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12 pages, 11234 KiB  
Article
Advancements in the Programmable Hyperspectral Seawater Scanner Measurement Technology for Enhanced Detection of Harmful Algal Blooms
by John J. Langan and Jungyun Bae
J. Mar. Sci. Eng. 2024, 12(10), 1746; https://doi.org/10.3390/jmse12101746 - 3 Oct 2024
Viewed by 788
Abstract
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 [...] Read more.
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
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Figure 1

Figure 1
<p>The location of the PHySS at New Pass, shown on Google Earth.</p>
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<p>The fourth derivative absorption spectra of the phytoplankton model of <span class="html-italic">Karenia brevis</span> highlighting distinctive features used for species identification.</p>
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<p>The fourth derivative spectrum of the phytoplankton library used in the operating system.</p>
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<p>The Similarity Index over time.</p>
Full article ">Figure 5
<p>A graphical overlay of the SI against the cell count.</p>
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<p>The Similarity Index Q-Q plot.</p>
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<p>The physical cell count Q-Q plot.</p>
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<p>A graphical overlay showing the elevated SI while the cell counts are zero.</p>
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19 pages, 5139 KiB  
Article
Plankton Community Changes and Nutrient Dynamics Associated with Blooms of the Pelagic Cyanobacterium Trichodesmium in the Gulf of Mexico and the Great Barrier Reef
by Judith M. O’Neil, Cynthia A. Heil, Patricia M. Glibert, Caroline M. Solomon, Joan Greenwood and Jack G. Greenwood
Water 2024, 16(12), 1663; https://doi.org/10.3390/w16121663 - 11 Jun 2024
Viewed by 1275
Abstract
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 [...] Read more.
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
(This article belongs to the Section Biodiversity and Functionality of Aquatic Ecosystems)
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Figure 1

Figure 1
<p><span class="html-italic">Trichodesmium</span> blooms. (<b>a</b>) Surface slicks of <span class="html-italic">T. erythraeum</span> present during the early stages of bloom development, with Heron Island in the background; (<b>b</b>) degenerating surface slicks of <span class="html-italic">T. erythraeum</span> during the later stage of a bloom; (<b>c</b>) <span class="html-italic">T. erythraeum</span> slicks deposited on coral and rock along the edge of Heron Island during the later stages of a bloom; (<b>d</b>) surface slicks of <span class="html-italic">Trichodesmium</span> spp. in the Gulf of Mexico; and (<b>e</b>) <span class="html-italic">Trichodesmium</span> spp. accumulations reaching the beaches of Florida. Photos (<b>a</b>–<b>c</b>) by P. Glibert. Photo (<b>d</b>) by Sara Klass, Mote Marine Laboratory, and photo (<b>e</b>) by C. Heil.</p>
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<p>Occurrence of <span class="html-italic">Trichodesmium</span> spp. in Florida HAB monitoring samples in the eastern Gulf of Mexico from 2007 to 2024. Data from Florida Fish and Wildlife Conservation Commission, Fish and Wildlife Research Institute (FWC-FWRI) HAB Monitoring Database.</p>
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<p>Relationship between <span class="html-italic">Trichodesmium</span> spp. and water temperature in the eastern Gulf of Mexico from 2007 to the present. Data from Florida Fish and Wildlife Conservation Commission, Fish and Wildlife Research Institute (FWC-FWRI) HAB Monitoring Database.</p>
Full article ">Figure 4
<p>Log <span class="html-italic">Trichodesmium</span> (cell L<sup>−1</sup>) and bloom severity index of <span class="html-italic">Karenia brevis</span> (for blooms with abundances &gt;100,000 cells L<sup>−1</sup>). Data encompass the time period 2007–2019. All data are averaged monthly. Bloom severity index data obtained from Stumpf et al. [<a href="#B26-water-16-01663" class="html-bibr">26</a>].</p>
Full article ">Figure 5
<p><span class="html-italic">Trichodesmium erythraeum</span> abundance (colonies L<sup>−1</sup>) off Heron Island over the course of the study (bar graph). Values are the mean of two replicates (±S.D.). Concentrations (μM, ±S.D.) of NO<sub>x</sub> (Nitrate + Nitrite; hatched circle); NH<sub>4</sub><sup>+</sup> (black circle); PO<sub>4</sub><sup>3−</sup> (white circle), and dissolved organic nitrogen (grey circle) at the time of each sampling of the eularian study.</p>
Full article ">Figure 6
<p>Changes (±S.D.) in the percent contribution of (<b>A</b>) diatoms and (<b>B</b>) dinoflagellates to the phytoplankton community inside (black bars) and outside (grey bars) of the <span class="html-italic">Trichodesmium</span> bloom. All values exclude <span class="html-italic">Trichodesmium erythraeum</span> from total phytoplankton abundance. Differences within and outside of the <span class="html-italic">T. erythraeum</span> bloom were apparent by the second day of the study. (A statistically significant difference (&lt;0.05) between values is indicated by the presence of an *).</p>
Full article ">Figure 7
