Physiological responses of the microalga Isochrysis galbana exposed to polystyrene microplastics with different particle sizes.

Due to continuous increase in marine plastic waste, microplastics are ubiquitous in the marine environment. However, there are few studies on the harmful effects caused by microplastics with different particle sizes, and the interaction between particle size and concentration requires further investigation. This study explored the differences in physiological and biochemical responses, photosynthesis and oxidative stress damage of the microalga Isochrysis galbana exposed to three different particle size microplastics. It was found that different particle sizes and concentrations of microplastics resulted in significant differences (p < 0.05) in the growth rate, photosynthesis, and oxidative stress level of I. galbana. With the decrease of the particle size and lowering concentration of microplastics, the growth rate, photosynthesis and oxidative stress levels of I. galbana were reduced. Significant differences in photosynthesis and oxidative stress levels were observed when I. galbana was exposed to smallest particle size and lowest concentration of microplastics. This study provides new insights about whether polystyrene microplastics of different particle sizes and concentrations exhibit complex effects on microalgae, and explores the underlying reasons for such effects. In short, this study predicts the exacerbating adverse effects of microplastic pollution on the primary productivity, with significant implications for marine food webs and ecosystem health.

Unpredicted ecosystem response to compound human impacts in a European river.

Climate change elevates the threat of compound heat and drought events, with their ecological and socioeconomic impacts exacerbated by human ecosystem alterations such as eutrophication, salinization, and river engineering. Here, we study how multiple stressors produced an environmental disaster in a large European river, the Oder River, where a toxic bloom of the brackish-water planktonic haptophyte Prymnesium parvum (the "golden algae") killed approximately 1000 metric tons of fish and most mussels and snails. We uncovered the complexity of this event using hydroclimatic data, remote sensing, cell counts, hydrochemical and toxin analyses, and genetics. After incubation in impounded upstream channels with drastically elevated concentrations of salts and nutrients, only a critical combination of chronic salt and nutrient pollution, acute high water temperatures, and low river discharge during a heatwave enabled the riverine mass proliferation of B-type P. parvum along a 500 km river section. The dramatic losses of large filter feeders and the spreading of vegetative cells and resting stages make the system more susceptible to new harmful algal blooms. Our findings show that global warming, water use intensification, and chronic ecosystem pollution could increase likelihood and severity of such compound ecoclimatic events, necessitating consideration in future impact models.

A rapid and multi-endpoint ecotoxicological test using Mychonastes afer for efficient screening of metals and herbicides.

Microalgal growth-based tests are international standards for ecotoxicity assessment; however, their long exposure times, large sample volumes, and reliance on a single growth-endpoint make them inadequate for rapid toxicity screening. Here, we aimed to develop a rapid and simple ecotoxicological test using the fast-growing green alga Mychonastes afer, with multiple endpoints-growth, lipid content, and photosynthesis. We exposed M. afer to two metals-silver and copper-and two herbicides-atrazine and diuron-for 24 h and identified the most sensitive and reliable endpoints for each toxicant: the maximum electron transport rate (ETRmax) for Ag, Cu and atrazine, and the lipid content for diuron. Lipid content was found to be both a sensitive and reliable biomarker, meeting the effluent limit guidelines in both the Republic of Korea and the USA. The sensitivity of M. afer to Ag and atrazine also closely matched the HC5 values derived from the species sensitivity distribution approach, confirming its reliability for setting regulatory concentrations of these contaminants. Our calculated predicted no-effect concentration (PNEC) values were similar to established European Union PNECs for Ag, Cu, atrazine, and diuron, underlining the utility of these biological endpoints for ecological risk assessment and regulatory decision making. This method required lower sample volume (2 mL vs 100 mL) and exposure time (24 h vs 72-120 h) than conventional green algal tests, and eliminated the need for labour-intensive cell counting, expensive equipment, and chlorophyll fluorescence measurement expertise. Overall, this M. afer test can be a valuable tool for the rapid screening of wastewater for metals and herbicides, contributing to environmental protection and management practices.

Effects of micro- and nano-plastics on growth, antioxidant system, DMS, and DMSP production in Emiliania huxleyi.

