Karenia Brevis Research Articles

Nitrogen-enriched discharges from a highly managed watershed intensify red tide (Karenia brevis) blooms in southwest Florida

Karenia brevis blooms on Florida's Gulf Coast severely affect regional ecosystems, coastal economies, and public health, and formulating effective management and policy strategies to address these blooms requires an advanced understanding of the processes driving them. Recent research suggests that natural processes explain offshore bloom initiation and shoreward transport, while anthropogenic nutrient inputs may intensify blooms upon arrival along the coast. However, past correlation studies have failed to detect compelling evidence linking coastal blooms to watershed covariates indicative of anthropogenic inputs. We explain why correlation is neither necessary nor sufficient to demonstrate a causal relationship—i.e., a persistent pattern of interaction governed by deterministic rules—and pursue an empirical investigation leveraging the fact that systematic temporal patterns may reveal systematic cause-and-effect relationships. Using time series derived from in-situ sample data, we applied singular spectrum analysis—a non-parametric spectral decomposition method—to recover deterministic signals in the dynamics of K. brevis blooms and upstream water quality and discharge covariates in the Charlotte Harbor region between 2012 and 2021. Next, we applied causal analysis methods based on chaos theory—i.e., convergent cross-mapping and S-mapping—to detect and quantify persistent, state-dependent interaction regimes between coastal blooms and watershed covariates. We discovered that nitrogen-enriched Caloosahatchee River discharges have consistently intensified K. brevis blooms to varying degrees over time. River discharge was typically most influential at the earliest stages of blooms, while total nitrogen concentrations exerted the strongest influence during blooms' growth/maintenance stages. These results indicate that discharges and nitrogen inputs influence blooms through distinct yet synergistic causal mechanisms. Additionally, we traced this anthropogenic influence upstream to Lake Okeechobee (which discharges to the Caloosahatchee River) and the Kissimmee River basin (which drains into Lake Okeechobee), suggesting that watershed-scale nutrient management and modifications to Lake Okeechobee discharge protocols will likely be necessary to mitigate coastal blooms.

Prescreening-Based Subset Selection for Improving Predictions of Earth System Models With Application to Regional Prediction of Red Tide.

We present the ensemble method of prescreening-based subset selection to improve ensemble predictions of Earth system models (ESMs). In the prescreening step, the independent ensemble members are categorized based on their ability to reproduce physically-interpretable features of interest that are regional and problem-specific. The ensemble size is then updated by selecting the subsets that improve the performance of the ensemble prediction using decision relevant metrics. We apply the method to improve the prediction of red tide along the West Florida Shelf in the Gulf of Mexico, which affects coastal water quality and has substantial environmental and socioeconomic impacts on the State of Florida. Red tide is a common name for harmful algal blooms that occur worldwide, which result from large concentrations of aquatic microorganisms, such as dinoflagellate Karenia brevis, a toxic single celled protist. We present ensemble method for improving red tide prediction using the high resolution ESMs of the Coupled Model Intercomparison Project Phase 6 (CMIP6) and reanalysis data. The study results highlight the importance of prescreening-based subset selection with decision relevant metrics in identifying non-representative models, understanding their impact on ensemble prediction, and improving the ensemble prediction. These findings are pertinent to other regional environmental management applications and climate services. Additionally, our analysis follows the FAIR Guiding Principles for scientific data management and stewardship such that data and analysis tools are findable, accessible, interoperable, and reusable. As such, the interactive Colab notebooks developed for data analysis are annotated in the paper. This allows for efficient and transparent testing of the results' sensitivity to different modeling assumptions. Moreover, this research serves as a starting point to build upon for red tide management, using the publicly available CMIP, Coordinated Regional Downscaling Experiment (CORDEX), and reanalysis data.

Characterization of Atmospheric Processes of Brevetoxins in Sea Spray Aerosols from Red Tide Events.