<p>Microscopic image of a dividing cell of the dinoflagellate <span class="html-italic">Dinophysis caudata</span> and a single <span class="html-italic">Trichodesmium erythraeum</span> trichome, from the Great Barrier Reef Lagoon. <span class="html-italic">Dinophysis caudata</span> was one of the species of dinoflagellates that increased in abundance in relation to <span class="html-italic">T. erythraeum</span> colony increase (Photo: C. Heil).</p>
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<p>(<b>A</b>) Top panel; Abundance of the copepod <span class="html-italic">Macrosetella gracilis</span> (individuals m<sup>−3</sup>) within the <span class="html-italic">T. erythraeum</span> bloom during the course of the study. (<b>B</b>) Bottom panel; Life stage (nauplii, copepodite, or adult) of <span class="html-italic">M. gracilis</span> population as percentage of total number of individuals counted over course of the bloom. (N.B.: On Day 7, no sampling occurred due to a front that passed through the study area, with increased wind speed increases, which curtailed boating.)</p>
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<p><span class="html-italic">Trichodesmium erythraeum</span> ingestion rates (μg C μg copepod C<sup>−1</sup> h<sup>−1</sup>) by adult <span class="html-italic">Macrosetella gracilis</span> inside (black circles) and outside (black squares) of a <span class="html-italic">T. erythraeum</span> bloom measured on Heron Island, Great Barrier Reef. Each value is the mean of 3–4 replicates (±standard error).</p>
Full article ">Figure 10
<p>(<b>a</b>) A nauplius of the copepod <span class="html-italic">Macrosetella gracilis</span> on a <span class="html-italic">Trichodesmium</span> colony; (<b>b</b>) another <span class="html-italic">M. gracilis</span> between the filament of a tuft colony of <span class="html-italic">Trichodesmium</span>; (<b>c</b>) an adult <span class="html-italic">M. gracilis</span> with a puff colony of <span class="html-italic">T. thiebautii</span>. Photos by J.M. O’Neil.</p>
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14 pages, 5249 KiB  
Article
Temperature-Dependent Mixotrophy in Natural Populations of the Toxic Dinoflagellate Karenia brevis
by So Hyun Ahn and Patricia M. Glibert
Water 2024, 16(11), 1555; https://doi.org/10.3390/w16111555 - 29 May 2024
Cited by 1 | Viewed by 873
Abstract
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 [...] Read more.
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 (rETRmax), 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 rETRmax of Synechococcus controls generally increased over time with a more substantial increase at warmer temperatures, but, in the presence of grazers, the rETRmax 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
(This article belongs to the Section Biodiversity and Functionality of Aquatic Ecosystems)
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Figure 1

Figure 1
<p>Experimental design examining short-term temperature effects on grazing rates by <span class="html-italic">K. brevis</span> on <span class="html-italic">Synechococcus</span>. SD, BD, R<sub>1,</sub> and R<sub>2</sub> represent South Dock, Bay Dock, and ratios 1 and 2, respectively. Prey-only treatments are shown in green, <span class="html-italic">K. brevis</span>-only treatments are shown in orange, and mixed predator + prey treatments are shown in purple. Detailed information is presented in <a href="#water-16-01555-t002" class="html-table">Table 2</a>.</p>
Full article ">Figure 2
<p>The changes of <span class="html-italic">Synechococcus</span> abundance with or without grazers at different temperatures and ratios in (<b>A</b>) Experiment 1 SD, (<b>B</b>) Experiment 2 SD, (<b>C</b>) Experiment 3 BD, (<b>D</b>) Experiment 3 SD, and (<b>E</b>) Experiment 4 SD treatments. Solid lines with empty symbols represent controls of <span class="html-italic">Synechococcus</span>. In contrast, dotted lines with filled symbols represent <span class="html-italic">Synechococcus</span> mixed with <span class="html-italic">K. brevis</span> bloom waters from the South Dock (SD) and Bay Dock (BD): diamonds (<b>♦</b>, ♢) for the treatments at 19 °C, circles (○, ●) for the treatments at 24 °C, and triangles (△, ▲) for the treatments at 29 °C. Note that treatments in Experiment 1 and 2 include 2 prey-to-grazer ratios (R<sub>1</sub>, R<sub>2</sub>) and 2 temperatures (24, 29 °C), while those in Experiment 3 and 4 include 1 prey-to-grazer ratio (R<sub>1</sub>) and 3 temperatures (19, 24, and 29 °C).</p>
Full article ">Figure 3
<p>The changes in <span class="html-italic">Karenia brevis</span> abundance at different temperatures and ratios in (<b>A</b>) Experiment 1 SD, (<b>B</b>) Experiment 2 SD, (<b>C</b>) Experiment 3 BD, (<b>D</b>) Experiment 3 SD, and (<b>E</b>) Experiment 4 SD treatments. R<sub>1</sub>, R<sub>2</sub>, SD, and BD represent ratio 1, ratio 2, South Dock, and Bay Dock, respectively. The scales of the X and Y axis are different for different experiments. Note that treatments in Experiment 1 and 2 include 2 prey-to-grazer ratios (R<sub>1</sub>, R<sub>2</sub>) and 2 temperatures (24, 29 °C) while those in Experiment 3 and 4 include 1 prey-to-grazer ratio (R<sub>1</sub>) and 3 temperatures (19, 24, 29 °C).</p>