Due to the potential impacts of microplastics (MPs) and nanoplastics (NPs) on algal growth and thereby affect the climate-relevant substances, dimethylsulfoniopropionate (DMSP) and dimethyl sulfide (DMS), we studied the polystyrene (PS) MPs and NPs of 1 µm and 80 nm impacts on the growth, chlorophyll content, reactive oxygen species (ROS), antioxidant enzyme activity, and DMS/DMSP production in Emiliania huxleyi. E. huxleyi is a prominent oceanic alga that plays a key role in DMS and DMSP production. The results revealed that high concentrations of MPs and NPs inhibited the growth, carotenoid (Car), and Chl a concentrations of E. huxleyi. However, short-time exposure to low concentrations of PS MPs and NPs stimulated the growth of E. huxleyi. Furthermore, high concentrations of MPs and NPs resulted in an increase in the superoxide anion radical (O2.-) production rate and a decrease in the malondialdehyde (MDA) content compared with the low concentrations. Exposure to MPs and NPs at 5 mg L-1 induced superoxide dismutase (SOD) activity as a response to scavenging ROS. High concentrations of MPs and NPs significantly inhibited the production of DMSP and DMS. The findings of this study support the potential ecotoxicological impacts of MPs and NPs on algal growth, antioxidant system, and dimethylated sulfur compounds production, which maybe potentially impact the global climate.

Impact of environmental factors changes induced by marine heatwaves and heavy precipitation on antibiotic toxicity to Isochrysis galbana: Implications for climate change adaptation.

Isochrysis galbana, a crucial primary producer and food source in aquatic ecosystems, faces increasing challenges from climate change and emerging contaminants like antibiotics. This study investigates the combined effects of sudden temperature increase (representing marine heatwaves) and rapid salinity change (representing extreme precipitation events) on the toxicity of tetracycline (TC) and oxytetracycline (OTC) to I. galbana. Short-term experiments reveal heightened antibiotic toxicity at 31 °C or salinities of 18 PSU, surpassing algal tolerance limits. Long-term tests show decreased inhibition of algal growth on day 9, indicating algal adaptation to the environment. Analyses of photosynthesis II efficiency, pigment content, and macromolecular composition support this, suggesting adaptation mechanism activation. While algae acclimate to the environment during long-term antibiotic exposure, extreme weather conditions may compromise this adaptation. These findings have implications for managing antibiotics in aquatic environments under climate change.

A Novel Algicidal Bacterium, Microbulbifer sp. YX04, Triggered Oxidative Damage and Autophagic Cell Death in Phaeocystis globosa, Which Causes Harmful Algal Blooms.

Phaeocystis globosa causes severe marine pollution by forming harmful algal blooms and releasing hemolytic toxins and is therefore harmful to marine ecosystems and aquaculture industries. In this study, Microbulbifer sp. YX04 exerted high algicidal activity against P. globosa by producing and secreting metabolites. The algicidal activity of the YX04 supernatant was stable after exposure to different temperatures (-80 to 100°C) and pH values (4 to 12) for 2 h, suggesting that algicidal substances could temporarily be stored under these temperature and pH value conditions. To explore the algicidal process and mechanism, morphological and structural changes, oxidative stress, photosynthesis, autophagic flux, and global gene expression were investigated. Biochemical analyses showed that the YX04 supernatant induced reactive oxygen species (ROS) overproduction, which caused lipid peroxidation and malondialdehyde (MDA) accumulation in P. globosa. Transmission electron microscopy (TEM) observation and the significant decrease in both maximum photochemical quantum yield (Fv/Fm) and relative electron transfer rate (rETR) indicated damage to thylakoid membranes and destruction of photosynthetic system function. Immunofluorescence, immunoblot, and TEM analyses indicated that cellular damage caused autophagosome formation and triggered large-scale autophagic flux in P. globosa. Transcriptome analysis revealed many P. globosa genes that were differentially expressed in response to YX04 stress, most of which were involved in photosynthesis, respiration, cytoskeleton, microtubule, and autophagosome formation and fusion processes, which may trigger autophagic cell death. In addition to P. globosa, the YX04 supernatant showed high algicidal activity against Thalassiosira pseudonana, Thalassiosira weissflogii, Skeletonema costatum, Heterosigma akashiwo, and Prorocentrum donghaiense. This study highlights multiple mechanisms underlying YX04 supernatant toxicity toward P. globosa and its potential for controlling the occurrence of harmful algal blooms. IMPORTANCEP. globosa is one of the most notorious harmful algal bloom (HAB)-causing species, which can secrete hemolytic toxins, frequently cause serious ecological pollution, and pose a health hazard to animals and humans. Hence, screening for bacteria with high algicidal activity against P. globosa and studies on the algicidal characteristics and mechanism will contribute to providing an ecofriendly microorganism-controlling agent for preventing the occurrence of algal blooms and reducing the harm of algal blooms to the environment. Our study first reported the algicidal characteristic and mechanism of Microbulbifer sp. YX04 against P. globosa and demonstrated that P. globosa shows different response mechanisms, including movement ability, antioxidative systems, photosynthetic systems, gene expression, and cell death mode, to adapt to the adverse environment when algicidal compounds are present.

