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The study aimed to assess the effects of nine combinations of phytohormones, salicylic acid (SA), gibberellic acid (GA), and jasmonic acid (JA) on the growth, physiology, and biochemistry of Aurantiochytrium sp. Parameters like optical density (OD), biomass, protein content, hydrogen peroxide (H 2 O 2 ), malondialdehyde (MDA), catalase activity (CAT), and gene expression (malic enzyme (ME) and acetyl-CoA carboxylase (ACCase)) were assessed at various cultivation stages (24, 48, 72, and 96 h). The research also analyzed fatty acid composition, unsaturated fatty acids (UFA), saturated fatty acids (SFA), and the UFA to SFA ratio (USS) to understand the biochemical changes induced by phytohormones. Results demonstrated that modifying phytohormone concentrations significantly affected the characteristics of the microalgae, particularly in correlation with different growth stages, emphasizing the necessity of precise control of phytohormone levels for optimizing cultivation conditions and enhancing bioactive compound production in Aurantiochytrium sp., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Aurantiochytrium is a well-known long-chain polyunsaturated fatty acids (PUFAs) producer, especially docosahexaenoic acid (DHA). In order to reduce the cost or improve the productivity of DHA, many researchers are focusing on exploring the high-yield strain, reducing production costs, changing culture conditions, and other measures. In this study, DHA production was improved by a two-stage fermentation. In the first stage, efficient and cheap soybean powder was used instead of conventional peptone, and the optimization of fermentation conditions (optimal fermentation conditions: temperature 28.7 °C, salinity 10.7‱, nitrogen source concentration 1.01 g/L, and two-nitrogen ratio of yeast extract to soybean powder 2:1) based on response surface methodology resulted in a 1.68-fold increase in biomass concentration. In the second stage, the addition of 2.5 mM sesamol increased the production of fatty acid and DHA by 93.49% and 98.22%, respectively, as compared to the optimal culture condition with unadded sesamol. Transcriptome analyses revealed that the addition of sesamol resulted in the upregulation of some genes related to fatty acid synthesis and antioxidant enzymes in Aurantiochytrium . This research provides a low-cost and effective culture method for the commercial production of DHA by Aurantiochytrium sp.

To compare the influence of water samples collected from various areas on toxic effect of HNS, we examined the toxic effect of two commonly found HNS: p-chloroaniline and butyl acrylate, on Nannochloropsis oculata cultured in seawater collected from Laizhou bay and Jiaozhou bay (China). The results showed that both p-chloroaniline and butyl acrylate had significant toxic effect on N. oculata cultured in both water samples. P-chloroaniline inhibited its net oxygenation rate and oxygen consumption rate. Butyl acrylate inhibited the net oxygenation rate whereas significantly stimulated oxygen consumption rate. Performance of N. oculata changed between two water samples under same level of p-chloroaniline and butyl acrylate. The net oxygenation rate of N. oculata cultured in the seawater from the Jiaozhou bay increased by 11.60 %, the oxygen consumption rate increased by 26.91 %, algae cell growth decreased by 16.83 %, compared to those from Laizhou bay. The Fv/Fm of N. oculata cultured in Jiaozhou bay was more significantly inhibited at 0.5 and 2.0 mg L -1 p-chloroaniline, while it was significantly inhibited at 5. 0 mg L -1 of butyl acrylate, compared to those from Laizhou bay. Moreover, the toxic effect of both HNS on net oxygenation rate and oxygen consumption rate were significantly attenuated as the concentration increased. The growth inhibition of microalgae cultured in two seawater samples was more evident at 0.5 and 5.0 mg L -1 p-chloroaniline than at 2.0 mg L -1 p-chloroaniline,and the growth inhibition of microalgae cultured in two seawater samples was more evident at 2.0 and 5.0 mg L -1 butyl acrylate than at 0.5 mg L -1 butyl acrylate. These results indicated that toxic effect of p-chloroaniline and butyl acrylate on the growth of N. oculata was influenced by the pollutants in the two water samples. Consequently, a corresponding research on water sample is required in advance to increase accuracy of future ecological risk assessment of HNS., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The toxic alga Heterosigma akashiwo (Raphidophyceae) is known to form harmful algal blooms (HABs), which can have serious negative effects on the aquatic ecosystem and human life. Previous study has shown that Nω-acetylhistamine (N-AcH), an algicidal compound secreted by algicidal bacteria Bacillus sp. Strain B1, can inhibit the growth of H. akashiwo. In this study, the algicidal mechanism of N-AcH against H. akashiwo was explored, and the