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<p>The maximum (<b>A</b>) grazing coefficient (<span class="html-italic">g</span>), (<b>B</b>) clearance rate, and (<b>C</b>) Ingestion rate of <span class="html-italic">K. brevis</span> on <span class="html-italic">Synechococcus</span> at 19, 24, and 29 °C. R<sub>1</sub>, R<sub>2</sub>, SD, and BD represent ratio 1, ratio 2, South Dock, and Bay Dock, respectively.</p>
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<p>The relationship between daily ingestion rate by <span class="html-italic">K. brevis</span> on <span class="html-italic">Synechococcus</span> and (<b>A</b>) the prey-to-grazer ratio and temperature and (<b>B</b>) prey concentrations and temperature. The multiple linear regression plane and R<sup>2</sup> are shown.</p>
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<p>The maximum relative electron transfer rate, rETR<sub>max</sub>, of <span class="html-italic">Synechococcus</span> over time in Experiments 1 (<b>A</b>–<b>F</b>) and 2 (<b>G</b>–<b>N</b>). R<sub>1</sub>, R<sub>2</sub>, SD, and BD represent ratio 1, ratio 2, South Dock, and Bay Dock, respectively.</p>
Full article ">Figure 7
<p>The maximum relative electron transfer rate, rETR<sub>max</sub>, of <span class="html-italic">Synechococcus</span> over time in Experiments 3 SD (<b>A</b>–<b>F</b>) and BD (<b>G</b>–<b>L</b>) and Experiment 4 SD (<b>M</b>–<b>R</b>). SD, and BD represent South Dock, and Bay Dock, respectively.</p>
Full article ">Figure 8
<p>The maximum relative electron transfer rate, rETR<sub>max</sub>, of <span class="html-italic">Karenia brevis</span> over time in Experiments 1 (<b>A</b>–<b>F</b>) and 2 (<b>G</b>–<b>L</b>). R<sub>1</sub>, R<sub>2</sub>, SD, and BD represent ratio 1, ratio 2, South Dock, and Bay Dock, respectively.</p>
Full article ">Figure 9
<p>The maximum relative electron transfer rate, rETR<sub>max</sub>, of <span class="html-italic">Karenia brevis</span> over time in Experiment 3 SD (<b>A</b>–<b>F</b>), Experiment 3 BD (<b>G</b>–<b>L</b>) and Experiment 4 SD (<b>M</b>–<b>R</b>). R<sub>1</sub>, R<sub>2</sub>, SD, and BD represent ratio 1, ratio 2, South Dock, and Bay Dock, respectively.</p>
Full article ">
15 pages, 2534 KiB  
Article
Mitigation of Karenia brevis Cells and Brevetoxins Using Curcumin, a Natural Supplement
by Emily R. Hall, Cynthia A. Heil, Jessica D. Frankle, Sarah Klass, Victoria Devillier, Vincent Lovko, Jennifer H. Toyoda and Richard Pierce
Water 2024, 16(10), 1458; https://doi.org/10.3390/w16101458 - 20 May 2024
Viewed by 1382
Abstract
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 [...] Read more.
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 article belongs to the Special Issue Eutrophication and Harmful Algae in Aquatic Ecosystems)
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Figure 1

Figure 1
<p>Percent reduction (%R) of low-dose (0.1–5 mg/L) curcumin treatments on <span class="html-italic">K. brevis</span> cell abundance (cells/L) relative to control. Lines represent mean ± 95% CI from additive GLM of time and treatment as explanatory variables. N varies by treatment and time point, N = 3 at minimum.</p>
Full article ">Figure 2
<p>Percent reduction (%R) of high-dose (10–40 mg/L) curcumin treatments on <span class="html-italic">K. brevis</span> cell abundance (cells/L) relative to control. Lines represent mean ± 95% CI from additive GLM of time and treatment as explanatory variables. N varies by treatment and time point, N = 3 at minimum.</p>
Full article ">Figure 3
<p>Percent reduction (%R) of low-dose (1–5 mg/L) curcumin treatments on <span class="html-italic">K. brevis</span> total toxins relative to control. Lines represent mean ± 95% CI from additive GLM of time and treatment as explanatory variables. Total toxin measurements are sums of brevetoxin analogs BTX-1, BTX-2, BTX-3, and BTX-B5 in ng/L. N varies by treatment and time point, N = 3 at minimum.</p>
Full article ">Figure 4
<p>Percent reduction (%R) of high-dose (10–40 mg/L) curcumin treatments on <span class="html-italic">K. brevis</span> total toxins relative to control. Lines represent mean ± 95% CI from additive GLM of time and treatment as explanatory variables. Total toxin measurements are sums of brevetoxin analogs BTX-1, BTX-2, BTX-3, and BTX-B5 in ng/L. N varies by treatment and time point, N = 2 at minimum.</p>
Full article ">Figure 5
<p>Total brevetoxin concentrations (ng/L) in low (1–5 mg/L)-dose treatments over time divided by congener: BTX-1, BTX-2, BTX-3, and BTX-B5. In the x-axis, C indicates control treatment (no curcumin addition), while T indicates curcumin treatment, with the number of hours post addition indicated after the letter. N varies by treatment and time point, N = 3 at minimum.</p>
Full article ">Figure 6
<p>Total brevetoxin concentrations (ng/L) in high (10–40 mg/L)-dose treatments over time divided by congener: BTX-1, BTX-2, BTX-3, and BTX-B5. In the x-axis, C indicates control treatment (no curcumin addition), while T indicates curcumin treatment, with the number of hours post addition indicated after the letter. N varies by treatment and time point, N = 2 at minimum.</p>
Full article ">
24 pages, 8090 KiB  
Article
Karenia brevis and Pyrodinium bahamense Utilization of Dissolved Organic Matter in Urban Stormwater Runoff and Rainfall Entering Tampa Bay, Florida
by Amanda L. Muni-Morgan, Mary G. Lusk and Cynthia A. Heil
Water 2024, 16(10), 1448; https://doi.org/10.3390/w16101448 - 19 May 2024
Cited by 3 | Viewed by 1524
Abstract
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 [...] Read more.