Interactive effects of increased temperature, elevated pCO2 and different nitrogen sources on the coccolithophore Gephyrocapsaoceanica.

As a widespread phytoplankton species, the coccolithophore Gephyrocapsaoceanica has a significant impact on the global biogeochemical cycle through calcium carbonate precipitation and photosynthesis. As global change continues, marine phytoplankton will experience alterations in multiple parameters, including temperature, pH, CO2, and nitrogen sources, and the interactive effects of these variables should be examined to understand how marine organisms will respond to global change. Here, we show that the specific growth rate of G. oceanica is reduced by elevated CO2 (1000 µatm) in [Formula: see text]-grown cells, while it is increased by high CO2 in [Formula: see text]-grown ones. This difference was related to intracellular metabolic regulation, with decreased cellular particulate organic carbon and particulate organic nitrogen (PON) content in the [Formula: see text] and high CO2 condition compared to the low CO2 condition. In contrast, no significant difference was found between the high and low CO2 levels in [Formula: see text] cultures (p > 0.05). The temperature increase from 20°C to 25°C increased the PON production rate, and the enhancement was more prominent in [Formula: see text] cultures. Enhanced or inhibited particulate inorganic carbon production rate in cells supplied with [Formula: see text] relative to [Formula: see text] was observed, depending on the temperature and CO2 condition. These results suggest that a greater disruption of the organic carbon pump can be expected in response to the combined effects of increased [Formula: see text]/[Formula: see text] ratio, temperature, and CO2 level in the oceans of the future. Additional experiments conducted under nutrient limitation conditions are needed before we can extrapolate our findings to the global oceans.

Effects of florfenicol on growth, photosynthesis and antioxidant system of the non-target organism Isochrysis galbana.

Florfenicol (FFC) is one of the most universally used antibiotics in aquaculture, which is substitute for chloramphenicol extensively, while the massive residues in aquatic environment were assumed to threaten the non-target organisms. Present research investigated the effects of florfenicol on growth, chlorophyll content, photosynthesis, and antioxidant ability of Isochrysis galbana. The results showed that FFC at 0.001-1 mg/L stimulated the growth of I. galbana and increased the content of chlorophyll. In addition, photosynthesis of I. galbana was inhibited and the photosynthetic parameters were uplifted with the increased exposure duration and FFC concentration. Furthermore, superoxide dismutase (SOD), catalase (CAT) activity significantly dropped at 0.01-20 mg/L FFC, while the contents of malondialdehyde (MDA), glutathione (GSH) and reactive oxygen species (ROS) increased after 72 h exposure, indicating that FFC at high concentrations caused a serious oxidative stress on algae. The simultaneous increase of ROS disrupted the equilibration between oxidants and antioxidant systems. Under the high concentration of FFC, the excessive of ROS was generated in algae which affected the membrane permeability and further decreased the cell biomass. Present study showed that acute exposure (72 h) at the environmental relevant concentration (0.01 mg/L) cannot induce the physiological dysfunction of the microalgae I. galbana, but the feeding concentration (20 mg/L) can. Additionally, this study hinted the possible negative impacts on ecosystems with the chronic exposure even at low FFC concentration or with the uncontrolled use of FFC.

The Antialgal Mechanism of Luteolin-7-O-Glucuronide on Phaeocystis globosa by Metabolomics Analysis.