Atmospheric processes can affect the longevity of harmful toxins in sea spray aerosols (SSA). This study characterized the degradation of brevetoxin (BTx) in SSA under different environmental conditions. The samples of seawater collected during a Karenia brevis bloom in Manasota, Florida, were nebulized into a large outdoor photochemical chamber to mimic the atmospheric oxidation of aerosolized toxins and then aged in the presence or absence of sunlight and/or O3. Aerosol samples were collected during the aging process using a Particle-Into-Liquid Sampler. Their BTx concentrations were measured using an enzyme-linked immuno-sorbent assay (ELISA) and high-performance liquid chromatography/tandem mass spectroscopy. The BTx ozonolysis rate constant measured by ELISA was 5.74 ± 0.21 × 103 M-1 s-1. The corresponding lifetime for decay of 87.5% BTx in the presence of 20 ppb of O3 was 7.08 ± 0.26 h, suggesting that aerosolized BTx can still travel long distances at night before SSA deposition. BTx concentrations in SSA decreased more rapidly in the presence of sunlight than in its absence due to oxidation with photochemically produced OH radicals.

Quantifying Karenia brevis bloom severity and respiratory irritation impact along the shoreline of Southwest Florida.

Nearly all annual blooms of the toxic dinoflagellate Karenia brevis (K. brevis) pose a serious threat to coastal Southwest Florida. These blooms discolor water, kill fish and marine mammals, contaminate shellfish, cause mild to severe respiratory irritation, and discourage tourism and recreational activities, leading to significant health and economic impacts in affected communities. Despite these issues, we still lack standard measures suitable for assessing bloom severity or for evaluating the efficacy of modeling efforts simulating bloom initiation and intensity. In this study, historical cell count observations along the southwest Florida shoreline from 1953 to 2019 were used to develop monthly and annual bloom severity indices (BSI). Similarly, respiratory irritation observations routinely reported in Sarasota and Manatee Counties from 2006 to 2019 were used to construct a respiratory irritation index (RI). Both BSI and RI consider spatial extent and temporal evolution of the bloom, and can be updated routinely and used as objective criteria to aid future socioeconomic and scientific studies of K. brevis. These indices can also be used to help managers and decision makers both evaluate the risks along the coast during events and design systems to better respond to and mitigate bloom impacts. Before 1995, sampling was done largely in response to reports of discolored water, fish kills, or respiratory irritation. During this timeframe, lack of sampling during the fall, when blooms typically occur, generally coincided with periods of more frequent-than-usual offshore winds. Consequently, some blooms may have been undetected or under-sampled. As a result, the BSIs before 1995 were likely underestimated and cannot be viewed as accurately as those after 1995. Anomalies in the frequency of onshore wind can also largely account for the discrepancies between BSI and RI during the period from 2006 to 2019. These findings highlighted the importance of onshore wind anomalies when predicting respiratory irritation impacts along beaches.

Nitrogen-Enriched Discharges from a Highly Managed Watershed Intensify Red Tide (Karenia Brevis) Blooms in Southwest Florida

Nitrogen-Enriched Discharges from a Highly Managed Watershed Intensify Red Tide (Karenia Brevis) Blooms in Southwest Florida

Effects of the toxic dinoflagellate Karenia brevis on the development of the marine gastropod Crepidula fornicata

With harmful algal blooms increasing in frequency and range, it is important to study how exposure to harmful algae may affect the development of marine organisms. Our studies examined the susceptibility of the early life stages of the Atlantic slipper limpet Crepidula fornicata to the toxic red tide dinoflagellate Karenia brevis (strain FWC1010), testing the effects of exposure to 3 algal bloom concentrations (100, 1000, or 5000 cells ml-1), representing medium to high bloom conditions in nature, on survival, growth, clearance rates, metamorphosis, and post-metamorphic survival and growth. Larvae were given either a mixed diet of K. brevis together with the non-toxic phytoplankton Tisochrysis lutea (clone TISO), a unialgal diet of K. brevis, or a control diet of unialgal TISO. In some experiments, larvae were raised to metamorphosis on the different diets and were then observed as juveniles in a non-toxic environment to see if there were any latent effects of larval exposure to K. brevis on subsequent juvenile survival or growth (in terms of both shell length and percent inorganic weight). There were no significant effects of larval exposure to K. brevis on larval or juvenile survival. However, we observed several notable sub-lethal effects of larval exposure to high K. brevis concentrations (1000 and 5000 cells ml-1), including slower larval shell growth rates, delayed competence for metamorphosis, lower responses to natural metamorphic cues, and reduced rates of post-metamorphic growth (in terms of both shell length per day and percent inorganic weight) when juveniles were reared under control conditions.

