Exposure to Aerosolized Algal Toxins in South Florida Increases Short- and Long-Term Health Risk in Drosophila Model of Aging.

Harmful algal blooms (HABs) are a rising health and environmental concern in the United States, particularly in South Florida. Skin contact and the ingestion of contaminated water or fish and other seafood have been proven to have severe toxicity to humans in some cases. However, the impact of aerosolized HAB toxins is poorly understood. In particular, knowledge regarding either the immediate or long-term effects of exposure to aerosolized cyanotoxins produced by freshwater blue-green algae does not exist. The aim of this study was to probe the toxicity of aerosolized cyanobacterial blooms using Drosophila melanogaster as an animal model. The exposure of aerosolized HABs at an early age leads to the most severe long-term impact on health and longevity among all age groups. Young groups and old males showed a strong acute response to HAB exposure. In addition, brain morphological analysis using fluorescence imaging reveals significant indications of brain degeneration in females exposed to aerosolized HABs in early or late stages. These results indicate that one-time exposure to aerosolized HAB particles causes a significant health risk, both immediately and in the long-term. Interestingly, age at the time of exposure plays an important role in the specific nature of the impact of aerosol HABs. As BMAA and microcystin have been found to be the significant toxins in cyanobacteria, the concentration of both toxins in the water and aerosols was examined. BMAA and microcystin are consistently detected in HAB waters, although their concentrations do not always correlate with the severity of the health impact, suggesting the potential contribution from additional toxins present in the aerosolized HAB. This study demonstrates, for the first time, the health risk of exposure to aerosolized HAB, and further highlights the critical need and importance of understanding the toxicity of aerosolized cyanobacteria HAB particles and determining the immediate and long-term health impacts of HAB exposure.

Cyanobacterial neurotoxin BMAA and brain pathology in stranded dolphins.

Dolphin stranding events occur frequently in Florida and Massachusetts. Dolphins are an excellent sentinel species for toxin exposures in the marine environment. In this report we examine whether cyanobacterial neurotoxin, ß-methylamino-L-alanine (BMAA), is present in stranded dolphins. BMAA has been shown to bioaccumulate in the marine food web, including in the muscles and fins of sharks. Dietary exposure to BMAA is associated with the occurrence of neurofibrillary tangles and ß-amyloid plaques in nonhuman primates. The findings of protein-bound BMAA in brain tissues from patients with Alzheimer's disease has advanced the hypothesis that BMAA may be linked to dementia. Since dolphins are apex predators and consume prey containing high amounts of BMAA, we examined necropsy specimens to determine if dietary and environmental exposures may result in the accumulation of BMAA in the brains of dolphins. To test this hypothesis, we measured BMAA in a series of brains collected from dolphins stranded in Florida and Massachusetts using two orthogonal analytical methods: 1) high performance liquid chromatography, and 2) ultra-performance liquid chromatography with tandem mass spectrometry. We detected high levels of BMAA (20-748 µg/g) in the brains of 13 of 14 dolphins. To correlate neuropathological changes with toxin exposure, gross and microscopic examinations were performed on cortical brain regions responsible for acoustico-motor navigation. We observed increased numbers of ß-amyloid+ plaques and dystrophic neurites in the auditory cortex compared to the visual cortex and brainstem. The presence of BMAA and neuropathological changes in the stranded dolphin brain may help to further our understanding of cyanotoxin exposure and its potential impact on human health.

KARENIA: The biology and ecology of a toxic genus.

Karenia is a genus containing at least 12 species of marine unarmored dinoflagellates. Species of the genus can be found throughout the world in both oceanic and coastal waters. They are usually sparse in abundance, but occasionally form large blooms in coastal waters. Most Karenia species produce a variety of toxins that can kill fish and other marine organisms when they bloom. In addition to toxicity, some Karenia blooms cause animal mortalities through the generation of anoxia. At least one species, K. brevis, produces brevetoxin that not only kills fish, marine mammals, and other animals, but also causes Neurotoxic Shellfish Poisoning and respiratory distress in humans. The lipid soluble brevetoxin can biomagnify up the food chain through fish to top carnivores like dolphins, killing them. Karenia dinoflagellates are slow growers, so physical concentrating mechanisms are probably important for the development of blooms. The blooms are highly sporadic in both time and space, although most tend to occur in summer or fall months in frontal regions. At the present time, our understanding of the causes of the blooms and ability to predict them is poor. Given the recent discovery of new species, it is likely that new Karenia species and toxins will be discovered in the future.

Relative risk assessment of cruise ships biosolids disposal alternatives.

