harvested of shellfish from Florida waters that were classified as open. Therefore, this monitoring and management program has been a highly successful public health program. In addition, opportunities exist for mitigation which may allow for limited shellfish harvesting and safe consumption during blooms. The key elements of the Florida program are provided as a “model” or “ideal” monitoring, management, and mitigation program.     

Tracking losses of brevetoxins on exposure to phytoplankton competitors: Ecological impacts

The frequent occurrence of devastating blooms of the harmful dinoflagellate Karenia brevis in the Gulf of Mexico has motivated research into bloom dynamics and potential mitigation strategies. The use of competing phytoplankton to lower waterborne concentrations of the most abundant and toxic brevetoxins produced during these blooms has been proposed. However the ecological impacts of using such biocontrol agents have not been addressed. This study investigated the impact on marine invertebrates of lowered brevetoxin concentrations due to the presence of competing phytoplankton. Even at low brevetoxin concentrations, the presence of the common diatom Skeletonema grethae ameliorated harmful toxic effects of brevetoxins upon the brine shrimp, Artemia salina, and reduced the incidence of negative physiological and morphological responses of the sea anemone Aiptasia pallida. In addition, brevetoxin biotransformation products formed by competing phytoplankton appear to be non-toxic or do not trigger the same physiological responses as brevetoxins in the model organisms used. These findings may impact the interpretation of ecotoxicological data gathered during bloom events, since the presence of phytoplankton competitors in Karenia blooms is likely to reduce the harmful effects seen on many marine invertebrates.     

Loss of waterborne brevetoxins from exposure to phytoplankton competitors

We tested whether interactions among phytoplankton competitors affect toxin dynamics involving the red tide dinoflagellate Karenia brevis, whose brevetoxins incapacitate and kill coastal wildlife. The addition of a live diatom, Skeletonema costatum, led to decreased concentrations of brevetoxin B (PbTx-2) associated with K. brevis cells in co-culturing experiments and with two of three natural bloom samples containing K. brevis. Similar decreases in PbTx-2 concentration, but not PbTx-3 concentration, occurred when a mixture of brevetoxins (without live K. brevis cells) was exposed to S. costatum, indicating that S. costatum metabolizes waterborne PbTx-2. Liquid chromatography–mass spectrometry (LC–MS) and ELISA analyses indicated that PbTx-2 is probably not transformed into other brevetoxins or into known brevetoxin metabolites, and instead is biotransformed by a previously unrecognized mechanism. Four different S. costatum strains from around the world caused similar loss of PbTx-2, suggesting that evolutionary experience with K. brevis is not a pre-requisite for the ability to metabolize PbTx-2. Additionally, phytoplankton-associated bacteria were found to play no role in the loss of PbTx-2, as bacteria-free S. costatum strains metabolized PbTx-2. Finally, loss of waterborne PbTx-2 caused by exposure to a dinoflagellate, a cryptophyte, and two additional diatom species indicates that this phenomenon is widespread among phytoplankton. Our results unexpectedly suggest that competing phytoplankton species present during K. brevis blooms, and possibly other red tides, could mediate bloom toxicity and therefore ecosystem-level consequences of red tides.     

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.     

Karenia brevis red tides,brevetoxins in the food web,and impacts on natural resources: Decadal advancements

As recently as a decade ago, Karenia brevis red tides and their effects on animal resources in the Gulf of Mexico were principally perceived as acute blooms that caused massive fish kills. Although occasional mortalities of higher vertebrates were documented, it has only been in the past decade that conclusive evidence has unequivocally demonstrated that red tides and their brevetoxins are lethal to these organisms. Brevetoxins can be transferred through the food chain and are accumulated in or transferred by biota at many trophic levels. The trophic transfer of brevetoxins in the food web is a complex phenomenon, one that is far more complicated than originally conceived. Unexplained fish kills and other animal mortalities in areas where red tide is endemic are being increasingly linked with post-bloom exposures of biota to brevetoxins. Mass mortality events of endangered Florida manatees (Trichechus manatus latirostris) follow a consistent spatial and temporal pattern, occurring primarily in the spring in southwestern Florida. Persistent blooms can also cause a cascade of environmental changes, affecting the ecosystem and causing widespread die-offs of benthic communities. Ongoing fish kills from sustained blooms can lead to short-term declines in local populations. Although animal populations in areas where red tide is endemic are unquestionably at risk, it remains to be determined to what extent populations can continue to recover from these sustained effects.     

Florida red tide and brevetoxins: Association and exposure in live resident bottlenose dolphins (Tursiops truncatus) in the eastern Gulf of Mexico,U.S.A.