Antialgal compounds from plants have been identified as promising candidates for controlling harmful algal blooms (HABs). In our previous study, luteolin-7-O-glucuronide was used as a promising algistatic agent to control Phaeocystis globosa (P. globose) blooms; however, its antialgal mechanism on P. globosa have not yet been elaborated in detail. In this study, a liquid chromatography linked to tandem mass spectrometry (LC-MS/MS)-based untargeted metabolomic approach was used to investigate changes in intracellular and extracellular metabolites of P. globosa after exposure to luteolin-7-O-glucuronide. Significant differences in intracellular metabolites profiles were observed between treated and untreated groups; nevertheless, metabolic statuses for extracellular metabolites were similar among these two groups. For intracellular metabolites, 20 identified metabolites showed significant difference. The contents of luteolin, gallic acid, betaine and three fatty acids were increased, while the contents of α-Ketoglutarate and acetyl-CoA involved in tricarboxylic acid cycle, glutamate, and 11 organic acids were decreased. Changes in those metabolites may be induced by the antialgal compound in response to stress. The results revealed that luteolin played a vital role in the antialgal mechanism of luteolin-7-O-glucuronide on P. globosa, because luteolin increased the most in the treatment groups and had strong antialgal activity on P. globosa. α-Ketoglutarate and acetyl-CoA were the most inhibited metabolites, indicating that the antialgal compound inhibited the growth through disturbed the tricarboxylic acid (TCA) cycle of algal cells. To summarize, our data provides insights into the antialgal mechanism of luteolin-7-O-glucuronide on P. globosa, which can be used to further control P. globosa blooms.

Growth Inhibition of Phaeocystis Globosa Induced by Luteolin-7-O-glucuronide from Seagrass Enhalus acoroides.

Enhalus acoroides (E. acoroides) is one of the most common species in seagrass meadows. Based on the application of allelochemicals from aquatic plants to inhibit harmful algal blooms (HABs), we used E. acoroides aqueous extract against harmful algae species Phaeocystis globosa (P. globosa). The results showed that E. acoroides aqueous extract could significantly inhibited the growth of P. globosa, decrease the chlorophyll-a content and photosynthetic efficiency (Fv/Fm) values of P. globosa, followed by vacuolization, plasmolysis, and the destruction of organelles. Twelve types of major chemical constituents were identified in E. acoroides aqueous extracts by ultraperformance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS), including six flavonoids, two homocyclic peptides, two long-chain aliphatic amides, one tannin, and one nitrogen heterocyclic compound. Flavonoids were the characteristic chemical constituents of E. acoroides aqueous extract. Furthermore, the antialgal activity of luteolin-7-O-glucuronide (68.125 µg/mL in 8 g/L E. acoroides aqueous extract) was assessed. The EC50-96 h value was 34.29 µg/mL. In conclusion, the results revealed that luteolin 7-O-glucuronide was one of the antialgal compounds of E. acoroides aqueous extract, with potential application as novel algaecide.

Inhibition effect of natural flavonoids on red tide alga Phaeocystis globosa and its quantitative structure-activity relationship.