changes of toxicity of H. akashiwo treated with N-AcH were investigated. The algal inhibition rate was calculated by the optical density method, and the results showed that the growth inhibition rate of H. akashiwo was about 90% when treated in the medium with 40 μg/mL N-AcH at 96 h. After 72 h treatment, transmission electron microscopy (TEM) showed that the microstructure of H. akashiwo cell was seriously damaged at this concentration. The content of Chlorophyll a and Chlorophyll b decreased while malonaldehyde levels increased, and superoxide dismutase activity first increased and then decreased as well as soluble protein content. GC-MS revealed that the type and content of fatty acids cut down after 48 h and 96 h treatment. Hemolytic test, MTT assay, and micronucleus test all demonstrated the decrease in the toxicity of H. akashiwo treated with 40 μg/mL N-AcH. In brief, N-AcH mainly kills H. akashiwo cell through oxidative stress and can also reduce its toxicity, so it is a promising algicide with the dual functions of killing algae and inhibiting algal toxic effects., (Copyright © 2020. Published by Elsevier Inc.)

Bioassay-guided separation of a methanol extract of Tricleocarpa jejuensis by monitoring algicidal activity against the red tide phytoplankton Chattonella antiqua led to the isolation of an active fraction consisting of a mixture of four isomeric compounds. The active compounds were identified as (E)-9-hydroxyoctadec-10-enoic acid (1), (E)-10-hydroxyoctadec-8-enoic acid (2), (E)-11-hydroxyoctadec-12-enoic acid (3) and (E)-12-hydroxyoctadec-10-enoic acid (4) by NMR, IR and mass spectral data. The structures were confirmed by comparison of the NMR and MS data with those of authentic samples of 1-4 obtained by unambiguous syntheses. Synthesized hydroxy acids 1-4 and related compounds were assessed for algicidal activity against C. antiqua and it was found that all of 1-4 had high activity (>80% mortality at 24 h) at a concentration of 20 μg/mL. A structure-activity relationship study using 11 related compounds revealed that the presence of the hydroxyl group is important for the activity and the double bond may be replaced with a triple bond., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Effect of water accommodated fractions (WAF) of #180 fuel oil on fixed carbon and nitrogen in microalgae was studied by stable isotopes. Platymonas helgolandica, Heterosigma akashiwo and Nitzschia closterium were exposed to five WAF concentrations for 96 h. The δ 13 C value of microalgae was significantly lower than that of the control group, indicated that carbon was limited in the WAF concentrations. The δ 13 C value of microalgae appeared peak valley at 48 h in control group, corresponding to the enhanced capacity in carbon fixation during microalgae photosynthesis. The physiological acclimation capacity of microalgae was revealed by the occurrence time when the δ13C value was in peak valley, and thus the physiological acclimation capacity of microalgae decreased in the order of Nitzschia closterium > Heterosigma akashiwo > Platymonas helgolandica. Principal component analysis (PCA) were applied to the δ13C value in order to verify the "hormesis" phenomenon in microalgae. The δ13C value could discriminate between stimulatory effects at low doses and inhibitory effects at high doses. In addition, the present study also investigated the effect of the nitrogen on microalgae growth. Because microalgae could still absorb the NO 3 -N and release of NO 2 -N and NH 4 -N in present study, the nitrogen cycle in microalgae was in the equilibrium status. The δ 15 N value in microalgae exhibited no obvious change with the increasing of WAF concentrations at the same time. However, due to the enrichment of nitrogen, the δ 15 N value first increased gradually with the time and finally was stable. Overall, the fractionation of carbon and nitrogen stable isotopes illustrated that the effect of carbon on the growth of microalgae was more prominent than nitrogen. Stable isotopes was used to investigate the influence of WAF on fixed carbon and nitrogen in microalgae growth, providing a fundamental theoretical guidance for risk assessment of marine ecological environment., Competing Interests: Declaration of competing interest Submission of an article implies that the work described has not been published previously (except in the form of an abstract or as part of a published lecture or academic thesis), that it is not under consideration for publication elsewhere, that its publication is approved by all authors and tacitly or explicitly by the responsible authorities where the work was carried out, and that, if accepted, it will not be published elsewhere in the same form, in English or in any other language, without the written consent of the copyright-holder. By attaching this declaration to the submission, the corresponding author certifies that:, (Copyright © 2020 Elsevier Inc. All rights reserved.)

