Influence of multi-stressor combinations of pCO2, temperature, and salinity on the toxicity of Heterosigma akashiwo (Raphidophyceae), a fish-killing flagellate.

Climate change and global warming have led to more frequent harmful algal blooms in the last decade. Among these blooms, Heterosigma akashiwo, a golden-brown phytoflagellate, is one of the 40 species with a high potential to form harmful blooms, leading to significant fish mortality. Climate change leads to rising atmospheric and ocean temperatures. These changes, along with altered rainfall patterns and meltwater input, can cause fluctuations in ocean salinity. Elevated atmospheric carbon dioxide (CO2) levels increase water acidity as oceans absorb CO2. This study investigated the effects of temperature, salinity, and CO2 levels on lipid production, hemolytic activity, and toxicity of H. akashiwo using the design of experiment approach, which can be used to investigate the effect of two or more factors on the same response simultaneously in a precise manner with fewer experiments and materials but in a larger region of the factor space. The lipid content was measured using a high-throughput Nile Red method, and the highest level of lipid content was detected at 25°C, a salinity of 30, and a CO2 concentration of 400 ppm. Hemolytic activity was assessed using rabbit blood erythrocytes in a 96-well plate, and the optimal conditions for achieving the highest hemolytic activity were determined at 15°C, a salinity of 10, and a CO2 concentration of 400 ppm. As the chemical structure of the toxin is not known, we used the toxicity against the cell line RTgill-W1 as the cell toxicity proxy. The maximum toxicity was identified at 15°C, a salinity of 10, and a CO2 level of 700 ppm.

Climate change amplifies the risk of potentially toxigenic cyanobacteria.

Cyanobacterial blooms pose a significant threat to water security, with anthropogenic forcing being implicated as a key driver behind the recent upsurge and global expansion of cyanobacteria in modern times. The potential effects of land-use alterations and climate change can lead to complicated, less-predictable scenarios in cyanobacterial management, especially when forecasting cyanobacterial toxin risks. There is a growing need for further investigations into the specific stressors that stimulate cyanobacterial toxins, as well as resolving the uncertainty surrounding the historical or contemporary nature of cyanobacterial-associated risks. To address this gap, we employed a paleolimnological approach to reconstruct cyanobacterial abundance and microcystin-producing potential in temperate lakes situated along a human impact gradient. We identified breakpoints (i.e., points of abrupt change) in these time series and examined the impact of landscape and climatic properties on their occurrence. Our findings indicate that lakes subject to greater human influence exhibited an earlier onset of cyanobacterial biomass by 40 years compared to less-impacted lakes, with land-use change emerging as the dominant predictor. Moreover, microcystin-producing potential increased in both high- and low-impact lakes around the 1980s, with climate warming being the primary driver. Our findings chronicle the importance of climate change in increasing the risk of toxigenic cyanobacteria in freshwater resources.

Insights into Cellular Localization and Environmental Influences on the Toxicity of Marine Fish-Killing Flagellate, Heterosigma akashiwo.

Heterosigma akashiwo is a unicellular microalga which can cause massive mortality in both wild and cultivated fish worldwide, resulting in substantial economic losses. Environmental parameters such as salinity, light, and temperature showed a significant effect on bloom initiation and the toxicity of H. akashiwo. While in previous studies a one-factor-at-a-time (OFAT) approach was utilized, which only changes one variable at a time while keeping others constant, in the current study a more precise and effective design of experiment (DOE) approach, was used to investigate the simultaneous effect of three factors and their interactions. The study employed a central composite design (CCD) to investigate the effect of salinity, light intensity, and temperature on the toxicity, lipid, and protein production of H. akashiwo. A yeast cell assay was developed to assess toxicity, which offers rapid and convenient cytotoxicity measurements using a lower volume of samples compared to conventional methods using the whole organism. The obtained results showed that the optimum condition for toxicity of H. akashiwo was 25 °C, a salinity of 17.5, and a light intensity of 250 µmol photons m-2 s-1. The highest amount of lipid and protein was found at 25 °C, a salinity of 30, and a light intensity of 250 µmol photons m-2 s-1. Consequently, the combination of warm water mixing with lower salinity river input has the potential to enhance H. akashiwo toxicity, which aligns with environmental reports that establish a correlation between warm summers and extensive runoff conditions that indicate the greatest concern for aquaculture facilities.