Red tides that occur off coasts have become a worldwide phenomenon over the past decades. In order to mitigate the damage of the red tides on the aquatic ecosystems, it is crucial to develop a method for predicting algicidal activities that requires less labor and time, and most importantly, this method can quickly screen potential algicides to control red tides. In this study, we have investigated the algicidal activity of 19 natural flavonoids against a typical red tide alga, Phaeocystis globosa. Our results indicate that after 5 days of flavonoid exposure, the half inhibition concentrations (IC50) ranged from 0.068 to 3.065 mg L-1, which showed the strong algicidal activities of the flavonoids. Furthermore, quantitative structure activity relationship (QSAR) model has been carried out between negative scale logarithm (pIC50) of the flavonoids and the corresponding molecular descriptors. The developed model was validated, both internally and externally, which displayed statistical robustness (R2 = 0.867, p = 0.0002, Q2LOO = 0.825, RMSEC = 0.182, Q2extF3 = 0.896, RMSEP = 0.161, CCC = 0.935). This indicates that the developed model was obtained successfully with satisfactory predictability and robustness for the future rapid screening of natural flavonoids with high inhibition activity on the red tide alga growth. Moreover, the main descriptors in the QSAR model were the molar refractivity, partition coefficient, lowest unoccupied molecular orbital, and highest occupied molecular orbital, illustrating that the molecular electro-chemical characteristics are significant in the algicidal actions of the flavonoids. Graphical abstract Red tides frequently occur worldwide and have become a global environment problem. Flavonoids showed great potential in allelopathic control of the excessive growth of red tide algae. In this study, the algicidal activity of 19 natural flavonoids was investigated on a typical red tide organism Phaeocystis globosa. Futhermore, we applied the quantitative structure-activity relationship (QSAR) model to the experimental data. The model between molecular descriptors of flavonoids and their antialgal activity displays statistical robustness, and 4 of 45 selected molecular descriptors were obtained by regression of training set. The numbers in the figure represent the half inhibition concentration (IC50) of flavonoids. Our results show that the algicidal activity of flavonoids is closely related to molar refraction, partition coefficient, lowest unoccupied molecular orbital, and highest occupied molecular orbital. The QSAR model can efficaciously predict the algicidal activity and provide insights into the inhibitory mechanisms of flavonoids.

How haptophytes microalgae mitigate vitamin B12 limitation.

Vitamin B12 (cobalamin) can control phytoplankton development and community composition, with around half of microalgal species requiring this vitamin for growth. B12 dependency is determined by the absence of cobalamin-independent methionine synthase and is unrelated across lineages. Despite their important role in carbon and sulphur biogeochemistry, little is known about haptophytes utilization of vitamin B12 and their ability to cope with its limitation. Here we report the first evaluation of B12 auxotrophy among this lineage based on molecular data of 19 species from 9 families. We assume that all species encode only a B12-dependent methionine synthase, suggesting ubiquitous B12 auxotrophy in this phylum. We further address the effect of different B12 limitations on the molecular physiology of the model haptophyte Tisochrysis lutea. By coupling growth assays in batch and chemostat to cobalamin quantification and expression analyses, we propose that haptophytes use three strategies to cope with B12 limitation. Haptophytes may assimilate dissolved methionine, finely regulate genes involved in methionine cycle and B12 transport and/or limit B12 transport to the mitochondrion. Taken together, these results provide better understanding of B12 metabolism in haptophytes and represent valuable data for deciphering how B12-producing bacteria shape the structure and dynamics of this important phytoplankton community.

Toxicity of binary mixtures of pesticides to the marine microalgae Tisochrysis lutea and Skeletonema marinoi: Substance interactions and physiological impacts.

This study screened binary mixtures of pesticides for potential synergistic interaction effects on growth of the marine microalgae Tisochrysis lutea and Skeletonema marinoi. It also examined the single and combined effects of three of the most toxic substances on microalgal physiology. Single substances were first tested on each microalgal species to determine their respective EC50 and concentration-response relationships. The toxicity of six and seven binary mixtures was then evaluated in microplate experiments on the growth of T. lutea and S. marinoi, respectively, using two mixture modelling approaches: isobolograms and the MIXTOX tool, based on Concentration Addition (CA) or Independent Action (IA) models. Significant cases of antagonism (for both species) and synergism (for S. marinoi) were observed for the mixtures of isoproturon and spiroxamine, and isoproturon and metazachlor, respectively. These two mixtures, together with that of isoproturon and diuron, for which additivity was observed, were further studied for their impacts on the physiology of each species. Exposures were thus made in culture flasks at three concentrations, or concentration combinations for mixtures, selected to cause 25%, 50% and 75% growth rate inhibition. The effects of the selected pesticides singly and in combination were evaluated at three perceived effect concentrations on esterase metabolic activity, relative lipid content, cytoplasmic membrane potential and reactive oxygen species (ROS) content by flow cytometry, and on photosynthetic quantum yield (ϕ'M) by PAM-fluorescence. Isoproturon and diuron singly and in mixtures induced 20-40% decreases in ϕ'M which was in turn responsible for a significant decrease in relative lipid content for both species. Spiroxamine and metazachlor were individually responsible for an increase in relative lipid content (up to nearly 300% for metazachlor on S. marinoi), as well as cell depolarization and increased ROS content. The mixture of isoproturon and metazachlor tested on S. marinoi caused a 28-34% decrease in ϕ'M that was significantly higher than levels induced by each of substances when tested alone. This strong decrease in ϕ'M could be due to a combined effect of these substances on the photosynthetic apparatus, which is likely the cause of the synergy found for this mixture.

