Search Results (35)

To identify differentially abundant polyketide synthases (PKSs) and to characterize the biochemical consequences of brevetoxin biosynthesis, bottom-up, TMT-based quantitative proteomics and redox proteomics were conducted to compare two strains of the Florida red tide dinoflagellate Karenia brevis, which differ significantly in their brevetoxin content. Forty-eight PKS enzymes potentially linked to brevetoxin production were identified, with thirty-eight showing up to 16-fold higher abundance in the high-toxin strain. A pronounced shift toward a more oxidized redox state was observed in this strain’s proteome. Notably, 25 antioxidant-related proteins were significantly elevated, including alternative oxidase (AOX), which increased by 17-fold. These results elucidate the cellular consequences of toxin biosynthesis in K. brevis, offer new leads for the study of brevetoxin biosynthesis, and suggest a novel red tide mitigation approach targeting high toxin-producing strains. Full article

The Programmable Hyperspectral Seawater Scanner (PHySS) represents a significant breakthrough in monitoring harmful algal blooms (HABs), specifically targeting the “Florida red tide” caused by Karenia brevis. By utilizing a Fourth-Derivative Spectral Similarity Index (SI), this study establishes a strong positive correlation between the SI and phytoplankton counts, underscoring the PHySS’s potential for early detection and effective management of HABs. Our findings suggest that the PHySS could act as a predictive tool, offering crucial lead time to mitigate the ecological and economic repercussions of blooms. However, this study also identifies certain limitations of the PHySS technology, such as its inability to differentiate among various phytoplankton species without additional physical verification of cell counts. This limitation highlights the need for a multifaceted approach to HAB management. Our research suggests that adopting a multi-modal monitoring strategy could lead to more sophisticated and effective methods for combating HABs, fostering an optimistic outlook for future advancements in this area. Full article

Blooms of the harmful dinoflagellate Karenia brevis on the West Florida Shelf (WFS), Gulf of Mexico, are hypothesized to initiate in association with the colonial cyanobacterium Trichodesmium spp. and benefit from dissolved organic nitrogen (DON) release derived from N₂-fixation by the cyanobacteria. Previous studies have detected DON release using direct experimental measurements, but there have been few studies that have followed nutrient release by in situ blooms of Trichodesmium and the associated plankton community. It was determined that long-term Trichodesmium spp. and Karenia brevis abundances on the WFS were related, following a 2-month lag. A separate Eulerian study of a Trichodesmium erythraeum bloom event was conducted over 9 days on the Great Barrier Reef. Concentrations of T. erythraeum increased over the course of the study, with coincident increases in dinoflagellate abundance and decreases in diatom abundance. Inside the bloom, concentrations of NH₄⁺, PO₄³⁻, and DON increased significantly. The copepod grazer Macrosetella gracilis also increased in abundance as T. erythraeum numbers increased, contributing to nutrient release. Copepod grazing rates were measured, and N release rates estimated. Together, these studies show that Trichodesmium blooms have consequences for dinoflagellate abundance at both seasonal and ephemeral scales via direct and indirect N release. Full article

Previous studies have revealed that mixotrophs can become more heterotrophic as the temperature rises, although these studies were primarily conducted under laboratory conditions with temperature-acclimated grazers. This study investigated the short-term thermal regulation of grazing and photosynthetic performance, measured in terms of the maximum relative electron transport rate (rETR_max), of natural Karenia brevis populations on cultured Synechococcus. Bloom waters were collected in Sarasota, Florida, during the fall of 2022. Synechococcus were inoculated into K. brevis bloom waters in varying ratios and incubated at an ambient temperature and an ambient temperature ±5 °C (19, 24, and 29 °C). In general, the grazing coefficient, clearance, and ingestion rates were higher in warmer waters, although ingestion rates were significantly regulated by the prey-to-grazer ratios and, to a lesser degree, by temperatures (22 to 204 Synechococcus K. brevis⁻¹ d⁻¹). Overall, the rETR_max of Synechococcus controls generally increased over time with a more substantial increase at warmer temperatures, but, in the presence of grazers, the rETR_max of Synechococcus did not increase, and, remarkably, even decreased in some cases. These findings suggest that grazing on Synechococcus could directly regulate Synechococcus concentrations and indirectly reduce the photosynthetic performance of prey. Furthermore, this study demonstrates that the thermal regulation of grazing and photosynthetic performance can occur on a short-term basis. Full article