Dynamics of cell death across growth stages and the diel cycle in the dinoflagellate Karenia brevis.

Recent evidence suggests that programmed cell death (PCD) can play a role in stress-induced decline and termination of harmful algal blooms. However, components of the PCD cascade, i.e. reactive oxygen species (ROS) and caspase-like activity, have also been observed in the absence of exogenous stress, where their activities and functions remain unclear. Here, we characterized the variability of prevalence of cell death, ROS, and caspase-like activity at different growth phases and diel cycles in cultures of dinoflagellate Karenia brevis. Results show that ROS percentages increased with culture age and fluctuated in a phasing diel pattern, while caspase-like activity was observed throughout growth. In actively growing K.brevis cells, PCD components may be involved in key metabolic processes, while in stationary phase they may relate to stress acclimation. The circadian diel pattern of ROS may be explained by the balance between the metabolic generation of ROS and circadian rhythmicity of antioxidant enzymes. Overall, this work highlights not only the involvement of PCD components in the growth of marine phytoplankton, but the importance of understanding mechanisms controlling their accumulation, which would help to better interpret their presence in the field.

Using localized Twitter activity to assess harmful algal bloom impacts of Karenia brevis in Florida, USA

Red tide blooms of the dinoflagellate Karenia brevis (K. brevis) produce toxic coastal conditions that can impact marine organisms and human health, while also affecting local economies. During the extreme Florida red tide event of 2017–2019, residents and visitors turned to social media platforms to both receive disaster-related information and communicate their own sentiments and experiences. This was the first major red tide event since the ubiquitous use of social media, thus providing unique crowd-sourced reporting of red tide impacts. We evaluated the spatial and temporal accuracy of red tide topic activity on Twitter, taking tweet sentiments and user types (e.g. media, citizens) into consideration, and compared tweet activity with reported red tide conditions, such as K. brevis cell counts, levels of dead fish and respiratory irritation on local beaches. The analysis was done on multiple levels with respect to both locality (e.g., entire Gulf coast, county-level, city-level, zip code tabulation areas) and temporal frequencies (e.g. daily, every three days, weekly), resulting in strong correlations between local per-capita Twitter activity and the actual red tide conditions observed in the area. Moreover, an association was observed between proximity to the affected coastal areas and per-capita counts for relevant tweets. Results show that Twitter presents a trustworthy reflection of the red tide’s local impacts and development over time, and can potentially augment the already existing tools for efficient assessment and a more coordinated response to the disaster.

Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay.

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.

Guidance Level for Brevetoxins in French Shellfish

Brevetoxins (BTXs) are marine biotoxins responsible for neurotoxic shellfish poisoning (NSP) after ingestion of contaminated shellfish. NSP is characterized by neurological, gastrointestinal and/or cardiovascular symptoms. The main known producer of BTXs is the dinoflagellate Karenia brevis, but other microalgae are also suspected to synthesize BTX-like compounds. BTXs are currently not regulated in France and in Europe. In November 2018, they have been detected for the first time in France in mussels from a lagoon in the Corsica Island (Mediterranean Sea), as part of the network for monitoring the emergence of marine biotoxins in shellfish. To prevent health risks associated with the consumption of shellfish contaminated with BTXs in France, a working group was set up by the French Agency for Food, Environmental and Occupational Health & Safety (Anses). One of the aims of this working group was to propose a guidance level for the presence of BTXs in shellfish. Toxicological data were too limited to derive an acute oral reference dose (ARfD). Based on human case reports, we identified two lowest-observed-adverse-effect levels (LOAELs). A guidance level of 180 µg BTX-3 eq./kg shellfish meat is proposed, considering a protective default portion size of 400 g shellfish meat.