Harmful algal blooms (HABs) occur all over the world, producing severely negative effects on human life as well as on marine ecosystems. The algicidal compound, prodigiosin, secreted by algicidal bacteria Hahella sp. KA22 can lyse the harmful alga Heterosigma akashiwo. This study is aimed to investigate the algicidal mechanism of prodigiosin against H. akashiwo by detecting physiological and morphological responses of H. akashiwo to presence of prodigiosin. The results indicated that prodigiosin showed strong algicidal effects on H. akashiwo at the concentration of 3 μg/mL. Chlorophyll a and protein levels of the microalgae decreased significantly while malonaldehyde levels increased at this concentration. Contents of ascorbic acid and activities of superoxide dismutase and peroxidase increased fast with the quick decrease of the reactive oxygen species (ROS). For the 3 μg/mL prodigiosin treatment group, transcription of genes related to photosynthesis and respiration were significantly inhibited at 12 h while respiration related genes increased at 24 h. Collectively, the results indicated that prodigiosin could kill the microalgae by inducing ROS overproduction which could destroy the cell integrity and change the antioxidant system levels and functional gene expression. Our results demonstrated that prodigiosin is an effective algicide for the control of harmful algae., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, the impact of alpha-iron oxide (α-Fe 2 O 3 , 20-40 nm) and gamma iron oxide (γ-Fe 2 O 3 , 20-40 nm) nanoparticles (NPs) on phytoplankton species Selenastrum capricornutum and Nannochloropsis oculata was investigated Characterizations of the NPs were systematically carried out by TEM, dynamic light scattering, zeta potential, X-ray diffraction, SEM, and Fourier transformation infrared spectroscopy. Acute toxicity was tested between 0.2 and 50 mg/L for each NP for a period of 72 hours exposure. γ-Fe 2 O 3 NP inhibited development of N oculata at the rate of 54% in 0.2 mg/L group with a high mortality rate of up to 82%. α-Fe 2 O 3 NPs were less toxic that induced 97% mortality on N oculata at 10 mg/L suspensions. In contrast, α-Fe 2 O 3 NP inhibited growth of S capricornutum strongly (73%) in 0.2 mg/L group. γ-Fe 2 O 3 NPs showed similar growth inhibition (72%) on S capricornutum in 10 mg/L suspensions. Despite the differential effects, the results indicated acute toxicity of α-Fe 2 O 3 and γ-Fe 2 O 3 NPs on N oculata and S capricornutum., (© 2019 Wiley Periodicals, Inc.)