Curcumin, a natural plant product, was investigated as a mitigation tool against Karenia brevis, the toxic dinoflagellate responsible for Florida red tides. A series of laboratory bench-top studies were conducted with additions of 0.1, 1, 2, 3, 5, 10, 20, 30, and 40 mg/L curcumin to K. brevis at an average of 1.0 × 10⁶ cells/L to determine the efficacy of curcumin against K. brevis cells and brevetoxins and to optimize treatment dosage. Treatment with 5 mg/L of curcumin reduced K. brevis cell abundance by 89% and total brevetoxins by 60% within 24 h. Lower concentrations of curcumin (0.1–3 mg/L) exhibited between a 2 and 45% reduction in K. brevis and a reduction in brevetoxins of between 2 and 44% within 24 h. At the highest curcumin doses, 30 and 40 mg/L, a 100% reduction in cell abundance was observed by 6 h, with reduction in total brevetoxins by at least 64% in 48 h. These results suggest that curcumin, used alone or potentially in combination with other technologies, is a promising K. brevis bloom mitigation option. Full article

This study investigated how nitrogen and dissolved organic matter (DOM) from stormwater runoff and rainfall support the growth of Karenia brevis and Pyrodinium bahamense. Excitation–emission matrix spectroscopy coupled with parallel factor analysis tracked changes in the optical properties of DOM in each bioassay, revealing greater reactivity of terrestrial humic-like DOM. Significant increases in cell yield and specific growth rates were observed upon additions of runoff for both species, with significant increases in specific growth rates upon the addition of a 2 in simulated rain event for P. bahamense only. By hour 48, 100% of the dissolved organic nitrogen (DON) in each treatment was utilized by P. bahamense, and by hour 72, over 50% of the DON was utilized by K. brevis. The percentage of bioavailable dissolved organic carbon (DOC) was greater for P. bahamense compared to K. brevis, suggesting a greater affinity for DOC compounds by P. bahamense. However, the bioavailability of DOM for each species could be owed to distinct chemical characteristics of labile DOM conveyed from each site. This study demonstrates that stormwater runoff and rainfall are both sources of labile DOM and DON for K. brevis and P. bahamense, which has implications for blooms of these species in Tampa Bay waters. Full article

This review summarizes the current understanding of how brevetoxins, produced by Karenia brevis during harmful algal blooms, impact sea turtle health. Sea turtles may be exposed to brevetoxins through ingestion, inhalation, maternal transfer, and potentially absorption through the skin. Brevetoxins bind to voltage-gated sodium channels in the central nervous system, disrupting cellular function and inducing neurological symptoms in affected sea turtles. Moreover, the current evidence suggests a broader and longer-term impact on sea turtle health beyond what is seen during stranding events. Diagnosis relies on the detection of brevetoxins in tissues and plasma from stranded turtles. The current treatment of choice, intravenous lipid emulsion therapy, may rapidly reduce symptoms and brevetoxin concentrations, improving survival rates. Monitoring, prevention, and control strategies for harmful algal blooms are discussed. However, as the frequency and severity of blooms are expected to increase due to climate change and increased environmental pollution, continued research is needed to better understand the sublethal effects of brevetoxins on sea turtles and the impact on hatchlings, as well as the pharmacokinetic mechanisms underlying brevetoxicosis. Moreover, research into the optimization of treatments may help to protect endangered sea turtle populations in the face of this growing threat. Full article

Marine algae extracts are an important area of potential drug discovery; however, nearly all studies to date have used non-fluorescent-based methods to determine changes in target cell activity. Many of the most robust immunological and cellular analyses rely on fluorescent probes and readouts, which can be problematic when the algae extract is fluorescent itself. In this study, we identified the fluorescent spectrum of an isolated extract from the marine dinoflagellate Karenia brevis, which included two fluorescing components: chlorophyll α and pheophytin α. When excited at 405 nm and 664 nm, the extract emitted fluorescence at 676 nm and 696 nm, respectively. The extract and its fluorescing components, chlorophyll α and pheophytin α, entered phagocytic RAW 264.7 macrophages and non-phagocytic Vero kidney cells through distinct mechanisms. When incubated with the extract and its main components, both the RAW 264.7 macrophages and the Vero cells accumulated fluorescence as early as 30 min and continued through 48 h. Vero kidney cells accumulated the K. brevis fluorescent extract through a dynamin-independent and acidified endosomal-dependent mechanism. RAW 264.7 macrophages accumulated fluorescent extract through a dynamin-independent, acidified endosomal-independent mechanism, which supports accumulation through phagocytosis. Furthermore, RAW 264.7 macrophages downregulated cell-surface expression of CD206 in response to extract stimulation indicating activation of phagocytic responses and potential immunosuppression of these immune cells. This study represents the first characterization of the cellular update of K. brevis extracts in phagocytic versus non-phagocytic cells. The data suggest the importance of understanding cellular uptake of fluorescing algae extracts and their mechanism of action for future drug discovery efforts. Full article