Young-Of-The-Year Stone Crab (Genus Menippe) Recruitment in the Gulf of Mexico Off Florida: Key Shallow-Water Hotspots

Knowledge of juvenile recruitment (defined here as the quantitative addition of early benthic life stages to a local population) is important for conserving heavily harvested species and their critical habitats. Stone crabs (genus Menippe) are commercially and ecologically important throughout the Gulf of Mexico nearshore waters off Florida, but very little is known about their recruitment. Using standard commercial stone crab traps, megalopal and postsettlement juvenile (collectively, young-of-the-year, “YOY”) stone crabs were sampled at multiple spatial scales and for multiple stone crab generations to characterize geographical, seasonal, and interannual variation in their distribution and relative abundance (numbers collected on traps) in the Gulf of Mexico off peninsular Florida and north of the Florida Keys (the “Florida Gulf”). The influences of potentially relevant oceanographic and biological variables [temperature, salinity, benthic community on the traps (trap fouling community), depth, distance from shore] on YOY stone crabs were investigated at locations distributed throughout the study area. Trap fouling communities had never been analyzed in detail prior to this study; an importance index was developed to quantify the seasonality, commonality, and density of the fouling communities and organisms composing the communities. Continuous, long-term data from the Tampa Bay location allowed investigation of the effects of biological relationships [relative abundance of female Menippe carrying eggs (ovigerous), occurrences of red tide (Karenia brevis, a toxic dinoflagellate) blooms] and meteorological events [tropical cyclones, El Niño Southern Oscillation (ENSO) occurrences] on temporal patterns of variation in recruitment. High relative abundances of YOY stone crabs were collected off peninsular Florida from dense, complex benthic biota that grew on crab traps located off large, pristine estuaries in relatively turbid water less than 5 m deep, where salinity ranged 24–36 and water temperature averaged 29°C–32°C. Two major recruitment locations, stable through decades, consistently accounted for approximately 50% of the YOY stone crabs collected; two secondary recruitment locations similarly accounted for another 25%. Relative abundance of YOY stone crabs was highest nearshore at the major recruitment locations, particularly during years of high relative abundance. Approximately 65%–75% of the recruits were collected August to October; however, the timing of both peak relative abundance and lowest relative abundance shifted to later in the year as latitude decreased. Patterns of change among months in relative abundance differed among locations within years and among years within locations. Water temperature, salinity, and trap fouling community were the important determinants of temporal and spatial variation in YOY stone crab relative abundance; depth and distance from shore were also important in areas where they varied notably among stations within locations and across broader expanses of the study area. At Tampa Bay, a sharp increase in relative abundance of ovigerous females in spring was followed by a similarly sharp increase in YOY stone crab relative abundance (principally stage 3–5 crabs) 3 mo later; whereas a sharp decrease in ovigerous female relative abundance in autumn was followed by a similarly sharp decrease in YOY stone crab relative abundance 1 mo later. Coincidence of the normal autumnal decrease in YOY stone crab relative abundance with red tides and tropical cyclones prevented assigning clear relationships between seasonal change in relative abundance and these potential external influences. Annual decreases in relative abundance were significantly related to the occurrence of tropical cyclones that came near Tampa Bay during the previous year. Twice, confluences of multiple hurricanes, timely red tides, and ENSO events were followed by nearly complete, 1- to 2-y recruitment collapses. In contrast, a single meteorologically and oceanographically highly anomalous year coincided with exceptionally high abundances of both YOY and ovigerous female stone crabs. A relationship between YOY stone crabs and subsequent fishery harvest was not evident, possibly due to a variety of fishery practices. Because stone crabs are subjected to intense fishing pressure throughout the Florida Gulf, the YOY stone crab recruitment grounds and their associated estuaries should be protected from the effects of development, agriculture, aquaculture, and commercial and recreational crabbing.