Bottlenose dolphins (Tursiops truncatus) along the Gulf of Mexico are frequently exposed to blooms of the toxic alga, Karenia brevis, and brevetoxins associated with these blooms have been implicated in several dolphin mortality events. Studies on brevetoxin accumulation in dolphins have typically focused on analyses of carcasses from large‐scale die‐offs; however, data are scarce for brevetoxin loads in live individuals frequently exposed to K. brevis blooms. This study investigated in vivo brevetoxin exposure in free‐ranging bottlenose dolphins resident to Sarasota Bay, Florida, utilizing samples collected during health assessments performed during multiple K. brevis blooms occurring from 2003 to 2005. Brevetoxins were detected by ELISA and LC‐MS in 63% of bottlenose dolphins sampled (n= 30) concurrently with a K. brevis bloom. Brevetoxins were present in urine and gastric samples at concentrations ranging from 2 to 9 ng PbTx‐3 eq/g, and in feces at concentrations ranging from 45 to 231 ng PbTx‐3 eq/g. Samples from individuals (n= 12) sampled during nonbloom conditions (≤1,000 cells/L) were negative for brevetoxin activity. Brevetoxin accumulation data from this study complement dolphin carcass and prey fish data from the same study area, and aid in evaluating impacts of harmful algal blooms on sentinel marine animal species along the west Florida coast.     

Review of Florida Red Tide and Human Health Effects

This paper reviews the literature describing research performed over the past decade on the known and possible exposures and human health effects associated with Florida red tides. These harmful algal blooms are caused by the dinoflagellate, Karenia brevis, and similar organisms, all of which produce a suite of natural toxins known as brevetoxins. Florida red tide research has benefited from a consistently funded, long term research program, that has allowed an interdisciplinary team of researchers to focus their attention on this specific environmental issue-one that is critically important to Gulf of Mexico and other coastal communities. This long-term interdisciplinary approach has allowed the team to engage the local community, identify measures to protect public health, take emerging technologies into the field, forge advances in natural products chemistry, and develop a valuable pharmaceutical product. The Review includes a brief discussion of the Florida red tide organisms and their toxins, and then focuses on the effects of these toxins on animals and humans, including how these effects predict what we might expect to see in exposed people.     

Bacteriocidal effects of brevetoxin on natural microbial communities

The sensitivity of bacteria to the marine neurotoxins, brevetoxins, produced by the dinoflagellate Karenia brevis and raphidophytes Chattonella spp. remains an open question. We investigated the bacteriocidal effects of brevetoxin (PbTx-2) on the abundance and community composition of natural microbial communities by adding it to microbes from three coastal marine locations that have varying degrees of historical brevetoxin exposure: (1) Great Bay, New Jersey, (2) Rehoboth Bay, Delaware and (3) Sarasota Bay, Florida. The populations with limited or no documented exposure were more susceptible to the effects of PbTx-2 than the Gulf of Mexico populations which are frequently exposed to brevetoxins. The community with no prior documented exposure to brevetoxins showed significant (p = 0.03) changes in bacterial abundance occurring with additions greater than 2.5 μg PbTx-2 L⁻¹. Brevetoxin concentrations during K. brevis blooms range from ∼2.5 to nearly 100 μg L⁻¹ with typical concentrations of ∼10–30 μg L⁻¹. In contrast to the unexposed populations, there was no significant decrease in bacterial cell number for the microbial community that was frequently exposed to brevetoxins, which implies variable sensitivity in natural communities. The diversity in the bacterial communities that were sensitive to PbTx-2 declined upon exposure. This suggests that the PbTx-2 was selecting for or against specific species. Mortality was much higher in the 200 μg PbTx-2 L⁻¹ treatment after 48 h and >37% of the species disappeared in the bacterial communities with no documented exposure. These results suggest that toxic red tides may play a role in structuring bacterial communities.     

Progress in developing a new detection method for the harmful algal bloom species,Karenia brevis,through multiwavelength spectroscopy

Multiwavelength spectroscopy is a rapid analytical technique that can be applied to detect, identify, and quantify microorganisms such as Karenia brevis, the species known for frequent red-tide blooms in Florida's coastal waters. This research will report on a model-based interpretation of UV–vis spectra of K. brevis. The spectroscopy models are based on light scattering and absorption theories, and the approximation of the frequency-dependant optical properties of the basic constituents of living organisms. Absorption and scattering properties of K. brevis, such as cell size/shape, internal structure, and chemical composition, are shown to predict the spectral features observed in the measured spectra. The parameters for the interpretation model were based upon both reported literature values, and experimental values obtained from live cultures and pigment standards. Measured and mathematically derived spectra were compared to determine the adequacy of the model, contribute new spectral information, and to establish the proposed spectral interpretation approach as a new detection method for K. brevis.     