Background: Schizochytrium has been widely used in industry for synthesizing polyunsaturated fatty acids (PUFAs), especially docosahexaenoic acid (DHA). However, unclear biosynthesis pathway of PUFAs inhibits further production of the Schizochytrium. Unsaponifiable matter (UM) from mevalonate pathway is crucial to cell growth and intracellular metabolism in all higher eukaryotes and microalgae. Therefore, regulation of UM biosynthesis in Schizochytrium may have important effects on fatty acids synthesis. Moreover, it is well known that UMs, such as squalene and β-carotene, are of great commercial value. Thus, regulating UM biosynthesis may also allow for an increased valuation of Schizochytrium., Results: To investigate the correlation of UM biosynthesis with fatty acids accumulation in Schizochytrium, fluconazole was used to block the sterols pathway. The addition of 60 mg/L fluconazole at 48 h increased the total lipids (TLs) at 96 h by 16% without affecting cell growth, which was accompanied by remarkable changes in UMs and NADPH. Cholesterol content was reduced by 8%, and the squalene content improved by 45% at 72 h, which demonstrated fluconazole's role in inhibiting squalene flow to cholesterol. As another typical UM with antioxidant capacity, the β-carotene production was increased by 53% at 96 h. The increase of squalene and β-carotene could boost intracellular oxidation resistance to protect fatty acids from oxidation. The NADPH was found to be 33% higher than that of the control at 96 h, which meant that the cells had more reducing power for fatty acid synthesis. Metabolic analysis further confirmed that regulation of sterols was closely related to glucose absorption, pigment biosynthesis and fatty acid production in Schizochytrium., Conclusion: This work first reported the effect of UM biosynthesis on fatty acid accumulation in Schizochytrium. The UM was found to affect fatty acid biosynthesis by changing cell membrane function, intracellular antioxidation and reducing power. We believe that this work provides valuable insights in improving PUFA and other valuable matters in microalgae.

The toxic effects of p-xylene, 4-methylbenzyl alcohol, p-methyl benzaldehyde, and p-toluic acid on two marine microalgae (Phaeodactylum tricornutum Bohlin, and Skeletonema costatum) were investigated. p-Xylene was the most toxic to Pha. tricornutum with a 96 h EC 50 value of 6.76 mg L -1 . Based on the 96 h EC 50 values for two microalgae, the toxicity of the four chemicals, in descending order, was: p-xylene, p-methyl benzaldehyde, 4-methylbenzyl alcohol, then p-toluic acid. The results showed that the toxicity of the transformed products of p-xylene was lower than that of p-xylene.

The phytohormone 6-benzylaminopurine (6-BAP) significantly improves lipid synthesis of oleaginous microorganisms with the great potential applied in lipid production. In the current study, the lipid and DHA productions in oleaginous Aurantiochytrium sp. were found to be improved by 48.7% and 55.3%, respectively, induced by 6-BAP treatments. Then, using high-throughput RNA-seq technology, the overall de novo assembly of the cDNA sequence data generated 53871 unigenes, and 15902 of these were annotated in at least one database. The comparative transcriptomic profiles of cells with and without 6-BAP treatments revealed that a total of 717 were differently expressed genes (DE), with 472 upregulated and 245 downregulated. Further annotation and categorization indicated that some DE genes were involved in pathways crucial to lipid and DHA productions, such as fatty acid synthesis, central carbon metabolism, transcriptional factor, signal transduction, and mevalonate pathway. A regulation mode of 6-BAP, in turn, perception and transduction of 6-BAP signal, transcription factor, expression regulations of the downstream genes, and metabolic changes, respectively, was put forward for the first time in the present study. This research illuminates the transcriptomic mechanism of phytohormone stimulation of lipid and DHA production in an oleaginous microorganism and provides the potential targets modified using genetic engineering for improving lipid and DHA productivity.