Machine Learning Classification Algorithms for Predicting Karenia brevis Blooms on the West Florida Shelf

Harmful algal blooms (HABs), events that kill fish, impact human health in multiple ways, and contaminate water supplies, have increased in frequency, magnitude, and impacts in numerous marine and freshwaters around the world. Blooms of the toxic dinoflagellate Karenia brevis have resulted in thousands of tons of dead fish, deaths to many other marine organisms, numerous respiratory-related hospitalizations, and tens to hundreds of millions of dollars in economic damage along the West Florida coast in recent years. Four types of machine learning algorithms, Support Vector Machine (SVM), Relevance Vector Machine (RVM), Naïve Bayes classifier (NB), and Artificial Neural Network (ANN), were developed and compared in their ability to predict these blooms. Comparing the 21 year monitoring dataset of K. brevis abundance, RVM and NB were found to have better skills in bloom prediction than the other two approaches. The importance of upwelling-favorable northerly winds in increasing K. brevis probability, and of onshore westerly winds in preventing blooms from dispersing offshore, were quantified using RVM, and all models were used to explore the importance of large river flows and the nutrients they supply in regulating blooms. These models provide new tools for management of these devastating algal blooms.

Effects of Multiple Karenia brevis Red Tide Blooms on a Common Bottlenose Dolphin (Tursiops truncatus) Prey Fish Assemblage: Patterns of Resistance and Resilience in Sarasota Bay, Florida

Red tide blooms caused by the toxic dinoflagellate Karenia brevis are natural disturbance events that occur regularly along Florida’s west coast, often resulting in massive fish kills and marine mammal, seabird, and sea turtle mortalities. Limited prior work on the ecological effects of red tides suggests they play an important role in structuring ecosystem dynamics and regulating communities, however specific effects on prey populations and potential alterations to predator-prey interactions are unknown. We surveyed the prey fish assemblage of a top marine predator, the common bottlenose dolphin (Tursiops truncatus), in shallow seagrass habitat in Sarasota Bay, Florida, during 2004–2019, collecting data on prey density, species composition, K. brevis cell densities, and environmental variables. Across eight distinct red tide bloom events, resistance, resilience, and the ecological effects on the prey assemblage varied depending on bloom intensity, season, and frequency. Prey assemblage structure showed significant and distinct short-term shifts during blooms independent of the normal seasonal shifts in prey structure seen during non-bloom conditions. Canonical correspondence analysis indicated a strong influence of K. brevis density on assemblage structure. Blooms occurring primarily in the summer were associated with less initial prey resistance and higher than average annual catch per unit effort (CPUE) 1–3 years following bloom cessation, with bloom frequency prolonging the time needed to reach higher than average annual CPUE. Regardless of season, recovery to pre-bloom prey abundances occurred within 1 year. Sample-based rarefaction and extrapolation indicated significant differences in prey diversity among summer bloom events. This study is a first step in identifying differences in resistance, resilience, and the ecological effects of multiple red tide bloom events of various temporal scales and intensity on a dolphin prey assemblage. Improved understanding of the influence of red tides on estuarine structural dynamics and function can better inform management, and potentially guide mitigation efforts post-bloom.

A systematic review of waterborne and water-related disease in animal populations of Florida from 1999-2019.

Florida's waters are a reservoir for a host of pathogens and toxins. Many of these microorganisms cause water-related diseases in people that are reportable to the Florida Department of Health. Our objective in this review was to ascertain which water-related pathogens and toxins of public health importance have been found in animal populations in Florida over the last twenty years. Nineteen databases were searched, including PubMed and Web of Science Core Collection, using keywords and search terms for the waterborne diseases, water-related vector-borne diseases, and water-based toxins reportable to the Florida Department of Health. For inclusion, peer-reviewed journal articles were to be written in English, published between January 1, 1999 and December 31, 2019, and contain primary research findings documenting at least one of the water-related pathogens or toxins of interest in an animal population within Florida during this same time frame. Of over eight thousand initial search results, 65 studies were included for final analysis. The most common animal types implicated in the diseases of interest included marine mammals, fish and shellfish, wild birds, and livestock. Toxins or pathogens most often associated with these animals included toxin-producer Karenia brevis, vibriosis, Escherichia coli, and Salmonellosis. Findings from this review elucidate the water-related disease-causing pathogens and toxins which have been reported within animal populations in recent Florida history. As most of these diseases are zoonotic, our results suggest a One Health approach is necessary to support and maintain healthy water systems throughout the state of Florida for the protection of both human and animal populations.