Reconstructing historical time-series of cyanobacteria in lake sediments: Integrating technological innovation to enhance cyanobacterial management.

The global rise of cyanobacterial blooms emphasizes the need to develop tools to manage water bodies prone to cyanobacterial dominance. Reconstructing cyanobacterial baselines and identifying environmental drivers that favour cyanobacterial dominance are important to guide management decisions. Conventional techniques for estimating cyanobacteria in lake sediments require considerable resources, creating a barrier to routine reconstructions of cyanobacterial time-series. Here, we compare a relatively simple technique based on spectral inferences of cyanobacteria using visible near-infrared reflectance spectroscopy (VNIRS) with a molecular technique based on real-time PCR quantification (qPCR) of the 16S rRNA gene conserved in cyanobacteria in 30 lakes across a broad geographic gradient. We examined the sedimentary record from two perspectives: 1) relationships throughout the entire core (without radiometric dating); 2) relationships post-1900s with the aid of radiometric dating (i.e., 210Pb). Our findings suggest that the VNIRS-based cyanobacteria technique is best suited for reconstructing cyanobacterial abundance in recent decades (i.e., circa 1990 onwards). The VNIRS-based cyanobacteria technique showed agreement with those generated using qPCR, with 23 (76%) lakes showing a strong or very strong positive relationship between the results of the two techniques. However, five (17%) lakes showed negligible relationships, suggesting cyanobacteria VNIRS requires further refinement to understand where VNIRS is unsuitable. This knowledge will help scientists and lake managers select alternative cyanobacterial diagnostics where appropriate. These findings demonstrate the utility of VNIRS, in most instances, as a valuable tool for reconstructing past cyanobacterial prevalence.

Deep Cyanobacteria Layers: An Overlooked Aspect of Managing Risks of Cyanobacteria.

The risk of human exposure to cyanotoxins is partially influenced by the location of toxin-producing cyanobacteria in waterbodies. Cyanotoxin production can occur throughout the water column, with deep water production representing a potential public health concern, specifically for drinking water supplies. Deep cyanobacteria layers are often unreported, and it remains to be seen if lower incident rates reflect an uncommon phenomenon or a monitoring bias. Here, we examine Sunfish Lake, Ontario, Canada as a case study lake with a known deep cyanobacteria layer. Cyanotoxin and other bioactive metabolite screening revealed that the deep cyanobacteria layer was toxigenic [0.03 µg L-1 microcystins (max) and 2.5 µg L-1 anabaenopeptins (max)]. The deep layer was predominantly composed of Planktothrix isothrix (exhibiting a lower cyanotoxin cell quota), with Planktothrix rubescens (exhibiting a higher cyanotoxin cell quota) found at background levels. The co-occurrence of multiple toxigenic Planktothrix species underscores the importance of routine surveillance for prompt identification leading to early intervention. For instance, microcystin concentrations in Sunfish Lake are currently below national drinking water thresholds, but shifting environmental conditions (e.g., in response to climate change or nutrient modification) could fashion an environment favoring P. rubescens, creating a scenario of greater cyanotoxin production. Future work should monitor the entire water column to help build predictive capacities for identifying waterbodies at elevated risk of developing deep cyanobacteria layers to safeguard drinking water supplies.

Differential Drawdown of Ammonium, Nitrate, and Urea by Freshwater Chlorophytes and Cyanobacteria1.

The chemical form of nitrogen (N) is deemed to be decisive in shaping the composition of the primary producer community. Recently, there has been a shift in the dominant form of N delivered to agricultural landscapes. Urea-based fertilizers are a mainstay in modern agriculture, and their ubiquitous use has increased the likelihood of urea export to nearby freshwaters. The shift to urea fertilizers has coincided with the recent expansion of cyanobacteria harmful algal blooms (cyanoHABs). This study investigated N drawdown patterns between two major freshwater phytoplankton groups-chlorophytes and cyanobacteria. Experiments were designed to understand if different patterns of N drawdown occurred among taxa and the potential synergistic effects of multiple N substrates. Nitrate (NO3- ), ammonium (NH4+ ), and urea were supplied in a series of paired combinations, and N concentrations were monitored to track N drawdowns. We did not find significant differences between phytoplankton classes when supplied with a single N substrate. However, we found that when N substrates were supplied in combination, significant differences in N drawdown patterns were observed. Urea was consumed more rapidly among cyanobacteria, being drawn down at significantly higher rates relative to inorganic N substrates. In contrast, inorganic N substrates were drawn down more rapidly among chlorophytes relative to urea. Our findings support the emerging urea-cyanoHAB link and the potential importance of urea in freshwater eutrophication. As society becomes increasingly dependent on urea for agricultural crops, the need to understand how urea influences phytoplankton community composition may be instrumental in predicting bloom dynamics.