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 article belongs to the Special Issue Eutrophication and Harmful Algae in Aquatic Ecosystems)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>A map of the stormwater runoff and rainfall collection sites in Pinellas County, FL, for water used as inocula for the bioassays. The blue raindrop icon represents the rainfall collection site. Red (Beachside and Mainland) and black (Clearwater and Safety Harbor) colored icons represent sites used to collect stormwater runoff for the <span class="html-italic">K. brevis</span> and <span class="html-italic">P. bahamense</span> bioassays, respectively. The red square inset on the state of Florida to the right corresponds to the Tampa Bay region.</p>
Full article ">Figure 2
<p>Fluorescence signatures of PARAFAC modeled BIO-MOD components from the <span class="html-italic">K. brevis</span> and <span class="html-italic">P. bahamense</span> bioassays. The EEM contour plots (<b>left panel</b>) for each component are displayed with emission wavelengths (nm) on the Y-axis, excitation wavelengths (nm) on the X-axis, and a colored scale bar representing fluorescence intensity in quinine sulfate units (QSU) from low (blue) to high (yellow). The corresponding excitation/emission spectra for each component are placed to the (<b>right</b>). The dashed/solid lines represent excitation/emission spectra wavelengths (nm), respectively, with the Y-axis representing fluorescence intensity in QSU.</p>
Full article ">Figure 3
<p>Fluorescence signatures of PARAFAC modeled SW-MOD components from the source water used as the inoculum for the <span class="html-italic">K. brevis</span> and <span class="html-italic">P. bahamense</span> bioassays. The EEM contour plots (<b>left panel</b>) for each component are displayed with emission wavelengths (nm) on the Y-axis, excitation wavelengths (nm) on the X-axis, and a colored scale bar representing fluorescence intensity in quinine sulfate units (QSU) from low (blue) to high (yellow). The corresponding excitation/emission spectra for each component are placed to the <b>right</b>. The dashed/solid lines represent excitation/emission spectra wavelengths (nm), respectively, with the Y-axis representing fluorescence intensity in QSU.</p>
Full article ">Figure 4
<p>Changes in the relative fluorescence of FDOM components for the <span class="html-italic">K. brevis</span> bioassay. The colored bars represent individual components generated from the PARAFAC model, with comparisons between initial and final sampling timepoints for each treatment on the X-axis. The error bars represent the standard error of the mean. Relative fluorescence is expressed in quinine sulfate units (QSU).</p>
Full article ">Figure 5
<p>Changes in the relative fluorescence of FDOM components for the <span class="html-italic">P. bahamense</span> bioassay. The colored bars represent individual components, with comparisons between initial and final sampling timepoints for each treatment on the X-axis. The error bars represent the standard error of the mean. Relative fluorescence is expressed in quinine sulfate units (QSU).</p>
Full article ">Figure 6
<p>Concentrations of nitrogen forms (dissolved organic nitrogen = DON) and dissolved organic carbon (DOC) for the stormwater and rainfall used as inocula for both bioassays. Each stormwater or rainfall treatment collected for the respective bioassay is defined on the X-axis. Note the differences in axes and scales.</p>
Full article ">Figure 7
<p>Change in concentrations of nitrogen forms (dissolved organic nitrogen = DON) and dissolved organic carbon (DOC) vs. time for the <span class="html-italic">K. brevis</span> bioassay. Colored lines represent individual rainfall or stormwater treatments. Error bars represent standard error of mean. Note differences in Y-axes.</p>
Full article ">Figure 8
<p>Change in concentrations of nitrogen forms (dissolved organic nitrogen = DON) and dissolved organic carbon (DOC) vs. time for the <span class="html-italic">P. bahamense</span> bioassay. Colored lines represent individual rainfall or stormwater treatments. Error bars represent standard error of mean. Note differences in Y-axes.</p>
Full article ">Figure 9
<p>Relative fluorescence of components for stormwater runoff and rainfall used as inocula for <span class="html-italic">K. brevis</span> (KB) and <span class="html-italic">P. bahamense</span> (PB) bioassays. Colored bars represent individual components generated from PARAFAC model. SW1 represents terrestrial humic-like FDOM. Each stormwater or rainfall treatment collected for the respective bioassay is defined on the X-axis. Relative fluorescence is expressed in quinine sulfate units (QSU).</p>
Full article ">Figure 10
<p><span class="html-italic">Karenia brevis</span> cell concentrations vs. time in hours. Colored lines represent individual rainfall or stormwater treatments. Error bars represent standard error of mean. Colored asterisk denotes statistical significance of maximum cell concentrations vs. Control treatment (<span class="html-italic">p</span> ≤ 0.05).</p>