Florida’s Harmful Algal Bloom (HAB) Problem: Escalating Risks to Human, Environmental and Economic Health With Climate Change

Harmful Algal Blooms (HABs) pose unique risks to the citizens, stakeholders, visitors, environment and economy of the state of Florida. Florida has been historically subjected to reoccurring blooms of the toxic marine dinoflagellate Karenia brevis (C. C. Davis) G. Hansen & Moestrup since at least first contact with explorers in the 1500’s. However, ongoing immigration of more than 100,000 people year–1 into the state, elevated population densities in coastal areas with attendant rapid, often unregulated development, coastal eutrophication, and climate change impacts (e.g., increasing hurricane severity, increases in water temperature, ocean acidification and sea level rise) has likely increased the occurrence of other HABs, both freshwater and marine, within the state as well as the number of people impacted by these blooms. Currently, over 75 freshwater, estuarine, coastal and marine HAB species are routinely monitored by state agencies. While only blooms of K. brevis, the dinoflagellate Pyrodinium bahamense (Böhm) Steidinger, Tester, and Taylor and the diatom Pseudo-nitzschia spp. have resulted in closure of commercial shellfish beds, other HAB species, including freshwater and marine cyanobacteria, pose either imminent or unknown risks to human, environmental and economic health. HAB related human health risks can be classified into those related to consumption of contaminated shellfish and finfish, consumption of or contact with bloom or toxin contaminated water or exposure to aerosolized HAB toxins. While acute human illnesses resulting from consumption of brevetoxin-, saxitoxin-, and domoic acid-contaminated commercial shellfish have been minimized by effective monitoring and regulation, illnesses due to unregulated toxin exposures, e.g., ciguatoxins and cyanotoxins, are not well documented or understood. Aerosolized HAB toxins potentially impact the largest number of people within Florida. While short-term (days to weeks) impacts of aerosolized brevetoxin exposure are well documented (e.g., decreased respiratory function for at-risk subgroups such as asthmatics), little is known of longer term (>1 month) impacts of exposure or the risks posed by aerosolized cyanotoxin [e.g., microcystin, β-N-methylamino-L-alanine (BMAA)] exposure. Environmental risks of K. brevis blooms are the best studied of Florida HABs and include acute exposure impacts such as significant dies-offs of fish, marine mammals, seabirds and turtles, as well as negative impacts on larval and juvenile stages of many biota. When K. brevis blooms are present, brevetoxins can be found throughout the water column and are widespread in both pelagic and benthic biota. The presence of brevetoxins in living tissue of both fish and marine mammals suggests that food web transfer of these toxins is occurring, resulting in toxin transport beyond the spatial and temporal range of the bloom such that impacts of these toxins may occur in areas not regularly subjected to blooms. Climate change impacts, including temperature effects on cell metabolism, shifting ocean circulation patterns and changes in HAB species range and bloom duration, may exacerbate these dynamics. Secondary HAB related environmental impacts are also possible due to hypoxia and anoxia resulting from elevated bloom biomass and/or the decomposition of HAB related mortalities. Economic risks related to HABs in Florida are diverse and impact multiple stakeholder groups. Direct costs related to human health impacts (e.g., increased hospital visits) as well as recreational and commercial fisheries can be significant, especially with wide-spread sustained HABs. Recreational and tourism-based industries which sustain a significant portion of Florida’s economy are especially vulnerable to both direct (e.g., declines in coastal hotel occupancy rates and restaurant and recreational users) and indirect (e.g., negative publicity impacts, associated job losses) impacts from HABs. While risks related to K. brevis blooms are established, Florida also remains susceptible to future HABs due to large scale freshwater management practices, degrading water quality, potential transport of HABs between freshwater and marine systems and the state’s vulnerability to climate change impacts.