Global change-driven effects on dissolved organic matter composition: Implications for food webs of northern lakes.

Northern ecosystems are experiencing some of the most dramatic impacts of global change on Earth. Rising temperatures, hydrological intensification, changes in atmospheric acid deposition and associated acidification recovery, and changes in vegetative cover are resulting in fundamental changes in terrestrial-aquatic biogeochemical linkages. The effects of global change are readily observed in alterations in the supply of dissolved organic matter (DOM)-the messenger between terrestrial and lake ecosystems-with potentially profound effects on the structure and function of lakes. Northern terrestrial ecosystems contain substantial stores of organic matter and filter or funnel DOM, affecting the timing and magnitude of DOM delivery to surface waters. This terrestrial DOM is processed in streams, rivers, and lakes, ultimately shifting its composition, stoichiometry, and bioavailability. Here, we explore the potential consequences of these global change-driven effects for lake food webs at northern latitudes. Notably, we provide evidence that increased allochthonous DOM supply to lakes is overwhelming increased autochthonous DOM supply that potentially results from earlier ice-out and a longer growing season. Furthermore, we assess the potential implications of this shift for the nutritional quality of autotrophs in terms of their stoichiometry, fatty acid composition, toxin production, and methylmercury concentration, and therefore, contaminant transfer through the food web. We conclude that global change in northern regions leads not only to reduced primary productivity but also to nutritionally poorer lake food webs, with discernible consequences for the trophic web to fish and humans.

The effects of salinity on the cellular permeability and cytotoxicity of Heterosigma akashiwo.

A laboratory study using the fish-killing raphidophyte Heterosigma akashiwo was conducted to examine its capability to grow at salinities below oceanic, and to test the perceived relationship between reduced salinities and increased cytotoxicity. A nonaxenic strain of H. akashiwo isolated from the U.S. Pacific Northwest was exposed to a combination of three salinity (32, 20, and 10) and five temperature (14.7°C, 18.4°C, 21.4°C, 24.4°C and 27.8°C) conditions. Our results demonstrate that cell permeability and cytotoxicity are strongly correlated in unialgal cultures of H. akashiwo, which both increased as salinity decreased from 32 to 10. Furthermore, over a broad median range of salinities (10 and 20), neither temperature nor specific growth rate was correlated with cytotoxicity. However, in cultures grown at the salinity of 32, both temperature and specific growth rate were inversely proportional to toxicity; this relationship was likely due to the effect of contamination by an unidentified species of Skeletonema in those cultures. The presence of Skeletonema sp. resulted in a cytotoxic response from H. akashiwo that was greater than the response caused by salinity alone. These laboratory results reveal the capability of H. akashiwo to become more toxic not only at reduced salinities but also in competition with another algal species. Changes in cell permeability in response to salinity may be an acclimation mechanism by which H. akashiwo is able to respond rapidly to different salinities. Furthermore, due to its strong positive correlation with cytotoxicity, cellular permeability is potentially associated with the ichthyotoxic pathway of this raphytophyte.

Harmful algal blooms and climate change: Learning from the past and present to forecast the future.