Total lipids and docosahexaenoic acid (DHA) production by a Chilean isolated thraustochytrid were evaluated under different growth conditions in shake flasks. The analyzed strain was identified as Thraustochytrium striatum according to an 18S rRNA gene sequence analysis. The strain (T. striatum AL16) showed negligible growth in media prepared with artificial seawater at concentrations lower than 50% v/v and pH lower than 5. Maltose and starch were better carbon sources for growth than glucose. DHA content of the biomass grown with maltose (60 g L -1 ) was doubled by increasing the agitation rate from 150 to 250 rpm. The DHA (0.8-6%) and eicosapentaenoic acid (0.2-21%) content in the total lipids varied depending on culture conditions and culture age. Lipid and DHA concentration increased (up to 5 g L -1 and 66 mg L -1 , respectively) by regularly feeding the culture with a concentrated starch solution. Carotenoid accumulation was detected in cells grown with maltose or starch. Contrasting conditions of starch and glucose cultures were selected for comparative proteomics. Total protein extracts were separated by two-dimensional gel electrophoresis; 25 spots were identified using ESI-MS/MS. A protein database (143,006 entries) for proteomic interrogation was generated using de novo assembling of Thraustochytrium sp. LLF1b - MMETSP0199_2 transcriptome; 18 proteins differentially expressed were identified. Three ATP synthases were differentially accumulated in cultures with glucose, whereas malate dehydrogenase was more abundant in cells cultured with starch.

Microalgae biorefinery is presently receiving a lot of attention as driven by its production of high value-added products. In this study, an oleaginous microalga Aurantiochytrium limacinum SR21 was cultured for docosahexaenoic acid (DHA) production using 20% (w/v) of K 2 HPO 4 -waste feedstock to replace 0.005% (w/v) of KH 2 PO 4 in the flask culture. DHA is an essential nutrient for human's brain functionalities. Collectively, the K 2 HPO 4 -waste feedstock with working concentration of 0.005% (w/v) in the cultivation prompted a higher lipid content (8.29%) and DHA production (128.81 mg.L -1 ). Moreover, natural plant pigment products containing stabilised betacyanins were utilised as natural red colourants for hard candy production. This study develops microalgal cultivation using salt-rich waste feedstock for a higher lipid and DHA content as well as application of natural colouring agents in food products., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Nine antialgal active compounds, (i.e. trehalose (1), twenty-two methyl carbonate (2), (-)-dihydromenisdaurilide (3), 3,7,11,15-tetramethyl-2-hexadecen-1-ol (4), isophytol (5), 8-hexadecenol (6), 17-hydroxyheptadecanoic acid (7), trans-asarone (8) and 2-amino-3-mercaptopropanoic acid (9)) were isolated from Ulva pertusa for the first time by sephadex LH-20 column chromatography, silica gel column chromatography and repeated preparative TLC. Except for compound 4, all compounds represented novel isolated molecules from marine macroalgae. Further, antialgal activities of these compounds against Amphidinium carterae, Heterosigma akashiwo, Karenia mikimitoi, Phaeocystis globosa, Prorocentrum donghaiense and Skeletonema costatum were investigated for the first time. Results showed these nine compounds have selectivity antialgal effects on all test red tide microalgae, and antialgal activities against red tide microalgae obviously enhanced with the increase of concentration of antialgal compounds. Based on this, EC 50-96 h values of these nine compounds for six red tide microalgae were obtained for the first time. By analyzing and comparing EC 50-96 h values, it has been determined that seven compounds (1, 3, 4, 6, 7, 8 and 9) showed the superior application potential than potassium dichromate or gossonorol and other six compounds as a characteristic antialgal agent against Heterosigma akashiwo, Karenia mikimitoi and Prorocentrum donghaiense. Overall this study has suggested that green algae Ulva pertusa is a new source of bioactive compounds with antialgal activity., (Copyright © 2018. Published by Elsevier Inc.)