Geospatial distribution of hypoxia associated with a Karenia brevis bloom

In 2018, the presence of bottom water hypoxia along the SW Florida coast was investigated during a bloom of the toxic dinoflagellate Karenia brevis. The bloom was first detected in November 2017. Monitoring of oxygen levels and bloom densities was carried out in 2018 and 2019 using sampling grids. Vertical profiles indicated a pycnocline at 3–4 m where warmer, lower salinity water was at the surface, while the deeper hypoxic layer was colder with higher salinity. There were significantly higher abundances of K. brevis in the surface water compared to the hypoxic bottom water in September 2018. At two fixed sites, dissolved oxygen was measured continuously showing hypoxic conditions during that month. Geospatial analysis of vertical profile data yielded an estimate that the hypoxic layer covered an area of at least 655 km2. The possible influences of red tides on hypoxic conditions along the coast of the eastern Gulf of Mexico are discussed within the context of the 2018 K. brevis bloom event. Hypoxia occurring in parallel to a red tide bloom is more likely to occur with warmer ocean temperatures and increased fluxes of nutrients and fresh water to the Gulf of Mexico after hurricanes.

Utility of Red Tide (Karenia brevis) Monitoring Data as a Predictive Tool to Estimate Brevetoxin Accumulation in Live, Free-Ranging Marine Mammals

Exposure of common bottlenose dolphins (Tursiops truncatus) to brevetoxins (PbTx) produced by blooms of the toxic phytoplanktonKarenia brevisfrequently results in severe health impacts, including illness and large-scale mortality events. Although PbTx accumulation in dead-stranded dolphins is well documented, there are limited data for corresponding brevetoxin exposure in live dolphins. In addition, the severity of impacts on living survivors of such toxic blooms is difficult to assess due to a lack of data on the relationship betweenK. brevisbloom severity and corresponding PbTx concentrations in exposed animals. Here we present results of PbTx analysis of urine, serum, milk, gastric fluid, and feces samples collected from live, free-ranging dolphins (n= 253) from Sarasota Bay, Florida during 2000–2018, and investigate the relationship between PbTx concentrations detected and correspondingK. breviscell abundances that are temporally (within 30 days) and spatially (within 16 km) associated with each individual. We found that 28% of dolphins were associated with elevatedK. brevisabundances (10,000–60,000,000 cells/L), with 41% (n= 104) of dolphins testing positive for PbTx in at least one sample type. The proportion of PbTx-positive animals was significantly greater in animals exposed to elevated cell abundances vs. those exposed to background cell abundances (<10,000 cells/L), with 60 and 34% testing positive, respectively (p< 0.001). PbTx was detected most frequently in feces (57%,n= 38), followed by gastric (35%,n= 37), urine (32%,n= 55), and blood (7%,n= 17). PbTx concentrations by sample type were highest in feces (2–231 ng/g; mean 46), followed by urine (0.8–90 ng/g; mean 7.2), gastric (0.8–61 ng/g; mean 12), and blood (0.3–5 ng/g; mean 1.3). Regression analyses ofK. breviscell abundance as an index of exposure vs. corresponding PbTx concentration found no statistically significant relationship for feces (p= 0.120), gastric (p= 0.349), urine (p= 0.053), or blood (p= 0.729) samples. PbTx concentrations typically ranged over two orders of magnitude between minimum and maximum values and did not scale with corresponding indices of exposure, which ranged over three orders of magnitude or more. Our results indicate thatK. breviscell abundance alone is a poor predictor of brevetoxin accumulation in bottlenose dolphins, and suggest that alternative methods (e.g., endocrine or immunological biomarkers) should be investigated as more appropriate methods for determining the severity of health impacts due to red tides.