A relative risk assessment of biosolids disposal alternatives for cruise ships is presented in this paper. The area of study encompasses islands and marine waters of the Caribbean Sea. The objective was to evaluate relative human health and ecological risks of (a) dewatering/incineration, (b) landing the solids for disposal, considering that in some countries land-disposed solids might be discharged in the near-shore environment untreated, and (c) deep ocean disposal. Input to the Bayesian assessment consisted of professional judgment based on available literature and modeling information, data on constituent concentrations in cruise ship biosolids, and simulations of constituent concentrations in Caribbean waters assuming ocean disposal. Results indicate that human health and ecological risks associated with land disposal and shallow ocean disposal are higher than those of the deep ocean disposal and incineration. For incineration, predicted ecological impacts were lower relative to deep ocean disposal before considering potential impacts of carbon emissions.

Cyanobacterial Blooms and the Occurrence of the neurotoxin beta-N-methylamino-L-alanine (BMAA) in South Florida Aquatic Food Webs.

Recent studies demonstrate that most cyanobacteria produce the neurotoxin beta-N-methylamino-L-alanine (BMAA) and that it can biomagnify in at least one terrestrial food chain. BMAA has been implicated as a significant environmental risk in the development of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Amyotrophic Lateral Sclerosis (ALS). We examined several blooms of cyanobacteria in South Florida, and the BMAA content of resident animals, including species used as human food. A wide range of BMAA concentrations were found, ranging from below assay detection limits to approximately 7000 µg/g, a concentration associated with a potential long-term human health hazard.

Morphological and physiological characteristics of Gephyrocapsa oceanica var. typica Kamptner 1943 in culture experiments: evidence for genotypic variability.

In order to test whether morphological variations within Gephyrocapsa oceanica var. typica Kamptner 1943 reflect genotypic variation or phenotypic responses to environmental conditions, culture experiments of six strains of G. oceanica collected at different locations in the North Atlantic and Mediterranean have been carried out under different temperature and nutrients conditions. All morphological, and physiological data suggest the presence of two species or subspecies within G. oceanica var typica that correspond morphologically to Gephyrocapsa "Larger" and possibly to Gephyrocapsa "Equatorial" as previously defined from Holocene sediments. Given the importance of Gephyrocapsa species for the carbon cycle in the past, genetic studies on this group are of major interest to the understanding of past climate change and plankton evolution.

Human exposure to cyanobacteria and BMAA.

Cyanobacteria are found worldwide, primarily in aquatic habitats. They are increasing in abundance as a result of increasing nutrient inputs from various human activities. Recent data indicate that most cyanobacteria produce the neurotoxin beta-N-methylamino-L-alanine (BMAA), and this toxin can biomagnify UP some food chains to rather high concentrations in animals used as food by humans. BMAA may pose an increasing human health risk.

Viable cell sorting of dinoflagellates by multiparametric flow cytometry.

Electronic cell sorting for isolation and culture of dinoflagellates and other marine eukaryotic phytoplankton was compared to the traditional method of manually picking cells using a micropipette. Trauma to electronically sorted cells was not a limiting factor, as fragile dinoflagellates, such as Karenia brevis (Dinophyceae), survived electronic cell sorting to yield viable cells. The rate of successful isolation of large-scale (> 4 litres) cultures was higher for manual picking than for electronic cell sorting (2% vs 0.5%, respectively). However, manual picking of cells is more labor intensive and time consuming. Most manually isolated cells required repicking, as the cultures were determined not to be unialgal after a single round of isolation; whereas, no cultures obtained in this study from electronic single-cell sorting required resorting. A broad flow cytometric gating logic was employed to enhance species diversity. The percentages of unique genotypes produced by manual picking or electronic cell sorting were similar (57% vs 54%, respectively), and each approach produced a variety of dinoflagellate or raphidophyte genera. Alternatively, a highly restrictive gating logic was successfully used to target K. brevis from a natural bloom sample. Direct electronic single-cell sorting was more successful than utilizing a pre-enrichment sort followed by electronic single-cell sorting. The appropriate recovery medium may enhance the rate of successful isolations. Seventy percent of isolated cells were recovered in a new medium (RE) reported here, which was optimized for axenic dinoflagellate cultures. The greatest limiting factor to the throughput of electronic cell sorting is the need for manual postsort culture maintenance and assessment of the large number of isolated cells. However, when combined with newly developed automated methods for growth screening, electronic single-cell sorting has the potential to accelerate the discovery of new algal strains.

Centers for Oceans and Human Health: a unified approach to the challenge of harmful algal blooms.