Climate change pressures will influence marine planktonic systems globally, and it is conceivable that harmful algal blooms may increase in frequency and severity. These pressures will be manifest as alterations in temperature, stratification, light, ocean acidification, precipitation-induced nutrient inputs, and grazing, but absence of fundamental knowledge of the mechanisms driving harmful algal blooms frustrates most hope of forecasting their future prevalence. Summarized here is the consensus of a recent workshop held to address what currently is known and not known about the environmental conditions that favor initiation and maintenance of harmful algal blooms. There is expectation that harmful algal bloom (HAB) geographical domains should expand in some cases, as will seasonal windows of opportunity for harmful algal blooms at higher latitudes. Nonetheless there is only basic information to speculate upon which regions or habitats HAB species may be the most resilient or susceptible. Moreover, current research strategies are not well suited to inform these fundamental linkages. There is a critical absence of tenable hypotheses for how climate pressures mechanistically affect HAB species, and the lack of uniform experimental protocols limits the quantitative cross-investigation comparisons essential to advancement. A HAB "best practices" manual would help foster more uniform research strategies and protocols, and selection of a small target list of model HAB species or isolates for study would greatly promote the accumulation of knowledge. Despite the need to focus on keystone species, more studies need to address strain variability within species, their responses under multifactorial conditions, and the retrospective analyses of long-term plankton and cyst core data; research topics that are departures from the norm. Examples of some fundamental unknowns include how larger and more frequent extreme weather events may break down natural biogeographic barriers, how stratification may enhance or diminish HAB events, how trace nutrients (metals, vitamins) influence cell toxicity, and how grazing pressures may leverage, or mitigate HAB development. There is an absence of high quality time-series data in most regions currently experiencing HAB outbreaks, and little if any data from regions expected to develop HAB events in the future. A subset of observer sites is recommended to help develop stronger linkages among global, national, and regional climate change and HAB observation programs, providing fundamental datasets for investigating global changes in the prevalence of harmful algal blooms. Forecasting changes in HAB patterns over the next few decades will depend critically upon considering harmful algal blooms within the competitive context of plankton communities, and linking these insights to ecosystem, oceanographic and climate models. From a broader perspective, the nexus of HAB science and the social sciences of harmful algal blooms is inadequate and prevents quantitative assessment of impacts of future HAB changes on human well-being. These and other fundamental changes in HAB research will be necessary if HAB science is to obtain compelling evidence that climate change has caused alterations in HAB distributions, prevalence or character, and to develop the theoretical, experimental, and empirical evidence explaining the mechanisms underpinning these ecological shifts.

Cultural and socio-economic conditions as factors contributing to chronic stress in sub-Saharan African communities.

Stress is known to contribute to overall health status. Many individuals in sub-Saharan Africa are believed to be stressed about their employment, income, and health. This study aimed to investigate hair cortisol as a biomarker of chronic stress in settlement communities in Kenya. Hair samples were collected from 108 volunteers from settlement communities in Kenya. An enzyme-linked immunosorbent assay technique was used to measure hair cortisol concentrations. In parallel, a health survey was completed. The mean ± SD for the cortisol concentration in the hair of volunteers from the settlement communities in Naivasha was 639 ± 300 ng/g, which was higher than found for a Caucasian reference group (299 ± 110 ng/g; one-way ANOVA, P = 0.0003). There were no differences in hair cortisol concentrations between members of slum settlements adjacent to large floriculture farms in Naivasha (Karagita, Kamere/Kwa Muhia/DCK, and Kasarani) compared with those well-removed from all floriculture in Mogotio (Mogotio and Westlands/Katorongot). However, hair cortisol concentrations were significantly higher in females, divorced volunteers, those who made below minimum wage, and those who reported feeling unsafe collecting water or using sanitation facilities within these 2 settlement groups. We found no evidence for increased chronic stress (measured by hair cortisol content) between members of slum settlements adjacent to versus distant to large floriculture farms. Cultural and socio-economic conditions that prevail in much of sub-Saharan Africa were found to be factors contributing to chronic stress.

A profile of an endosymbiont-enriched fraction of the coral Stylophora pistillata reveals proteins relevant to microbial-host interactions.

This study examines the response of Symbiodinium sp. endosymbionts from the coral Stylophora pistillata to moderate levels of thermal "bleaching" stress, with and without trace metal limitation. Using quantitative high throughput proteomics, we identified 8098 MS/MS events relating to individual peptides from the endosymbiont-enriched fraction, including 109 peptides meeting stringent criteria for quantification, of which only 26 showed significant change in our experimental treatments; 12 of 26 increased expression in response to thermal stress with little difference affected by iron limitation. Surprisingly, there were no significant increases in antioxidant or heat stress proteins; those induced to higher expression were generally involved in protein biosynthesis. An outstanding exception was a massive 114-fold increase of a viral replication protein indicating that thermal stress may substantially increase viral load and thereby contribute to the etiology of coral bleaching and disease. In the absence of a sequenced genome for Symbiodinium or other photosymbiotic dinoflagellate, this proteome reveals a plethora of proteins potentially involved in microbial-host interactions. This includes photosystem proteins, DNA repair enzymes, antioxidant enzymes, metabolic redox enzymes, heat shock proteins, globin hemoproteins, proteins of nitrogen metabolism, and a wide range of viral proteins associated with these endosymbiont-enriched samples. Also present were 21 unusual peptide/protein toxins thought to originate from either microbial consorts or from contamination by coral nematocysts. Of particular interest are the proteins of apoptosis, vesicular transport, and endo/exocytosis, which are discussed in context of the cellular processes of coral bleaching. Notably, the protein complement provides evidence that, rather than being expelled by the host, stressed endosymbionts may mediate their own departure.