Full article ">Figure 11
<p><span class="html-italic">Pyrodinium bahamense</span> cell concentrations vs. time in hours. Colored lines represent individual rainfall or stormwater treatments. Error bars represent standard error of mean. Colored asterisk denotes statistical significance of maximum cell concentrations vs. Control treatment (<span class="html-italic">p</span> ≤ 0.05).</p>
Full article ">
25 pages, 402 KiB  
Review
Emerging Insights into Brevetoxicosis in Sea Turtles
by Remco A. Nederlof, Dion van der Veen, Justin R. Perrault, Robin Bast, Heather W. Barron and Jaco Bakker
Animals 2024, 14(7), 991; https://doi.org/10.3390/ani14070991 - 22 Mar 2024
Viewed by 1971
Abstract
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 [...] Read more.
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
(This article belongs to the Special Issue Clinical Pharmacology and Toxicology for Wildlife)
15 pages, 4213 KiB  
Article
Karenia brevis Extract Induces Cellular Entry through Distinct Mechanisms in Phagocytic RAW 264.7 Macrophages versus Non-Phagocytic Vero Cells
by Laurie A. Minns, Kathryn T. Sausman, Ariel P. Brown, Robert A. York and Jennifer R. McCall
Mar. Drugs 2024, 22(1), 4; https://doi.org/10.3390/md22010004 - 19 Dec 2023
Viewed by 4152
Abstract
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, [...] Read more.
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 ? and pheophytin ?. 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 ? and pheophytin ?, 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
(This article belongs to the Section Marine Pharmacology)
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<p><span class="html-italic">K. brevis</span> algal extracts absorb and emit fluorescence at multiple wavelengths. Spectral properties of <span class="html-italic">K. brevis</span> extract at 1 mg/mL. Extracts absorbed light with peak wavelengths at 405 nm and 664 nm (panel (<b>A</b>)). Extracts were then excited with peak wavelengths (405 nm: panel (<b>B</b>) or 664 nm: panel (<b>C</b>)).</p>
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<p>HPLC-UV chromatogram with detection wavelength at 667 nm of a chlorophyll α reference standard (panel (<b>A</b>)) and <span class="html-italic">K. brevis</span> algae extract (panel (<b>B</b>)). The chromatogram for the algae extract indicates that although chlorophyll α was present, pheophytin α was most abundant. Mass spectroscopy later identified Pheophytin α as the large peak in panel (<b>B</b>).</p>
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<p>Fluorescence accumulation in RAW 264.7 cells (Panel (<b>A</b>)) and Vero cells (Panel (<b>B</b>)) over a 30 min to 48 h time course as excited by violet laser and read on Brilliant Violet 650 filter. Symbols indicate a statistically significant difference from no treatment (*), 0.5 h (+), 2 h (#), and 24 h (@) with <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>RAW 264.7 macrophages accumulate more <span class="html-italic">K. brevis</span> extract fluorescence in the presence of EGA, a mem-brane trafficking inhibitor (Panel (<b>A</b>)), whereas Vero cells display a decrease in fluorescence in a dose-dependent manner with EGA pre-treatment (Panel (<b>B</b>)). RAW 264.7 showed no change in fluorescence accumulation in the presence of Dynasore, a dynamin-dependent inhibitor (Panel (<b>C</b>)). Non-phagocytotic Vero cells show an increase in extract fluores-cence uptake in a dose-dependent manner when treated with Dynasore (Panel (<b>D</b>)). * <span class="html-italic">p</span> &lt; 0.05 when compared to vehicle control.</p>
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<p>RAW 264.7 (Panel (<b>A</b>,<b>B</b>)) and Vero cells (Panel (<b>C</b>,<b>D</b>)) were pretreated with EGA for one hour (Panel (<b>B</b>,<b>D</b>)) or not (Panel (<b>A</b>,<b>C</b>)), then treated with fluorescent components of <span class="html-italic">K. brevis</span>, the extract, chlorophyll α, and pheophytin α (Cy5 pink color) for approximately 24 h. Cells were then treated with nucleus Hoechst stain (DAPI blue color) for 1 h, washed, and imaged using Image Xpress PICO at 20×.</p>
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<p>Assessment of macrophage alternative activation states by cytokine secretion ((<b>A</b>) n-3) IL-10, TNF α and IL-6 and cell surface expression (<b>B</b>) of CD206 (n-10), CD80 (n-3) and IL4Rα (n-3). RAW 264.7 macrophages were treated with algal extracts for 24 h and stained with fluorescent antibodies prior to analysis on a flow cytometer. * indicates a statistically significant difference from vehicle control (VC) with <span class="html-italic">p</span> &lt; 0.05.</p>
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19 pages, 2499 KiB  
Article
Acute Effects of Brevetoxin-3 Administered via Oral Gavage to Mice
by Peggy Barbe, Jordi Molgó, Robert Thai, Apolline Urman, Denis Servent, Nathalie Arnich and Mathilde Keck
Mar. Drugs 2023, 21(12), 644; https://doi.org/10.3390/md21120644 - 16 Dec 2023
Viewed by 2283
Abstract
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 [...] Read more.