Essential components of the xanthophyll cycle differ in high and low toxin Karenia brevis

The dinoflagellate Karenia brevis, blooms annually in the Gulf of Mexico, producing a suite of neurotoxins known as the brevetoxins. The cellular toxin content of K. brevis, however, is highly variable between or even within strains. Herein, we investigate physiological differences between high (KbHT) and low (KbLT) toxin producing cultures both derived from the Wilson strain, related to energy-dependent quenching (qE) by photosystem II, and reduced thiol content of the proteome. We demonstrate that gene and protein expression of the xanthophyll cycle enzyme diadinoxanthin de-epoxidase (Dde) and monogalactosyldiacylglycerol (MGDG) synthase are not significantly different in the two cultures. Using redox proteomics, we report a significantly higher reduced cysteine content in the low toxin proteome, including plastid localized thioredoxin reductase (Trx) which can result in inactivation of Dde and activation of MGDG synthase. We also report significant differences in the lipidomes of KbHT and KbLT with respect to MGDG, which facilitates the xanthophyll cycle.

Investigating an increase in Florida manatee mortalities using a proteomic approach

Two large-scale Florida manatee (Trichechus manatus latirostris) mortality episodes were reported on separate coasts of Florida in 2013. The east coast mortality episode was associated with an unknown etiology in the Indian River Lagoon (IRL). The west coast mortality episode was attributed to a persistent Karenia brevis algal bloom or ‘red tide’ centered in Southwest Florida. Manatees from the IRL also had signs of cold stress. To investigate these two mortality episodes, two proteomic experiments were performed, using two-dimensional difference in gel electrophoresis (2D-DIGE) and isobaric tags for relative and absolute quantification (iTRAQ) LC–MS/MS. Manatees from the IRL displayed increased levels of several proteins in their serum samples compared to controls, including kininogen-1 isoform 1, alpha-1-microglobulin/bikunen precursor, histidine-rich glycoprotein, properdin, and complement C4-A isoform 1. In the red tide group, the following proteins were increased: ceruloplasmin, pyruvate kinase isozymes M1/M2 isoform 3, angiotensinogen, complement C4-A isoform 1, and complement C3. These proteins are associated with acute-phase response, amyloid formation and accumulation, copper and iron homeostasis, the complement cascade pathway, and other important cellular functions. The increased level of complement C4 protein observed in the red tide group was confirmed through the use of Western Blot.

Calcium Increase and Substance P Release Induced by the Neurotoxin Brevetoxin-1 in Sensory Neurons: Involvement of PAR2 Activation through Both Cathepsin S and Canonical Signaling.

Red tides involving Karenia brevis expose humans to brevetoxins (PbTxs). Oral exposition triggers neurotoxic shellfish poisoning, whereas inhalation induces a respiratory syndrome and sensory disturbances. No curative treatment is available and the pathophysiology is not fully elucidated. Protease-activated receptor 2 (PAR2), cathepsin S (Cat-S) and substance P (SP) release are crucial mediators of the sensory effects of ciguatoxins (CTXs) which are PbTx analogs. This work explored the role of PAR2 and Cat-S in PbTx-1-induced sensory effects and deciphered the signaling pathway involved. We performed calcium imaging, PAR2 immunolocalization and SP release experiments in monocultured sensory neurons or co-cultured with keratinocytes treated with PbTx-1 or P-CTX-2. We demonstrated that PbTx-1-induced calcium increase and SP release involved Cat-S, PAR2 and transient receptor potential vanilloid 4 (TRPV4). The PbTx-1-induced signaling pathway included protein kinase A (PKA) and TRPV4, which are compatible with the PAR2 biased signaling induced by Cat-S. Internalization of PAR2 and protein kinase C (PKC), inositol triphosphate receptor and TRPV4 activation evoked by PbTx-1 are compatible with the PAR2 canonical signaling. Our results suggest that PbTx-1-induced sensory disturbances involve the PAR2-TRPV4 pathway. We identified PAR2, Cat-S, PKA, and PKC that are involved in TRPV4 sensitization induced by PbTx-1 in sensory neurons.