A geo-gender-based analysis of human health: The presence of cut flower farms can attenuate pesticide exposure in African communities, with women being the most vulnerable.

Background: The rapid expansion of the cut flower industry in Africa has led to pervasive use and potential exposure of pesticides, raising concerns for local communities. Whether the risks associated with pesticide applications are localised or have broader implications remains unclear. Methods: We measured biomarkers of real and perceived pesticide exposure in two Kenyan communities: Naivasha, where the cut flower industry is present, and Mogotio, where the cut flower industry is absent. We measured real exposure by the percentage of acetylcholinesterase (AChE) inhibition and perceived exposure by assessing hair cortisol levels, a biomarker of stress. Additionally, we conducted a demographic survey to evaluate the health and socioeconomic status of participants, as well as their perceptions of pesticide risks associated with the cut flower industry. Results: Perceived pesticide exposure was more common in Naivasha (n = 36, 56%) compared to Mogotio (n = 0, 0%), according to community surveys. However, Mogotio residents had significantly higher mean hair cortisol levels (mean (x̄) = 790 ng/g, standard deviation (SD) = 233) and percentage of AChE inhibition (x̄ = 28.5%, SD = 7.3) compared to Naivasha residents, who had lower mean hair cortisol levels (x̄ = 548 ng/g, SD = 187) and percentage of AChE inhibition (x̄ = 14.5%, SD = 10.1). Location (proximity to cut flower farms) and gender were significant factors influencing pesticide exposure, with individuals living outside the cut flower industrial complexes being at higher risk. Women in both communities were the most vulnerable demographic, showing significantly higher mean hair cortisol levels (x̄ = 646 ng/g, SD = 267.4) and percentage of AChE inhibition (x̄ = 22.5%, SD = 12.4) compared to men hair cortisol levels (x̄ = 558.2 ng/g, SD = 208.2) and percentage of AChE inhibition (x̄ = 10.4%, SD = 13.1). Conclusions: A heightened awareness of the potential risks of pesticide exposure was widespread within cut flower industrial complexes. This may have led to a reduction in exposure of both workers and non-workers living within or close to these complexes. In contrast, communities living outside these complexes showed higher levels of exposure, possibly due to limited chemical awareness and a lack of precautionary measures. Despite this contrast between communities, women remained the most vulnerable members, likely due to their socioeconomic roles in African society. Monitoring women's pesticide exposure is crucial for providing an early warning system for community exposure.

Iron enrichment stimulates toxic diatom production in high-nitrate, low-chlorophyll areas.

Oceanic high-nitrate, low-chlorophyll environments have been highlighted for potential large-scale iron fertilizations to help mitigate global climate change. Controversy surrounds these initiatives, both in the degree of carbon removal and magnitude of ecosystem impacts. Previous open ocean enrichment experiments have shown that iron additions stimulate growth of the toxigenic diatom genus Pseudonitzschia. Most Pseudonitzschia species in coastal waters produce the neurotoxin domoic acid (DA), with their blooms causing detrimental marine ecosystem impacts, but oceanic Pseudonitzschia species are considered nontoxic. Here we demonstrate that the sparse oceanic Pseudonitzschia community at the high-nitrate, low-chlorophyll Ocean Station PAPA (50 degrees N, 145 degrees W) produces approximately 200 pg DA L(-1) in response to iron addition, that DA alters phytoplankton community structure to benefit Pseudonitzschia, and that oceanic cell isolates are toxic. Given the negative effects of DA in coastal food webs, these findings raise serious concern over the net benefit and sustainability of large-scale iron fertilizations.