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
(This article belongs to the Special Issue Emerging Toxins Accumulation in Shellfish)
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<p>Swiss mouse body weight (<b>a</b>) and gain or loss of weight (<b>b</b>) after BTX-3 oral administration for each group (BTX doses are indicated, female are represented in red striped histograms) over 48 h. Mixed-effects model followed by Dunnett’s multiple comparisons test was used; * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; <span class="html-italic">n</span> = 5 mice/group, except for male at 1,500 µg kg<sup>−1</sup> bw, where three mice were euthanized at T<sub>4h</sub>.</p>
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<p>Swiss mouse temperature (<b>a</b>) and gain or loss of temperature (<b>b</b>) after BTX-3 oral administration for each group (BTX doses are indicated, female are represented in red striped histograms) over 48 h. Mixed-effects model followed by Dunnett’s multiple comparisons test was used; * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; <span class="html-italic">n</span> = 5 mice/group, except for male at 1,500 µg kg<sup>−1</sup> bw, where three mice were euthanized at T<sub>4h</sub>.</p>
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<p>Swiss mouse success or failure in grip test (<b>a</b>), muscle activity (<b>b</b>), jaw movements indicating difficulty to breath (<b>c</b>), and tremors (<b>d</b>) observed after BTX-3 oral administration for each group (BTX doses are indicated, female are represented in red striped histograms) over 48 h. A score was attributed to symptomatology, with grade 1 corresponding to normal symptomatology, grade 2 to moderate modifications in symptomatology, and grade 3 to important modifications in symptomatology. No statistical tests could be applied; <span class="html-italic">n</span> = 5 mice/group, except for male at 1,500 µg kg<sup>−1</sup> bw, where three mice were euthanized at T<sub>4h</sub>.</p>
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<p>Schematic representation of the experimental procedure consisting of acute oral gavage administration with purified BTX-3 to male and female mice, followed by scoring at the three indicated time periods.</p>
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<p>Calibration curve for dosage of PbTx-3.</p>
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<p>Schematic representation of the pilot study consisting of acute oral gavage administration (just after scoring) with purified BTX-3 to female mice, followed by observation for 4 h, scoring at 2 h, and scoring from day 1 (D<sub>1</sub>) to day 14 (D<sub>14</sub>).</p>
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<p>Clinical chemistry in the female Swiss mouse analyzed 14 days after BTX-3 administration in each group studied (doses are indicated). Thresholds (min and max) for mice administered only with the vehicle used to dissolve BTX-3 are indicated by the dashed lines (in grey). The Kruskal–Wallis test followed by Dunn’s multiple comparisons test was used; non-significant effects observed; <span class="html-italic">n</span> = 2 mice/group.</p>
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27 pages, 699 KiB  
Review
Metabolomics Approach to Reveal the Effects of Ocean Acidification on the Toxicity of Harmful Microalgae: A Review of the Literature
by Tsz-Ki Victoria Tsui and Hang-Kin Kong
AppliedChem 2023, 3(1), 169-195; https://doi.org/10.3390/appliedchem3010012 - 16 Mar 2023
Cited by 3 | Viewed by 3165
Abstract
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 [...] Read more.
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
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<p>Schematic diagram illustrating the use of metabolomics to elucidate the individual effects of confounding factors on the carbon allocation of microalgae.</p>
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9 pages, 721 KiB  
Communication
Insights into Stress-Induced Death Processes during Aging in the Marine Bloom-Forming Dinoflagellate Karenia brevis
by Yida Gao and Deana L. Erdner
J. Mar. Sci. Eng. 2022, 10(12), 1993; https://doi.org/10.3390/jmse10121993 - 14 Dec 2022
Cited by 1 | Viewed by 1364
Abstract
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 [...] Read more.