Interaction effects of oxytetracycline and copper at different ratios on marine microalgae Isochrysis galbana.

Contamination with both oxytetracycline (OTC) and Cu is prevalent in water. OTC can chelate with Cu to form OTC + Cu composites. Through the study of the effects of the interaction of OTC and Cu on the algae Isochrysis galbana at multiple coordination ratios, it was found that the OTC + Cu complex was antagonistic to algae growth after 24 h of exposure but was synergistic in treatment (T) 3 and T4 after 48 and 72 h. Compared with OTC alone, the addition of Cu addition significantly inhibited the biosynthesis of chlorophyll a, but the ratio (R) in the OTC + Cu and OTC treatments gradually increased from T1 to T4. The addition of Cu also led to a significant increase in malondialdehyde and reactive oxygen species, but R gradually decreased and increased, respectively, from T1 to T4. The accumulation of OTC in algae was considerably promoted by the addition of Cu, with R increasing 1.5 in T2 to 2.6 in T4; moreover, the residue of OTC in water was reduced in the presence of algae. OTC alone dramatically inhibited the absorption of Cu by algae, while in the presence of OTC + Cu, only the two high-OTC treatments showed a significant decrease in Cu absorption. In addition, the absorption of Mg was markedly inhibited in all OTC treatments and the adsorption of K in the high-OTC treatment, but these inhibitory effects were alleviated in the OTC + Cu treatment. These results indicated that the effects of the OTC + Cu complex on algae were different from the effects of OTC and Cu alone.

Effect of microplastics on the toxicity of chlorpyrifos to the microalgae Isochrysis galbana, clone t-ISO.

It is highly likely that phytoplanktonic organisms will interact with MPs in the ocean, and consequently with the pollutants sorbed onto their surfaces. Microalgae play an essential role in maintaining the balance of the marine ecosystem due to the fact that they are a primary producer and the base of marine trophic chains. Therefore, their fitness represents an important index in the assessment of water quality. The objectives of this study were i) to assess the toxicity of MPs and the pesticide chlorpyrifos (CPF) to the microalgae, Isochrysis galbana, clone t-ISO and ii) to ascertain whether the presence of MPs affects the toxicity of CPF. Microalgae growth rate was selected as the endpoint and a commercial virgin PE micronized powder was chosen as a micro-plastic model, with mean size ranging from 2 to 6 µm, assayed until 25 mg L-1. CPF was tested at concentrations ranging from 0 to 4 mg L-1. A constant concentration of MPs (5 mg L-1) was loaded with increasing doses of CPF (0-3 mg L-1) with a 2 h incubation period. Bioassays were performed at 20 °C, in glass tubes of 50 ml, with air and constant light and an exposure time of 72 h. Cell counts were performed using a Coulter Counter Multisizer III and HPLC was used to quantify the partition of this pollutant among MPs and water. Although microalgae growth was not impacted by MPs, growth was clearly affected by exposure to CPF from 2 mg L-1 and above, with a total growth inhibition at concentrations over 3 mg L-1. Subsequent to incubation, 80% of CPF was sorbed onto MP surfaces. Two different dose-response curves resulted from CPF bioassays depending on the presence of MP, with lower percentages of inhibition when CPF was presented through MP. Thus, the adsorption of CPF onto MP surfaces modulates the toxicity of CPF on I. galbana growth through a reduction in its toxicity, as CPF is adsorbed onto MP surfaces which are less bio-available to the algal cells.

Natural iron fertilization of the coastal ocean by "blackwater rivers".