Brevetoxins (BTXs) constitute a family of lipid-soluble toxic cyclic polyethers mainly produced by Karenia brevis, which is the main vector for a foodborne syndrome known as neurotoxic shellfish poisoning (NSP) in humans. To prevent health risks associated with the consumption of contaminated shellfish in France, the French Agency for Food, Environmental and Occupational Health & Safety (ANSES) recommended assessing the effects of BTXs via an acute oral toxicity study in rodents. Here, we investigated the effect of a single oral administration in both male and female mice with several doses of BTX-3 (100 to 1,500 µg kg⁻¹ bw) during a 48 h observation period in order to provide toxicity data to be used as a starting point for establishing an acute oral reference dose (ARfD). We monitored biological parameters and observed symptomatology, revealing different effects of this toxin depending on the sex. Females were more sensitive than males to the impact of BTX-3 at the lowest doses on weight loss. For both males and females, BTX-3 induced a rapid, transient and dose-dependent decrease in body temperature, and a transient dose-dependent reduced muscle activity. Males were more sensitive to BTX-3 than females with more frequent observations of failures in the grip test, convulsive jaw movements, and tremors. BTX-3’s impacts on symptomatology were rapid, appearing during the 2 h after administration, and were transient, disappearing 24 h after administration. The highest dose of BTX-3 administered in this study, 1,500 µg kg⁻¹ bw, was more toxic to males, leading to the euthanasia of three out of five males only 4 h after administration. BTX-3 had no effect on water intake, and affected neither the plasma chemistry parameters nor the organs’ weight. We identified potential points of departure that could be used to establish an ARfD (decrease in body weight, body temperature, and muscle activity). Full article

Climate change has been associated with intensified harmful algal blooms (HABs). Some harmful microalgae produce toxins that accumulate in food webs, adversely affecting the environment, public health and economy. Ocean acidification (OA) is a major consequence of high anthropogenic CO₂ emissions. The carbon chemistry and pH of aquatic ecosystems have been significantly altered as a result. The impacts of climate change on the metabolisms of microalgae, especially toxin biosynthesis, remain largely unknown. This hinders the optimization of HAB mitigation for changed climate conditions. To bridge this knowledge gap, previous studies on the effects of ocean acidification on toxin biosynthesis in microalgae were reviewed. There was no solid conclusion for the toxicity change of saxitoxin-producing dinoflagellates from the genus Alexandrium after high CO₂ treatment. Increased domoic acid content was observed in the diatom Pseudo-nitzschia. The brevetoxin content of Karenia brevis remained largely unchanged. The underlying regulatory mechanisms that account for the different toxicity levels observed have not been elucidated. Metabolic flux analysis is useful for investigating the carbon allocations of toxic microalgae under OA and revealing related metabolic pathways for toxin biosynthesis. Gaining knowledge of the responses of microalgae in high CO₂ conditions will allow the better risk assessment of HABs in the future. Full article

Harmful algal blooms (HABs) globally affect marine ecosystems and human health. Significant attention has been paid to understanding the initiation processes of HABs, while much less is known of the mechanisms causing cell demise and bloom decline. Recent evidence reveals that programmed cell death (PCD) can be a possible pathway for HAB termination. However, it is still not clear how PCD expression varies with the growth of marine phytoplankton. In order to characterize how susceptibility to stress-induced cell death changes with the growth of marine harmful algae, we examined the prevalence of PCD markers during 48 h after the addition of H₂O₂ in dinoflagellate Karenia brevis cultures at the early-log, mid-log and stationary phase. The results show that acute susceptibility to cell death (before time 0.5 h) increased as cultures aged, reflecting a chronological decrease in stress acclimation abilities. However, the youngest cultures showed strong PCD expression and the fastest overall rate of cell loss within the first 24 h, hypothesized to result from the cell-density-dependent H₂O₂ detoxifying process. This research highlights the existence of PCD in HAB species and that aging marine phytoplankton are more susceptible to exogenous stress, which agrees with previous observations of significant PCD during bloom decline. Full article

The harmful algal bloom species, Karenia brevis, forms annual, often intense blooms in the Gulf of Mexico, particularly along the west Florida shelf. Though the ability of K. brevis blooms to cause mass mortalities in juvenile fish are well documented, the direct effect of bloom concentrations on larval fish has not been studied extensively. To better understand the potential effect of K. brevis on larval fish survival, laboratory spawned red porgy (Pagrus pagrus) larvae from 4–26 days post-hatch were exposed to concentrations of K. brevis observed in the field for either 24 or 48 h. This species is representative of fish which spawn in regions of the Gulf of Mexico and whose larvae are epipelagic and may encounter K. brevis blooms. In this study, three different K. brevis strains varying in the amount of brevetoxin produced were tested. Larval survivorship was found to be inversely proportional to the amount of brevetoxin produced by each strain. The EC₅₀ value from the combined 24 h experiments was ~163,000 K. brevis cells L⁻¹, which corresponds to cell concentrations found in moderately dense blooms. Larval mortality also increased substantially in the 48 h versus 24 h exposure treatments. These findings indicate K. brevis blooms have the potential to contribute to natural mortality of fish larvae and further reduce inter-annual recruitment of fishery species whose stocks in the Gulf of Mexico may already be depleted. Full article

