Symptom frequency and exposure to a cyanobacteria bloom in Florida.

This investigation was undertaken to characterize health effects associated with a major bloom of blue-green algae due to the proliferation Microcystis aeruginosa that occurred in Florida in 2018. Cyanobacteria produce multiple toxins, including the potent hepatotoxic microcystins (MCs), that have been reported to cause illness in exposed persons worldwide. Widespread exposure to toxins released by blue-green algae during the 2018 bloom was shown by the presence of MCs in the nasal passages of 95 percent of the individuals studied previously in south Florida (Schaefer et al., 2020). The current analyses were conducted to determine whether self-reported symptoms were associated with activity patterns, direct contact with water, residential, recreational, and occupational exposure. The 125 persons who participated in the initial study reported an average of 4.94 (± 4.87) symptoms. Those reported most commonly included rhinorrhea, sneezing, headache, sore throat and dry cough. Respiratory symptoms were reported by 74%, ocular symptoms by 62%, and gastrointestinal symptoms by 35% of respondents. Residential and recreational exposures were associated with increased risks of respiratory, gastrointestinal, or ocular symptoms in univariate and adjusted multivariable analyses. Residential exposure was significantly associated with increased reporting of dry cough (p = 0.03), dyspnea (p < 0.01) and wheezy respirations (p = 0.04). Among persons reporting gastrointestinal symptoms, nausea (p = 0.02) and abdominal pain (p < 0.01) were significantly associated with residential exposure. Recreational exposure was significantly associated with sore throat and eye irritation. The findings add to the evidence that exposure to cyanobacteria at concentrations encountered during an algal bloom is associated with a diverse array of symptoms and that inhalation of aerosols constitutes an important exposure pathway.

An Assessment of Potential Threats to Human Health from Algae Blooms in the Indian River Lagoon (USA) 2018-2021: Unique Patterns of Cytotoxicity Associated with Toxins.

The Indian River Lagoon (IRL), a 156-mile-long estuary located on the eastern coast of Florida, experiences phytoplankton bloom events due to increased seasonal temperatures coupled with anthropogenic impacts. This study aimed to gather data on the toxicity to human cells and to identify secondary metabolites found in water samples collected in the IRL. Water samples from 20 sites of the IRL were collected during the wet and dry seasons over a three-year period. A panel of cell lines was used to test cytotoxicity. Hemagglutination, hemolysis, and inhibition of protein phosphatase 2A (PP2A) were also measured. Cytotoxic blooms were seen both in the south (Microcystis) and the north (Pyrodinium) of the IRL. Each toxin induced a consistent pattern of cytotoxicity in the panel of human cell lines assayed. During blooms, cytotoxicity due to a single type of toxin is obvious from this pattern. In the absence of blooms, the cytotoxicity seen reflected either a mixture of toxins or it was caused by an unidentified toxin. These observations suggest that other toxins with the potential to be harmful to human health may be present in the IRL. Moreover, the presence of toxins in the IRL is not always associated with blooms of known toxin-producing organisms.

Pseudo-nitzschia species, toxicity, and dynamics in the southern Indian River Lagoon, FL.

The Indian River Lagoon (IRL) spans approximately one-third of the east coast of Florida and, in recent years, has faced frequent harmful algal blooms (HABs). Blooms of the potentially toxic diatom, Pseudo-nitzschia, occur throughout the lagoon and were reported primarily from the northern IRL. The goal of this study was to identify species of Pseudo-nitzschia and characterize their bloom dynamics in the southern IRL system where monitoring has been less frequent. Surface water samples collected from five locations between October 2018 and May 2020 had Pseudo-nitzschia spp. present in 87% of samples at cell concentrations up to 1.9×103 cells mL-1. Concurrent environmental data showed Pseudo-nitzschia spp. were associated with relatively high salinity waters and cool temperatures. Six species of Pseudo-nitzschia were isolated, cultured, and characterized through 18S Sanger sequencing and scanning electron microscopy. All isolates demonstrated toxicity and domoic acid (DA) was present in 47% of surface water samples. We report the first known occurrence of P. micropora and P. fraudulenta in the IRL, and the first known DA production from P. micropora.