The present study elucidates the role of natural iron fertilization of the coastal ocean by so-called "blackwater rivers". Areas of marsh, fen, peatland, boreal forest etc. are characterized by organic-rich soils. From those soils, humic substances (humic and fulvic acids) are leached to the aquatic system resulting in river water that is low in pH and dark-brown in color. The point is that "blackwater rivers" tend to be rich in dissolved iron due to the unique chelating properties of humic and fulvic acids which bind Fe(III) and keep it in solution. We performed algal physiological (growth rate) experiments under conditions of iron deficiency with the marine unicellular phytoplankton algae Chlorella salina and Diacronema lutheri in 0.2 µm cut-off filtered mixtures of natural "blackwater river" water and synthetic seawater. Our results demonstrate that the iron naturally present in "blackwater rivers" is readily bioavailable to both marine algal species. Furthermore, the humic and fulvic acids exert an additional stimulatory effect on the marine algae. Both algae thrive much better in the presence of natural humic and fulvic acids as compared to a medium where EDTA is used as an iron-chelating agent. Our results indicate that "blackwater rivers", in sharp contrast to other types of rivers, are excellent sources of bioavailable iron to marine phytoplankton. This natural iron fertilization may give rise to photosynthesis-driven sequestration of CO2 from the atmosphere to the sea, as can be seen from the visualization of CO2 surface concentrations by NASA (NASA GEOS-5 model) which shows the global sources and sinks of CO2 localized in time and space. The results by NASA suggest that strong marine CO2 sinks in coastal waters tend to occur close to "blackwater river" estuaries. It is thus evident that "blackwater rivers" act as important sources of a limiting nutrient (iron) to the ocean.

Nitric oxide production and antioxidant function during viral infection of the coccolithophore Emiliania huxleyi.

Emiliania huxleyi is a globally important marine phytoplankton that is routinely infected by viruses. Understanding the controls on the growth and demise of E. huxleyi blooms is essential for predicting the biogeochemical fate of their organic carbon and nutrients. In this study, we show that the production of nitric oxide (NO), a gaseous, membrane-permeable free radical, is a hallmark of early-stage lytic infection in E. huxleyi by Coccolithoviruses, both in culture and in natural populations in the North Atlantic. Enhanced NO production was detected both intra- and extra-cellularly in laboratory cultures, and treatment of cells with an NO scavenger significantly reduced viral production. Pre-treatment of exponentially growing E. huxleyi cultures with the NO donor S-nitroso-N-acetylpenicillamine (SNAP) prior to challenge with hydrogen peroxide (H2O2) led to greater cell survival, suggesting that NO may have a cellular antioxidant function. Indeed, cell lysates generated from cultures treated with SNAP and undergoing infection displayed enhanced ability to detoxify H2O2. Lastly, we show that fluorescent indicators of cellular ROS, NO, and death, in combination with classic DNA- and lipid-based biomarkers of infection, can function as real-time diagnostic tools to identify and contextualize viral infection in natural E. huxleyi blooms.

Low-dose stimulation of growth of the harmful alga, Prymnesium parvum, by glyphosate and glyphosate-based herbicides.

Glyphosate-based herbicides (GBH) are widely used around the globe. While generally toxic to phototrophs, organic phosphorus in glyphosate can become available to glyphosate-resistant phytoplankton and contribute to algal bloom development. Few studies have examined the effects of GBH on growth of eukaryotic microalgae and information for the toxic bloom-forming haptophyte, Prymnesium parvum, is limited. Using a batch-culture system, this study examined the effects on P. parvum growth of a single application of Roundup Weed and Grass Killer Super Concentrate Plus® (Roundup SC), Roundup Weed and Grass Killer Ready-to-Use III® (Roundup RtU), and technical-grade glyphosate at low concentrations [0-1000 µg glyphosate acid equivalent (ae) l-1]. Roundup formulations differ in the percent of glyphosate as active ingredient (Roundup SC, ∼50%; Roundup RtU, 2%), allowing indirect evaluation of the influence of inactive ingredients. Roundup SC enhanced exponential growth rate at 10-1000 µg glyphosate ae l-1, and a positive monotonic association was noted between Roundup SC concentration and early (pre-exponential growth) but not maximum cell density. Glyphosate and both Roundup formulations enhanced growth rate at 100 µg glyphosate l-1, but only Roundup SC and glyphosate significantly stimulated early and maximum density. This observation suggests the higher concentration of inactive ingredients and other compounds in Roundup RtU partially counteracts glyphosate-dependent growth stimulation. When phosphate concentration was varied while maintaining other conditions constant, addition of Roundup SC and glyphosate at 100 µg l-1 influenced growth more strongly than equivalent changes in phosphate-associated phosphorus. It appears, therefore, that low doses of glyphosate stimulate growth by mechanisms unrelated to the associated small increases in total phosphorus. In conclusion, glyphosate and GBH stimulate P. parvum growth at low, environmentally relevant concentrations. This finding raises concerns about the potential contribution to P. parvum blooms by glyphosate-contaminated runoff or by direct application of GBH to aquatic environments.