Karenia mikimotoi is a toxic bloom-forming dinoflagellate that sometimes co-blooms with Karenia brevis in the Gulf of Mexico, especially on the West Florida Shelf where strong vertical temperature gradients and rapid changes in nitrogen (N) can be found. Here, the short-term interactions of temperature, N form, and availability on photosynthesis–irradiance responses were examined using rapid light curves and PAM fluorometry in order to understand their interactions, and how they may affect photosynthetic yields. Cultures of K. mikimotoi were enriched with either nitrate (NO₃⁻), ammonium (NH₄⁺), or urea with varying amounts (1, 5, 10, 20, 50 µM-N) and then incubated at temperatures of 15, 20, 25, 30 °C for 1 h. At 15–25 °C, fluorescence parameters (Fv/Fm, rETR) when averaged for all N treatments were comparable. Within a given light intensity, increasing all forms of N concentrations generally led to higher photosynthetic yields. Cells appeared to dynamically balance the “push” due to photon flux pressure and reductant generation, with consumption in overall metabolism (“pull” due to demand). However, at 30 °C, all fluorescence parameters declined precipitously, but differential responses were observed depending on N form. Cells enriched with urea at 30 °C showed a smaller decline in fluorescence parameters than cells treated with NO₃⁻ or NH₄⁺, implying that urea might induce a photoprotective mechanism by increasing metabolic “pull”. Full article

The consumption of shellfish contaminated with brevetoxins, a family of ladder-frame polyether toxins formed during blooms of the marine dinoflagellate Karenia brevis, can cause neurotoxic poisoning, leading to gastroenteritis and neurotoxic effects. To rapidly monitor brevetoxin levels in oysters, we generated a broad-spectrum antibody against brevetoxin 2 (PbTx-2), 1 (PbTx-1), and 3 (PbTx-3) and developed a rapid indirect competitive enzyme-linked immunosorbent assay (icELISA). PbTx-2 was reacted with carboxymethoxylamine hemihydrochloride (CMO) to generate a PbTx-2-CMO hapten and reacted with succinic anhydride (HS) to generate the PbTx-2-HS hapten. These haptens were conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen reagents, respectively, using the active ester method. After immunization and cell fusion, a broad-spectrum monoclonal antibody (mAb) termed mAb 1D3 was prepared. The 50% inhibitory concentration (IC₅₀) values of the icELISA for PbTx-2, PbTx-1, and PbTx-3 were 60.71, 52.61, and 51.83 μg/kg, respectively. Based on the broad-spectrum mAb 1D3, an icELISA was developed to determine brevetoxin levels. Using this approach, the limit of detection (LOD) for brevetoxin was 124.22 μg/kg and recoveries ranged between 89.08% and 115.00%, with a coefficient of variation below 4.25% in oyster samples. These results suggest that our icELISA is a useful tool for the rapid monitoring of brevetoxins in oyster samples. Full article

In the last few decades, harmful algal blooms (HABs, also known as “red tides”) have become one of the most detrimental natural phenomena in Florida’s coastal areas. Karenia brevis produces toxins that have harmful effects on humans, fisheries, and ecosystems. In this study, we developed and compared the efficiency of state-of-the-art machine learning models (e.g., XGBoost, Random Forest, and Support Vector Machine) in predicting the occurrence of HABs. In the proposed models the K. brevis abundance is used as the target, and 10 level-02 ocean color products extracted from daily archival MODIS satellite data are used as controlling factors. The adopted approach addresses two main shortcomings of earlier models: (1) the paucity of satellite data due to cloudy scenes and (2) the lag time between the period at which a variable reaches its highest correlation with the target and the time the bloom occurs. Eleven spatio-temporal models were generated, each from 3 consecutive day satellite datasets, with a forecasting span from 1 to 11 days. The 3-day models addressed the potential variations in lag time for some of the temporal variables. One or more of the generated 11 models could be used to predict HAB occurrences depending on availability of the cloud-free consecutive days. Findings indicate that XGBoost outperformed the other methods, and the forecasting models of 5–9 days achieved the best results. The most reliable model can forecast eight days ahead of time with balanced overall accuracy, Kappa coefficient, F-Score, and AUC of 96%, 0.93, 0.97, and 0.98 respectively. The euphotic depth, sea surface temperature, and chlorophyll-a are always among the most significant controlling factors. The proposed models could potentially be used to develop an “early warning system” for HABs in southwest Florida. Full article