Karenia brevis causes high mortality and impaired swimming behavior of Florida stone crab larvae

The dinoflagellate Karenia brevis causes harmful algal blooms commonly referred to as red tides that are prevalent along Florida’s gulf coast. Severe blooms often cause fish kills, turbid water, and hypoxic events all of which can negatively impact local fisheries. The stone crab, Menippe mercenaria, is a ˜$25 million per year fishery that occurs primarily along Florida’s gulf coast. On the west Florida shelf, red tides occur from fall through spring, although severe blooms can occur during the summer. During the summer, stone crabs are reproductive and release larvae that are transported offshore where K. brevis blooms originate. This study determined the effects of K. brevis exposure on the survivorship, vertical swimming behavior, and oxygen consumption of stage-1 larval stone crabs. Survivorship was determined by exposing larvae to high (> 1 × 10⁶ cells L⁻¹) and medium (˜1 × 10⁵ cells L⁻¹) K. brevis concentrations for 96-hrs and were compared to controls that had no algae present. Larval swimming behavior (i.e., geotaxis) and oxygen consumption were monitored after 6-hr exposure to K. brevis. After 96-hrs of exposure, mortality was 100% and 30% for larvae in the high and medium concentrations of K. brevis, respectively, relative to the control. Larval swimming behavior was reversed in the K. brevis treatment; however oxygen consumption rates did not differ among treatments. These results suggest that severe blooms during the summer may reduce larval supply and serve as a potential bottleneck for new individuals recruiting into the fishery in years following a K. brevis bloom.     

Sublethal effects of the toxic dinoflagellate Karenia brevis on marine copepod behavior

Apart from grazing interactions, little is known regarding thesublethal effects of Karenia brevis cells on copepod behavior.We conducted grazing and mortality experiments with K. breviscells and brevetoxins (PbTx-2), establishing routes of toxicityfor the copepods Acartia tonsa, Temora turbinata and Centropagestypicus. Subsequent behavioral experiments determined whethercopepod swimming and photobehavior, both behaviors involvedin predator avoidance, were impaired at sublethal K. brevisand PbTx-2 levels. Copepods variably grazed toxic K. brevisand non-toxic Prorocentrum minimum at bloom concentrations.Although copepods accumulated brevetoxins, significant mortalitywas only observed in T. turbinata at the highest test concentration(1 x 10⁷ K. brevis cells L^–1). Acartia tonsa exhibitedminimal sublethal behavioral effects. However, there were significanteffects on the swimming and photobehavior of T. turbinata andC. typicus at the lowest sublethal concentrations tested (0.15µg PbTx-2 L^–1, 1 x 10⁵ K. brevis cells L^–1).Although physiological incapacitation may have altered copepodbehavior, starvation likely played a major role as well. Thesedata suggest that sublethal effects of K. brevis and brevetoxinon copepod behavior occur and predicting the role of zooplanktongrazers in trophic transfer of algal toxins requires knowledgeof species-specific sublethal effects.     

Monitoring of the toxic dinoflagellate Karenia brevis using gyroxanthin-based detection methods

The threat to human health and fisheries resources due to blooms of the toxic dinoflagellate Karenia brevis has lead to widespread public concern and calls for continuous monitoring of coastal waters for this organism. Here, a rapid and sensitive photopigment-based monitoring approach is described that incorporates refinements to standard filtration and analytical methods. This method uses the biomarker pigment gyroxanthin-diester contained in cells of some gymnodiniod species including K. brevis. Investigations of the retention efficiencies of five filter types for gyroxanthin from natural blooms of K. brevis showed no significant differences between GF/F, GF/C, 934-AH, GF/A or GF/D filters. Retention efficiencies were generally greater than 98% of cells added, indicating that the larger nominal pore size filters may be used safely for sample collection, reducing overall filtration times for large volumes of water. Simulated bloom experiments using cultures of K. brevis added to unfiltered water from Galveston Bay showed that retention of gyroxanthin on GF/D filters was significantly lower than on other filter types. There were significant interactions (p < 0.01) between filter type and cell density for the variables gyroxanthin, gyroxanthin chl a^–1 and gyroxanthin cell^–1, suggesting that the performance of the different filter types was dependent on cell density. Retention efficiencies for the simulated blooms ranged between >99% of cells retained and <30% of cells retained (greatest losses were for the GF/D filters). Combined results of natural and simulated blooms indicated that GF/C, 934-AH or GF/A filters gave the best retention efficiency with the fastest filtration times. Sample processing times were also improved by modifying the flow gradients in an existing HPLC protocol allowing the analysis of 106 samples in 24 h. The resulting protocol is suitable for incorporation into routine water quality monitoring programs, and would greatly facilitate the early detection and tracking of K. brevis blooms in coastal waters.     