Investigation of the Inhibitory Effects of Mangrove Leaves and Analysis of Their Active Components on Phaeocystis globosa during Different Stages of Leaf Age.

The presence of harmful algal blooms (HABs) can cause significant problems to the quality of the water, the marine ecosystems, and the human health, and economy worldwide. Biological remediation can inhibit harmful algal growth efficiently in an environmental-friendly manner. Therefore, the research conducted on biological remediation with regard to the inhibition of HABs is becoming a major focus in marine ecology. To date, no study has been reported with regard to the red tides occurring in mangrove wetlands. Therefore, the present study used two mangrove species, namely Bruguiera gymnorrhiza and Kandelia candel and one harmful algae species Phaeocystis globosa as experimental organisms. The present study determined the inhibitory effects and algae physiology of specific aqueous extracts from mangrove leaves on the viability of harmful algae, and analyzed the main chemical composition of the aqueous extracts by ultra-performance liquid chromatography coupled to high resolution mass spectrometry (UPLC-QTOF-MS). The results indicated that the aqueous extracts from different leaf ages of B. gymnorrhiza and K. candel leaves exhibited apparent inhibitory effects on the growth of P. globosa. The inhibitory effects of B. gymnorrhiza and K. candel leaves aqueous extracts on the growth of P. globosa were in the following order: senescent > mature > young leaves. The levels of the parameters superoxide dismutase (SOD) activity, glutathione (GSH), and malondialdehyde (MDA)content in P. globosa following treatment with B. gymnorrhiza and K. candel leaves aqueous extracts were increased as follows: senescent > mature > young leaves. Simultaneously, the intensity of the ion peaks of the specific secondary metabolites assigned 4 (No.: 4 Rt: 2.83 min), 7 (No.: 7 Rt: 3.14 min), 8 (No.: 8 Rt: 3.24 min), 9 (No.: 9 Rt: 3.82min) and 10 (No.: 10 Rt: 4.10 min) were increased. These metabolites were found in the aqueous extracts from B. gymnorrhiza leaves. The intensities of the ion peaks of the secondary metabolites 7, 8 in the aqueous extracts from the K. candel leaves were also increased. The majority of the substances that inhibited the algae found in the mangrove plants were secondary metabolites. Therefore, we considered that the norsesquiterpenes compounds 4, 8, 9, and 10 and a phenolic glycoside compound 7 were the active constituents in the aqueous extracts of the mangrove leaves responsible for the inhibition of algae growth. This evidence provided theoretical guidance for the development of biological methods to control red tides and for the further use of substances with antiproliferative activity against algae.

The Response of Three Southern Ocean Phytoplankton Species to Ocean Acidification and Light Availability: A Transcriptomic Study.

Ocean acidification (OA) and high light was found to negatively affect the Antarctic key species Phaeocystis antarctica, Fragilariopsis kerguelensis and Chaetoceros debilis. To unravel the underlying physiological response at the transcriptomic level, these species were grown under ambient and elevated pCO2 combined with low or high light. RNA sequencing revealed that the haptophyte was much more tolerant towards OA than the two diatoms as only these showed distinct OA-dependent gene regulation patterns. Under ambient pCO2, high light resulted in decreased glycolysis in P. antarctica. Contrastingly, upregulation of genes related to cell division and transcription as well as reduced expression of both cata- and anabolic carbon related pathways were seen in C. debilis. OA in combination with low light led to reduced respiration, but also surprisingly to higher expression of genes involved in light protection, transcription and translation in C. debilis. Though not affecting P. antarctica, OA combined with high light caused also photosensitivity in both diatoms. As additional response reallocation of carbon to lipids was found in C. debilis under these conditions. Overall, we conclude that P. antarctica is better adapted than the two diatoms to OA and high light.