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
(This article belongs to the Special Issue Marine Harmful Algae)
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<p>Cellular stress responses in <span class="html-italic">K. brevis</span> cells exposed to 180 μM H<sub>2</sub>O<sub>2</sub> (circles) and their corresponding control cultures (triangles) at early-log phase (<b>A</b>,<b>D</b>,<b>G</b>,<b>J</b>), mid-log phase (<b>B</b>,<b>E</b>,<b>H</b>,<b>K</b>) and stationary phase (<b>C</b>,<b>F</b>,<b>I</b>,<b>L</b>). (<b>A</b>–<b>C</b>) Proportion of cells showing reactive oxygen species (ROS). (<b>D</b>–<b>F</b>) Proportion of cells showing caspase-like activity. (<b>G</b>–<b>I</b>) Proportion of cells stained with SYTOX. (<b>J</b>–<b>L</b>) Live cell density. Data points show average values obtained from triplicate cultures, and error bars show standard deviation of the replicates (<span class="html-italic">n</span> = 6). Asterisks indicate a significant difference between treatments and controls (<span class="html-italic">p</span> &lt; 0.05).</p>
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13 pages, 1704 KiB  
Article
The Effects of the Harmful Algal Bloom Species Karenia brevis on Survival of Red Porgy (Pagrus pagrus) Larvae
by Richard Wayne Litaker, Alex K. Bogdanoff, Donnie Ransom Hardison, William C. Holland, Andrew Ostrowski and James A. Morris
Toxins 2022, 14(7), 439; https://doi.org/10.3390/toxins14070439 - 28 Jun 2022
Cited by 4 | Viewed by 2518
Abstract
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 [...] Read more.
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 L1, 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
(This article belongs to the Special Issue Effects of Harmful Algal Blooms on Aquatic Organisms)
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<p>Red porgy larvae percent survival when exposed to the three <span class="html-italic">K. brevis</span> strains tested and produced either low, medium, or high concentrations of brevetoxin. Error bars are standard deviations of the mean. (<b>A</b>) Strain SP1 which contained trace amounts of pg BTx-1, 2, 3 cell<sup>−1</sup>. (<b>B</b>) Strain NCMA2228 averaged 9.9 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2011 and 10.9 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2012. (<b>C</b>) Strain SP3 averaged 13.5 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2011 and 14.5 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2012.</p>
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<p>Red porgy larvae percent survival when exposed to the three <span class="html-italic">K. brevis</span> strains tested and produced either low, medium, or high concentrations of brevetoxin. Error bars are standard deviations of the mean. (<b>A</b>) Strain SP1 which contained trace amounts of pg BTx-1, 2, 3 cell<sup>−1</sup>. (<b>B</b>) Strain NCMA2228 averaged 9.9 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2011 and 10.9 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2012. (<b>C</b>) Strain SP3 averaged 13.5 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2011 and 14.5 pg BTx-1, 2, 3 cell<sup>−1</sup> in 2012.</p>
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<p>Dose-response curve created using combined data for all the 24 h experiments where red porgy larvae were exposed to varying cell concentrations of the three <span class="html-italic">K. brevis</span> strains tested. The black dots represent the mean survivorship for the different days post hatch larvae exposed to a given concentration of <span class="html-italic">Karenia brevis</span> ± 1 SD. The resulting estimated EC<sub>50</sub> for the red porgy larvae was ~163,000 <span class="html-italic">K. brevis</span> cells L<sup>−1</sup>.</p>
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<p>Difference in survivorship of red porgy larvae exposed to different concentrations of <span class="html-italic">K. brevis</span> strain NCMA2228 for either 24 or 48 h. (<b>A</b>) Exposure of four days post hatch larvae, (<b>B</b>) six days post hatch, and (<b>C</b>) sixteen days post hatch larvae.</p>
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15 pages, 4865 KiB  
Article
Shining Light on Photosynthesis in the Harmful Dinoflagellate Karenia mikimotoi–Responses to Short-Term Changes in Temperature, Nitrogen Form, and Availability
by So Hyun (Sophia) Ahn and Patricia M. Glibert
Phycology 2022, 2(1), 30-44; https://doi.org/10.3390/phycology2010002 - 27 Dec 2021
Cited by 4 | Viewed by 3212
Abstract
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 [...] Read more.