In situ digital holographic microscopy for rapid detection and monitoring of the harmful dinoflagellate, Karenia brevis.

Karenia brevis blooms, also known as red tide, are a recurring problem in the coastal Gulf of Mexico. These blooms have the capacity to inflict substantial damage to human and animal health as well as local economies. Thus, monitoring and detection of K. brevis blooms at all life stages and cell concentrations is essential for ensuring public safety. Current K. brevis monitoring methods have several limitations, including size resolution limits and concentration ranges, limited capacity for spatial and temporal profiling, and/or small sample volume processing. Here, a novel monitoring method wherein an autonomous digital holographic imaging microscope (AUTOHOLO), that overcomes these limitations and can characterize K. brevis concentrations in situ, is presented. Using the AUTOHOLO, in situ field measurements were conducted in the coastal Gulf of Mexico during an active K. brevis bloom over the 2020-21 winter season. Surface and sub-surface water samples collected during these field studies were also analyzed in the lab using benchtop holographic imaging and flow cytometry for validation. A convolutional neural network was trained for automated classification of K. brevis at all concentration ranges. The network was validated with manual counts and flow cytometry, yielding a 90% accuracy across diverse datasets with varying K. brevis concentrations. The usefulness of pairing the AUTOHOLO with a towing system was also demonstrated for characterizing particle abundance over large spatial distances, which could potentially facilitate characterization of spatial distributions of K. brevis during bloom events. Future applications of the AUTOHOLO can include integration into existing HAB monitoring networks to enhance detection capabilities for K. brevis in aquatic environments around the world.

Detection of numerous phycotoxins in young bull sharks (Carcharhinus leucas) collected from an estuary of national significance.

Florida's Indian River Lagoon (IRL) has experienced large-scale, frequent blooms of toxic harmful algae in recent decades. Sentinel, or indicator, species can provide an integrated picture of contaminants in the environment and may be useful to understanding phycotoxin prevalence in the IRL. This study evaluated the presence of phycotoxins in the IRL ecosystem by using the bull shark (Carcharhinus leucas) as a sentinel species. Concentrations of phycotoxins were measured in samples collected from 50 immature bull sharks captured in the IRL between 2018 and 2020. Ultra-performance liquid chromatography/tandem mass spectrometry was used to measure toxins in shark gut contents, plasma, and liver. Analysis of samples (n = 123) demonstrated the presence of multiple phycotoxins (microcystin, nodularin, teleocidin, cylindrospermopsin, domoic acid, okadaic acid, and brevetoxin) in 82 % of sampled bull sharks. However, most detected toxins were in low prevalence (≤25 % of samples, per sample type). This study provides valuable baseline information on presence of multiple phycotoxins in a species occupying a high trophic position in this estuary of national significance.

Dynamics of microcystins and saxitoxin in the Indian River Lagoon, Florida.

Harmful algal blooms that can produce toxins are common in the Indian River Lagoon (IRL), which covers ~250 km of Florida's east coast. The current study assessed the dynamics of microcystins and saxitoxin in six segments of the IRL: Banana River Lagoon (BRL), Mosquito Lagoon (ML), Northern IRL (NIRL), Central IRL (CIRL), Southern IRL (SIRL), and the St. Lucie Estuary (SLE). Surface water samples (n = 40) collected during the 2018 wet and 2019 dry season were analyzed to determine associations between toxins and temperature, salinity, pH, oxygen saturation, concentrations of dissolved nutrients and chlorophyll-a, presence of biosynthetic genes for toxins, relative abundance of planktonic species, and composition of the microbial community. The potential toxicity of samples was assessed using multiple mammalian cell lines. Enzyme-Linked Immunosorbent Assays were used to determine concentrations of microcystins and saxitoxin. Overall, the microcystins concentration ranged between 0.01-85.70 µg/L, and saxitoxin concentrations ranged between 0.01-2.43 µg/L across the IRL. Microcystins concentrations were 65% below the limit of quantification (0.05 µg/L), and saxitoxin concentrations were 85% below the limit of detection (0.02 µg/L). Microcystins concentrations were higher in the SLE, while saxitoxin was elevated in the NIRL and BRL. Cytotoxicity related to the presence of microcystins was seen in the SLE during the wet season. No significant patterns between cytotoxicity and saxitoxin were identified. Dissolved nutrients were identified as the most highly related parameters, explaining 53% of microcystin and 47% of saxitoxin variability. Multivariate models suggested cyanobacteria, flagellates, ciliates, and diatoms as the subset of microorganisms whose abundances were maximally correlated with saxitoxin and microcystins concentrations. Lastly, biosynthetic genes for microcystins were detected in the SLE and for saxitoxin in the BRL and NIRL. These results highlight the synergistic roles environmental and biological parameters play in influencing the dynamics of toxin production by harmful algae in the IRL.