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.     

Increased Toxicity of Karenia brevis during Phosphate Limited Growth: Ecological and Evolutionary Implications

Karenia brevis is the dominant toxic red tide algal species in the Gulf of Mexico. It produces potent neurotoxins (brevetoxins [PbTxs]), which negatively impact human and animal health, local economies, and ecosystem function. Field measurements have shown that cellular brevetoxin contents vary from 1–68 pg/cell but the source of this variability is uncertain. Increases in cellular toxicity caused by nutrient-limitation and inter-strain differences have been observed in many algal species. This study examined the effect of P-limitation of growth rate on cellular toxin concentrations in five Karenia brevis strains from different geographic locations. Phosphorous was selected because of evidence for regional P-limitation of algal growth in the Gulf of Mexico. Depending on the isolate, P-limited cells had 2.3- to 7.3-fold higher PbTx per cell than P-replete cells. The percent of cellular carbon associated with brevetoxins (%C-PbTx) was ∼ 0.7 to 2.1% in P-replete cells, but increased to 1.6–5% under P-limitation. Because PbTxs are potent anti-grazing compounds, this increased investment in PbTxs should enhance cellular survival during periods of nutrient-limited growth. The %C-PbTx was inversely related to the specific growth rate in both the nutrient-replete and P-limited cultures of all strains. This inverse relationship is consistent with an evolutionary tradeoff between carbon investment in PbTxs and other grazing defenses, and C investment in growth and reproduction. In aquatic environments where nutrient supply and grazing pressure often vary on different temporal and spatial scales, this tradeoff would be selectively advantageous as it would result in increased net population growth rates. The variation in PbTx/cell values observed in this study can account for the range of values observed in the field, including the highest values, which are not observed under N-limitation. These results suggest P-limitation is an important factor regulating cellular toxicity and adverse impacts during at least some K. brevis blooms.     

NITROGEN LIMITATION INCREASES BREVETOXINS IN KARENIA BREVIS (DINOPHYCEAE): IMPLICATIONS FOR BLOOM TOXICITY1

Laboratory and field measurements of the toxin content in Karenia brevis cells vary by >4‐fold. These differences have been largely attributed to genotypic variations in toxin production among strains. We hypothesized that nutrient limitation of growth rate is equally or more important in controlling the toxicity of K. brevis, as has been documented for other toxic algae. To test this hypothesis, we measured cellular growth rate, chlorophyll a, cellular carbon and nitrogen, cell volume, and brevetoxins in four strains of K. brevis grown in nutrient‐replete and nitrogen (N)‐limited semi‐continuous cultures. N‐limitation resulted in reductions of chlorophyll a, growth rate, volume per cell and nirtogen:carbon (N:C) ratios as well as a two‐fold increase (1%–4% to 5%–9%) in the percentage of cellular carbon present as brevetoxins. The increase in cellular brevetoxin concentrations was consistent among genetically distinct strains. Normalizing brevetoxins to cellular volume instead of per cell eliminated much of the commonly reported toxin variability among strains. These results suggest that genetically linked differences in cellular volume may affect the toxin content of K. brevis cells as much or more than innate genotypic differences in cellular toxin content per unit of biomass. Our data suggest at least some of the >4‐fold difference in toxicity per cell reported from field studies can be explained by limitation by nitrogen or other nutrients and by differences in cell size. The observed increase in brevetoxins in nitrogen limited cells is consistent with the carbon:nutrient balance hypothesis for increases in toxins and other plant defenses under nutrient limitation.     

Historical perspective on Karenia brevis red tide research in the Gulf of Mexico

Research on Karenia brevis blooms in the Gulf of Mexico started with the 1946–1947 red tide along the Florida west coast. Early research was on the organism itself, its tolerances and requirements, and the environment in which it lived and grew. Control of blooms, as a management option, was pursued in the 1950s with little success. However, in the 1960s–1970s, new regulation of shellfish growing areas was a public health management success. Research on K. brevis blooms followed funding cycles and was sporadic until the late 1990s when the National Oceanic and Atmospheric Administration (NOAA) and the Environmental Protection Agency (EPA) funded the Ecology and Oceanography of Harmful Algal Blooms (ECOHAB) and NOAA Monitoring and Event Response of Harmful Algal Blooms (MERHAB) programs. These particular funding programs, augmented by State of Florida appropriations, provided the opportunity to study K. brevis blooms on different temporal-spatial scales and consequently advanced the science. This review looks at historical research results in the light of today's advances.