Harmonizing science and management options to reduce risks of cyanobacteria.

Management of cyanobacteria has become an increasingly complex venture. Cyanobacteria risks have amplified as society moves forward in an era of accelerated global changes. The cyanobacteria management "pendulum" has progressively shifted from prevention to mitigation, with management considerations often put forth after bloom formation. A universal system (i.e., one-size-fits-all management) fails to provide a management path forward due to the inherent complexities of each lake. A tailored management plan is needed: the right species at the right time in the right place (i.e., the three Rs). The three Rs represent a customizable management strategy that is flexible and informed by advances in scientific understanding to lower cyanobacteria-associated risks. Identifying thresholds in risk tolerance, where thresholds are defined by community collectives, is essential to frame cyanobacteria management targets and to decide on what management interventions are warranted.

Palmelloid formation in the Antarctic psychrophile, Chlamydomonas priscuii, is photoprotective.

Cultures of the obligate, Antarctic psychrophile, Chlamydomonas priscuii grown at permissive low temperature (8°C) are composed of flagellated, single cells, as well as non-motile, multicellular palmelloids. The relative proportions of the two cell types are temperature dependent. However, the temperature dependence for palmelloid formation is not restricted to psychrophilic C. priscuii but appears to be a general response of mesophilic Chlamydomonas species (C. reinhardtii and C. raudensis) to non-permissive growth temperatures. To examine potential differences in photosynthetic performance between single cells versus palmelloids of the psychrophile, a cell filtration technique was developed to separate single cells from palmelloids of C. priscuii grown at 8°C. Flow cytometry was used to estimate the diameter of isolated single cells (≤5 µm) versus isolated palmelloids of varying size (≥8 µm). Compared to single cells, palmelloids of C. priscuii showed a decrease in the abundance of light-harvesting complex II (LHCII) proteins with a 2-fold higher Chl a/b ratio. A decrease in both lutein and ß-carotene in palmelloids resulted in carotenoid pools which were 27% lower in palmelloids compared to single cells of the psychrophile. Chlorophyll fluorescence analyses of the isolated fractions revealed that maximum photochemical efficiency of PSII (Fv/Fm) was comparable for both single cells and palmelloids of C. priscuii. However, isolated palmelloids exhibited lower excitation pressure, measured as 1 - qL, but higher yield of PSII (ΦPSII) and 50% higher rates of electron transport (ETR) than single cells exposed to high light at 8°C. This decreased sensitivity to high light in isolated palmelloids compared to single cells was associated with greater non-regulated dissipation of excess absorbed energy (ΦNO) with minimal differences in ΦNPQ in C. priscuii in response to increasing irradiance at low temperature. The ratio ΦNO/ΦNPQ observed for isolated palmelloids of C. priscuii developed at 8°C (1.414 ± 0.036) was 1.38-fold higher than ΦNO/ΦNPQ of isolated single cells (1.021 ± 0.018) exposed to low temperature combined with high light (1,000 µmol m-2 s-1). The differences in the energy quenching capacities between palmelloids and single cells are discussed in terms of enhanced photoprotection of C. priscuii palmelloids against low-temperature photoinhibition.

The growing need for sustainable ecological management of marine communities of the Persian Gulf.

The Persian Gulf is a semi-enclosed marine system surrounded by eight countries, many of which are experiencing substantial development. It is also a major center for the oil industry. The increasing array of anthropogenic disturbances may have substantial negative impacts on marine ecosystems, but this has received little attention until recently. We review the available literature on the Gulfs marine environment and detail our recent experience in the United Arab Emirates (U.A.E.) to evaluate the role of anthropogenic disturbance in this marine ecosystem. Extensive coastal development may now be the single most important anthropogenic stressor. We offer suggestions for how to build awareness of environmental risks of current practices, enhance regional capacity for coastal management, and build cooperative management of this important, shared marine system. An excellent opportunity exists for one or more of the bordering countries to initiate a bold and effective, long-term, international collaboration in environmental management for the Gulf.

Yeast Cell as a Bio-Model for Measuring the Toxicity of Fish-Killing Flagellates.