BACKGROUND: Harmful algal blooms (HABs) are one focus of the national research initiatives on Oceans and Human Health (OHH) at NIEHS, NOAA and NSF. All of the OHH Centers, from the east coast to Hawaii, include one or more research projects devoted to studying HAB problems and their relationship to human health. The research shares common goals for understanding, monitoring and predicting HAB events to protect and improve human health: understanding the basic biology of the organisms; identifying how chemistry, hydrography and genetic diversity influence blooms; developing analytical methods and sensors for cells and toxins; understanding health effects of toxin exposure; and developing conceptual, empirical and numerical models of bloom dynamics. RESULTS: In the past several years, there has been significant progress toward all of the common goals. Several studies have elucidated the effects of environmental conditions and genetic heterogeneity on bloom dynamics. New methods have been developed or implemented for the detection of HAB cells and toxins, including genetic assays for Pseudo-nitzschia and Microcystis, and a biosensor for domoic acid. There have been advances in predictive models of blooms, most notably for the toxic dinoflagellates Alexandrium and Karenia. Other work is focused on the future, studying the ways in which climate change may affect HAB incidence, and assessing the threat from emerging HABs and toxins, such as the cyanobacterial neurotoxin beta-N-methylamino-L-alanine. CONCLUSION: Along the way, many challenges have been encountered that are common to the OHH Centers and also echo those of the wider HAB community. Long-term field data and basic biological information are needed to develop accurate models. Sensor development is hindered by the lack of simple and rapid assays for algal cells and especially toxins. It is also critical to adequately understand the human health effects of HAB toxins. Currently, we understand best the effects of acute toxicity, but almost nothing is known about the effects of chronic, subacute toxin exposure. The OHH initiatives have brought scientists together to work collectively on HAB issues, within and across regions. The successes that have been achieved highlight the value of collaboration and cooperation across disciplines, if we are to continue to advance our understanding of HABs and their relationship to human health.

Determination of brevetoxin in recent marine sediments.

Harmful algal blooms (HAB) of Karenia brevis (K. brevis) produce a suite of lipid soluble polyether brevetoxins, known to cause environmental, health and economic ill effects. There is evidence that K. brevis has increased in abundance over the past 50 years, but the dataset is incomplete. The objective of this paper was to analyze sediment from an area where K. brevis blooms have occurred and investigate if these compounds are incorporated into the underlying sediment, thus potentially allowing the use of brevetoxins as an indicator of past K. Brevis blooms. The results from LC-ESI-MS-MS analyses of brevetoxin analogs detected in surficial sediments from three sites (Fort Meyers Beach [FMB], Big Hickory Pass [BHP] and Big Carlos Pass [BCP]) along the Southwest Florida coastline with prior HAB history are promising. The analogs detected from BHP sediments were PbTx-2 and PbTx-3 with values of 0.81 and 3.1 ng g(-1) dry sediment, respectively. The detected PbTx-2 from BCP was 3.6 ng g(-1) dry sediment, while the detected PbTx-3 from BCP was 9.7 ng g(-1) dry sediment. PbTx-3 was only detected at the FMB site (2.7 ng g(-1) dry sediment). The detection of brevetoxins in recent sediments where K. brevis have occurred indicates brevetoxin incorporation into marine sediments.

Ocean urea fertilization for carbon credits poses high ecological risks.

The proposed plan for enrichment of the Sulu Sea, Philippines, a region of rich marine biodiversity, with thousands of tonnes of urea in order to stimulate algal blooms and sequester carbon is flawed for multiple reasons. Urea is preferentially used as a nitrogen source by some cyanobacteria and dinoflagellates, many of which are neutrally or positively buoyant. Biological pumps to the deep sea are classically leaky, and the inefficient burial of new biomass makes the estimation of a net loss of carbon from the atmosphere questionable at best. The potential for growth of toxic dinoflagellates is also high, as many grow well on urea and some even increase their toxicity when grown on urea. Many toxic dinoflagellates form cysts which can settle to the sediment and germinate in subsequent years, forming new blooms even without further fertilization. If large-scale blooms do occur, it is likely that they will contribute to hypoxia in the bottom waters upon decomposition. Lastly, urea production requires fossil fuel usage, further limiting the potential for net carbon sequestration. The environmental and economic impacts are potentially great and need to be rigorously assessed.

Long-term increase in Karenia brevis abundance along the Southwest Florida Coast.

Data collected along the southwest coast of Florida between Tampa Bay and Sanibel Island on the abundance of the toxic dinoflagellate Karenia brevis from 1954 to 2002 were examined for spatial and temporal patterns. K. brevis was found to be approximately 20-fold more abundant within 5 km of the shoreline than 20-30 km offshore. Overall, K. brevis was approximately 13-18-fold more abundant in 1994-2002 than in 1954-1963. In 1954-1963, K. brevis occurred primarily in the fall months. In 1994-2002, it was more abundant not only in the fall, but also in the winter and spring months. It is hypothesized that greater nutrient availability in the ecosystem is the most likely cause of this increase in K. brevis biomass, and the large increase in the human population and its activities in South Florida over the past half century is a major factor.