Increasing concern has been focused on the potential risks associated with the trophic transfer to aquatic organisms of ambient contaminants in the presence of titanium dioxide nanoparticles (nano-TiO 2 ). This study investigated the influence of nano-TiO 2 on the trophic transfer of arsenic (As) from the microalgae Nannochloropsis maritima to the brine shrimp Artemia salina nauplii. We found that nano-TiO 2 could significantly facilitate As sorption on N. maritima within an exposure period of 24 h, and this sorption subsequently led to higher As trophic transfer from the algae to A. salina according to trophic transfer factors (TTF As +nano-TiO2  > TTF As ). However, after 48 h of depuration, the retention of As in A. salina fed As-nano-TiO 2 -contaminated algae was even lower than that in A. salina fed As-contaminated algae at the same exposure concentrations. This result indicates that the increased food chain transfer of As in the presence of nano-TiO 2 can be explained by adsorption of As onto nano-TiO 2 in contaminated food (algae), but the bioavailability of As in A. salina is reduced after the introduction of nanoparticles. Although the stress enzyme activities of superoxide dismutase (SOD) and acetylcholinesterase (AChE) in A. salina at a lower As concentration treatment in the presence of nano-TiO 2 were not significantly changed, they increased with higher exposure concentrations of As with or without nano-TiO 2 . Our study highlighted the complex role of nanomaterials in the transfer of ambient contaminants via trophic chains and the potential of nano-TiO 2 to reduce the bioavailability of As via trophic transfer to saltwater zooplankton., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Schizochytrium sp. accumulates valuable polyunsaturated fatty acid (PUFA), such as docosahexaenoic acid (DHA). In order to increase DHA synthesis in this microorganism, physical or chemical mutagenesis aided with powerful screening methods are still preferable, as its DHA synthetic pathway has not yet been clearly defined for gene manipulation. To breed this agglomerate microorganism of thick cell wall and rather large genome for increasing lipid content and DHA percentage, a novel strategy of atmospheric and room temperature plasma (ARTP) mutagenesis coupled with stepped malonic acid (MA) and zeocin resistance screening was developed. The final resulted mutant strain mz-17 was selected with 1.8-fold increased DHA production. Accompanied with supplementation of Fe 2+ in shake flask cultivation, DHA production of 14.0 g/L on average was achieved. This work suggests that ARTP mutation combined with stepped MA and zeocin resistance screening is an efficient method of breeding Schizochytrium sp. of high DHA production, and might be applied on other microorganisms for obtaining higher desired PUFA products.

Schizochytrium is a microalgae-like fungus and is widely used for producing docosahexaenoic acid (DHA). It is also a promising source of squalene and carotenoids. However, few fermentation strategies are available in enhancing squalene and carotenoid content in Schizochytrium. This study showed that butanol addition had multiple effects on Schizochytrium limacinum B4D1. First, butanol addition altered the lipid content of cells. Second, 6g/L of butanol decreased the proportion of DHA by nearly 40%. Third, the squalene content increased 31-fold in the presence of 6g/L butanol. Finally, cells accumulated more carotenoids upon butanol addition. Specifically, when cells were treated with 8g/L butanol, the astaxanthin content increased to 245 times than that of the untreated control. These results are helpful for the commercial exploitation of Schizochytrium in producing squalene and carotenoids., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Docosapentaenoic acid/docosahexaenoic acid ratio (DPA/DHA ratio) in Schizochytrium was relatively stable. But ideally the ratio of DPA/DHA will vary according to the desired end use. This study reports several ways of modulating the DPA/DHA ratio. Incubation times changed the DPA/DHA ratio, and changes in this ratio were associated with the variations in the saturated fatty acid (SFAs) content. Propionic acid sharply increased the SFAs content in lipids, dramatically decreased the even-chain SFAs content, and reduced the DPA/DHA ratio. Pentanoic acid (C5:0) and heptanoic acid (C7:0) had similar effects as propionic acid, whereas butyric acid (C4:0), hexanoic acid (C6:0), and octanoic acid (C8:0) did not change the fatty acid profile and the DPA/DHA ratio. Transcription analyses show that β-oxidation might be responsible for this phenomenon. Iodoacetamide upregulated polyunsaturated fatty acid (PUFA) synthase genes, reduced the DHA content, and improved the DPA content, causing the DPA/DHA ratio to increase. These results present new insights into the regulation of the DPA/DHA ratio.