Optical backscattering and linear polarization properties of the colony forming cyanobacterium Microcystis.

Light scattering characteristics of the cyanobacterium Microcystis are investigated with numerical models for sphere aggregates. During summer bloom seasons, Microcystis is prevalent in many inland waters across the globe. Monitoring concentrations with remote sensing techniques requires knowledge of the inherent optical properties (IOPs), especially the backscattering properties of Microcystis cells and colonies in natural settings. In situ measurements in waters dominated by Microcystis blooms have previously detected extremely high backscattering ratios, i.e., bb/b>0.043 at 443 nm [1], the highest to our knowledge in the natural environment. These highbb/bvalues could hold promise as a diagnostic tool in identifying and monitoring Microcystis using optical approaches. However, it has been unclear how this type of optically 'soft' organic particle can generate such highbb/bvalues. In this study, the Multiple Sphere T-matrix (MSTM) model is used to calculate the IOPs of model colonies, including bb/b. Colony sizes in the model ranged from several cells to several hundred and both colony packing density and cell gas vacuole content were varied. Results are compared with model results for equivalent-volume spheres (EVS) and direct in situ measurements. Colony formation was required in the modeling to reproduce the high bb/bconsistent with in situ measurements. The combination of moderate to very dense colony (packing density >30%) and high gas vacuole content in individual cells (volume percentage >20%) was the most favorable condition leading to rapid increases in bb/bwith increasing number of cells Ncell of the colony. Significant linear correlations were observed betweenbb/b and Ncell1/3 for these colonies, wherebb/b increased beyond 0.04 once cell number reached about 1000 cells in the case with the most densely packed cells and highest gas vacuole content. Within commonly observed colony sizes (Ncell <106), colonies with high gas vacuole content exhibited bb/bvalues up to 0.055, consistent with direct measurements from Lake Erie. Polarized scattering was also of interest as a diagnostic tool, particularly with future Earth-orbiting polarimeters being deployed for the NASA Plankton, Aerosols, Cloud, ocean Ecosystem (PACE) mission. The Degree of Linear Polarization (DoLP), expressed by the ratio of two Mueller matrix elements-P12/P11, decreased with increasing colony cell number for Microcystis. Another ratio of two Mueller matrix elementsP22/P11, an index for nonsphericity, also decreased with increasing colony size. In addition to higher relative backscattering, greater colony packing density and larger gas vacuole sizes both led to lower DoLP peak magnitude and lowerP22/P11. An optical opposition feature due to constructive phase interference that was observed previously for cosmic dusts is also present for these modeled colonies, manifested by a narrow intensity peak and negative polarization dip near exact backscattering direction, gradually forming as colony size increases.

Exposure to microcystin among coastal residents during a cyanobacteria bloom in Florida.