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
(This article belongs to the Collection Feature Papers in Phycology)
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<p>Schematic of the experimental design to measure <sup>15</sup>N incorporation and photosynthesis-irradiance responses of <span class="html-italic">K. mikimotoi</span> exposed to different temperatures, N forms, and concentrations. Green tubes represent those enriched with urea; blue tubes—those enriched with NH<sub>4</sub><sup>+</sup>; and red tubes—those enriched with NO<sub>3</sub><sup>−</sup>.</p>
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<p>Maximum photosystem II photochemical efficiency, Fv/Fm, of <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM-N). Within each depicted temperature, the darker the color of the bars, the higher the enrichment with added N.</p>
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<p>Effective Photosystem II photochemical efficiency, YII, as a function of irradiance for <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM-N). Within each depicted irradiance level, the darker the color of the bars, the higher the enrichment with added N.</p>
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<p>The rapid light responses of rETR of <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM-N). For each depicted ETR curve, the darker the color of the lines, the higher the enrichment with added N.</p>
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<p>The rETR<sub>max</sub> of <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM). Within each depicted temperature, the darker the color of the bars, the higher the enrichment with added N.</p>
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<p>The extent of PSII reaction center closures, 1-qP, as a function of irradiance for <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM-N). Within each depicted irradiance level, the darker the color of the bars, the higher the enrichment with added N.</p>
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<p>Atom percent enrichment of <sup>15</sup>N for <span class="html-italic">K. mikimotoi</span> exposed to different temperatures (15, 20, 25, 30 °C) when pulsed with different nitrogen forms (NO<sub>3</sub><sup>−</sup>, NH<sub>4</sub><sup>+</sup>, urea) and amounts (1, 5, 10, 20, 50 µM-N). For each depicted curve, the darker the color of the lines, the higher the enrichment with added N. Note that the scale of Y-axis is different.</p>
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<p>Conceptual figure representing push and pull of energy during photosynthesis and metabolic reactions associated with C and N assimilation.</p>
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11 pages, 1662 KiB  
Article
Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay
by Xiya Zhang, Mingyue Ding, Chensi Zhang, Yexuan Mao, Youyi Wang, Peipei Li, Haiyang Jiang, Zhanhui Wang and Xuezhi Yu
Foods 2021, 10(10), 2398; https://doi.org/10.3390/foods10102398 - 9 Oct 2021
Cited by 10 | Viewed by 2416
Abstract
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 [...] Read more.
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 ?g/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 ?g/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
(This article belongs to the Special Issue Application of Antibody and Immunoassay for Food Safety)
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<p>PbTx-1, -2, and -3 structures.</p>
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<p>PbTx-2 hapten and antigen synthesis routes. (<b>A</b>) PbTx-2-CMO-KLH/BSA. (<b>B</b>) PbTx-2HS-KLH/BSA.</p>
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<p>Molecular modeling results. (<b>A</b>) Overlap of PbTx-2 (gray), PbTx-2-CMO (green), and PbTx-2-HS (violet) structures. (<b>B</b>) Calculated partial atomic charges of PbTx-2, PbTx-2-CMO, and PbTx-2-HS structures. (<b>C</b>) ESP for PbTx-2, PbTx-2-CMO, and PbTx-2-HS structures. Red and blue areas indicate negative and positive potentials, respectively.</p>
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<p>UV–visible absorption spectra of PbTx-2-CMO-KLH (<b>A</b>), PbTx-2-CMO-BSA (<b>B</b>), PbTx-2-CMO-KLH (<b>C</b>) and PbTx-2-CMO-BSA (<b>D</b>).</p>
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<p>The brevetoxin icELISA standard curve using the mAb 1D3 with the PbTx-2-HS-BSA coating antigen.</p>
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22 pages, 2663 KiB  
Article
A Remote Sensing and Machine Learning-Based Approach to Forecast the Onset of Harmful Algal Bloom
by Moein Izadi, Mohamed Sultan, Racha El Kadiri, Amin Ghannadi and Karem Abdelmohsen
Remote Sens. 2021, 13(19), 3863; https://doi.org/10.3390/rs13193863 - 27 Sep 2021
Cited by 42 | Viewed by 7264
Abstract
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, [...] Read more.
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
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<p>Location map for the study area covering coastal waters (width: ~10–60 km; length: 180 km) of Charlotte County in southwest Florida.</p>
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<p>Flowchart describing the adopted methodology.</p>
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<p>Two-dimensional feature space with SVM linear discrimination function.</p>
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<p>Machine Learning models ROCs for Test ROC (<b>a</b>) and Train ROC (<b>b</b>).</p>
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<p>Comparison between the reported <span class="html-italic">K. brevis</span> concentration in 170 randomly selected test samples within the study area (<b>top</b>) with their predicted concentrations from an 8-day RF model (<b>bottom</b>). The samples have been classified correctly with 95% accuracy.</p>
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<p>Variable importance (VI) boxplot for the best RF model. The VI metrics are on the y-axis and the 10 variables for each of the 3 consecutive days are numbered.</p>
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14 pages, 6266 KiB  
Article
Guidance Level for Brevetoxins in French Shellfish
by Nathalie Arnich, Eric Abadie, Zouher Amzil, Marie-Yasmine Dechraoui Bottein, Katia Comte, Estelle Chaix, Nicolas Delcourt, Vincent Hort, César Mattei, Jordi Molgó and Raphaele Le Garrec
Mar. Drugs 2021, 19(9), 520; https://doi.org/10.3390/md19090520 - 15 Sep 2021
Cited by 19 | Viewed by 4045
Abstract
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 [...] Read more.
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. Full article
(This article belongs to the Special Issue Marine Biotoxins)
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<p>Location of the Diana lagoon in Corsica Island, Mediterranean Sea (red bookmark, © OpenStreetMap contributors. Tile style for the Humanitarian OpenStreetMap Team hosted by OpenStreetMap France). (<b>A</b>) Location of Corsica Island in Europe. (<b>B</b>) Location of Diana lagoon in Corsica Island. (<b>C</b>) Enlargement of the area of the Diana lagoon.</p>
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