Harmful algal blooms are a significant environmental problem. Cells that bloom are often associated with intercellular or dissolved toxins that are a grave concern to humans. However, cells may also excrete compounds that are beneficial to their competition, allowing the cells to establish or maintain cells in bloom conditions. Here, we develop a yeast cell assay to assess whether the bloom-forming species can change the toxicity of the water environment. The current methods of assessing toxicity involve whole organisms. Here, yeast cells are used as a bioassay model to evaluate eukaryotic cell toxicity. Yeast is a commonly used, easy to maintain bioassay species that is free from ethical concerns, yet is sensitive to a wide array of metabolic and membrane-modulating agents. Compared to methods in which the whole organism is used, this method offers rapid and convenient cytotoxicity measurements using a lower volume of samples. The flow cytometer was employed in this toxicology assessment to measure the number of dead cells using alive/dead stain analysis. The results show that yeast cells were metabolically damaged after 1 h of exposure to our model toxin-producing euryhaline flagellates (Heterosigma akashiwo and Prymnesium parvum) cells or extracts. This amount was increased by extending the incubation time.

Insensitivities of a subtropical productive coastal plankton community and trophic transfer to ocean acidification: Results from a microcosm study.

Ocean acidification (OA) has potential to affect marine phytoplankton in ways that are partly understood, but there is less knowledge about how it may alter the coupling to secondary producers. We investigated the effects of OA on phytoplankton primary production, and its trophic transfer to zooplankton in a subtropical eutrophic water (Wuyuan Bay, China) under present day (400 µatm) and projected end-of-century (1000 µatm) pCO2 levels. Net primary production was unaffected, although OA did lead to small decreases in growth rates. OA had no measurable effect on micro-/mesozooplankton grazing rates. Elevated pCO2 had no effect on phytoplankton fatty acid (FA) concentrations during exponential phase, but saturated FAs increased relative to the control during declining phase. FA profiles of mesozooplankton were unaffected. Our findings show that short-term exposure of plankton communities in eutrophic subtropical waters to projected end-of-century OA conditions has little effect on primary productivity and trophic linkage to mesozooplankton.

The successional formation and release of domoic acid in a Pseudo-nitzschia bloom in the Juan de Fuca Eddy: A drifter study.

Blooms of Pseudo-nitzschia species are frequent, but presently unpredictable, in the Juan de Fuca Eddy region off the coasts of Washington (US) and British Columbia (Canada). This upwelling eddy region is proposed to be the bloom commencement site, before cells are entrained into the coastal surface currents. During a shipboard study, we characterized the different stages of the Pseudo-nitzschia bloom development from its initiation and intensification, to its eventual sinking and dissipation. Specifically, we followed a water mass using lagrangian ARGOS-tracked drifters released at the eddy water mass and quantified production of dissolved and particulate domoic acid, and the physiological status of the Pseudo-nitzschia cells with regards to photosynthesis, nutrient needs and sinking rates, along with its relationship with competing species - in this case, the marine euglenoid, Eutreptiella spp. The drifter study allows for an interpretation of the presence or absence of Pseudo-nitzschia and domoic acid against active environmental factors - particularly copper and iron.

Suitability of a cytotoxicity assay for detection of potentially harmful compounds produced by freshwater bloom-forming algae.

Detecting harmful bioactive compounds produced by bloom-forming pelagic algae is important to assess potential risks to public health. We investigated the application of a cell-based bioassay: the rainbow trout gill-w1 cytotoxicity assay (RCA) that detects changes in cell metabolism. The RCA was used to evaluate the cytotoxic effects of (1) six natural freshwater lake samples from cyanobacteria-rich lakes in central Ontario, Canada; (2) analytical standards of toxins and noxious compounds likely to be produced by the algal communities in these lakes; and (3) complex mixtures of compounds produced by cyanobacterial and chrysophyte cultures. RCA provided a measure of lake water toxicity that could not be reproduced using toxin or noxious compound standards. RCA was not sensitive to toxins and only sensitive to noxious compounds at concentrations higher than reported environmental averages (EC50≥103nM). Cultured algae produced bioactive compounds that had recognizable dose dependent and toxic effects as indicated by RCA. Toxicity of these bioactive compounds depended on taxa (cyanobacteria, not chrysophytes), growth stage (stationary phase more toxic than exponential phase), location (intracellular more toxic than extracellular) and iron status (cells in high-iron treatment more toxic than cells in low-iron treatment). The RCA provides a new avenue of exploration and potential for the detection of natural lake algal toxic and noxious compounds.