Florida has experienced multiple cyanobacteria blooms in recent years the most severe of which occurred in 2016 and 2018. Several toxins are produced by proliferating cyanobacteria, including the hepatotoxin microcystin (MC). Harmful algal blooms (HABs) caused by cyanobacteria have the potential to impact public health. However, to date there have been limited attempts to quantify exposure in human populations. This study investigated potential exposure to the cyanobacterial toxin, MC by measuring concentrations in swabs of the nasal mucosa. In addition, the relationships between nasal concentrations of MC, environmental concentrations and activity patterns were assessed. Participants (n = 125) were recruited in 2018 during a cyanobacterial bloom of Microcystis aeruginosa and completed a questionnaire which included location, type, and duration of recreational or occupational contact with impacted waterways within the last 10 days. Water samples were collected concurrently. A sterile swab was used to collect a sample from the nasal mucosa. Concentrations of MC were measured by ELISA. Of the 121 participants who provided nasal swabs, 115 (95.0 %) had concentrations of MC above the limit of detection with a mean concentration of 0.61 + 0.75 ppb. There were significant differences (p < 0.01) in mean MC concentration between individuals with direct contact with impacted waters (0.77 + 0.88 ppb) compared to those with no recent contact (0.37 + 0.49 ppb). Higher concentrations were observed among occupationally exposed individuals. Nasal concentrations of MC varied significantly over time and location of exposure to the bloom, concordant with concentrations in water samples. The results suggest that inhalation of aerosols may be an important pathway for exposure to MC. Nasal MC concentrations were generally highest during periods when concentrations in the surrounding waters peaked. Further research is needed to characterize the public health implications of exposure to cyanobacterial blooms.

Evidence for ubiquitous preferential particle orientation in representative oceanic shear flows.

In situ measurements were undertaken to characterize particle fields in undisturbed oceanic environments. Simultaneous, co-located depth profiles of particle fields and flow characteristics were recorded using a submersible holographic imaging system and an acoustic Doppler velocimeter, under different flow conditions and varying particle concentration loads, typical of those found in coastal oceans and lakes. Nearly one million particles with major axis lengths ranging from ∼14 µm to 11.6 mm, representing diverse shapes, sizes, and aspect ratios were characterized as part of this study. The particle field consisted of marine snow, detrital matter, and phytoplankton, including colonial diatoms, which sometimes formed "thin layers" of high particle abundance. Clear evidence of preferential alignment of particles was seen at all sampling stations, where the orientation probability density function (PDF) peaked at near horizontal angles and coincided with regions of low velocity shear and weak turbulent dissipation rates. Furthermore, PDF values increased with increasing particle aspect ratios, in excellent agreement with models of spheroidal particle motion in simple shear flows. To the best of our knowledge, although preferential particle orientation in the ocean has been reported in two prior cases, our findings represent the first comprehensive field study examining this phenomenon. Evidence of nonrandom particle alignment in aquatic systems has significant consequences to aquatic optics theory and remote sensing, where perfectly random particle orientation and thus isotropic symmetry in optical parameters is assumed. Ecologically, chain-forming phytoplankton may have evolved to form large aspect ratio chains as a strategy to optimize light harvesting.

Ploidy Distribution of the Harmful Bloom Forming Macroalgae Ulva spp. in Narragansett Bay, Rhode Island, USA, Using Flow Cytometry Methods.

Macroalgal blooms occur worldwide and have the potential to cause severe ecological and economic damage. Narragansett Bay, RI is a eutrophic system that experiences summer macroalgal blooms composed mostly of Ulva compressa and Ulva rigida, which have biphasic life cycles with separate haploid and diploid phases. In this study, we used flow cytometry to assess ploidy levels of U. compressa and U. rigida populations from five sites in Narragansett Bay, RI, USA, to assess the relative contribution of both phases to bloom formation. Both haploid gametophytes and diploid sporophytes were present for both species. Sites ranged from a relative overabundance of gametophytes to a relative overabundance of sporophytes, compared to the null model prediction of √2 gametophytes: 1 sporophyte. We found significant differences in cell area between ploidy levels for each species, with sporophyte cells significantly larger than gametophyte cells in U. compressa and U. rigida. We found no differences in relative growth rate between ploidy levels for each species. Our results indicate the presence of both phases of each of the two dominant bloom forming species throughout the bloom season, and represent one of the first studies of in situ Ulva life cycle dynamics.