A European round robin test according to ISO 5725-2 was conceptually prepared, realised, and evaluated. The aim was to determine the inter-laboratory variability of the overall process for the ecotoxicological characterization of construction products in eluates and bioassays. To this end, two construction products BAM-G1 (granulate) and HSR-2 (roof sealing sheet), both made of EPDM polymers (rubber), were selected. The granular construction product was eluted in a one stage batch test, the planar product in the Dynamic Surface Leaching test (DSLT). A total of 17 laboratories from 5 countries participated in the round robin test: Germany (12), Austria (2), Belgium (1), Czech Republic (1) and France (1). A test battery of four standardised ecotoxicity tests with algae, daphnia, luminescent bacteria and zebrafish eggs was used. As toxicity measures, EC50 and LID values were calculated. All tests, except the fish egg test, were basically able to demonstrate toxic effects and the level of toxicity. The reproducibility of test results depended on the test specimens and the test organisms. Generally, the variability of the EC50 or LID values increased with the overall level of toxicity. For the very toxic BAM-G1 eluate a relative high variability of CV = 73%-110% was observed for EC50 in all biotests, while for the less toxic HSR-2 eluate the reproducibility of EC50 varied with sensitivity: it was very good (CV = 9.3%) for the daphnia test with the lowest sensitivity, followed by the algae test (CV = 36.4%). The luminescent bacteria test, being the most sensitive bioassay for HSR-2 Eluate, showed the highest variability (CV = 74.8%). When considering the complex overall process the reproducibility of bioassays with eluates from construction products was acceptable., (Copyright © 2017. Published by Elsevier Ltd.)

Fish-killing harmful algal blooms (HABs) of Chattonella marina causes serious hazards and risks to fish farming and environment throughout the world. At present, it is necessary to explore cost-effective and recyclable materials for controlling C. marina blooms to reduce the cost and control the potential side effect to the environment. A novel earth-abundant natural magnetic sphalerite (NMS) for removing C. marina was systematically investigated, including the effect of NMS dosage, temperature, pH and salinity on algal removal efficiency. Algal cells could be rapidly removed by NMS (1-2g/L) through adsorption and physical interaction. The algal destruction process was enhanced under the following reaction conditions: temperature>25°C, salinity>30 ppt and pH value<7.5. The reusability of magnetic recycled NMS and effect of light irradiation on algal cell removal were also determined. NMS exhibited excellent stability after repeated algal cell removal, and the efficiency was further enhanced by light illumination. The current study suggested that using NMS to control C. marina blooms could be a novel promising strategy, which is cost-effective, stable, and easy for recycling., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Developing a strain with high docosahexaenoic acid (DHA) yield and stable fermenting-performance is an imperative way to improve DHA production using Aurantiochytrium sp., a microorganism with two fatty acid synthesis pathways: polyketide synthase (PKS) pathway and Type I fatty acid synthase (FAS) pathway. This study investigated the growth and metabolism response of Aurantiochytrium sp. CGMCC 6208 to two inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan), and proposed a method of screening high DHA yield Aurantiochytrium sp. strains with heavy ion mutagenesis and pre-selection by synergistic usage of cold stress (4°C) and FAS inhibitors (triclosan and isoniazid). Results showed that (1) isoniazid and triclosan have positive effects on improving DHA level of cells; (2) mutants from irradiation dosage of 120Gy yielded more DHA compared with cells from 40Gy, 80Gy treatment and wild type; (3) DHA contents of mutants pre-selected by inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan)at 4°C, were significantly higher than that of wild type; (4) compared to the wild type, the DHA productivity and yield of a mutant (T-99) obtained from Aurantiochytrium sp. CGMCC 6208 by the proposed method increased by 50% from 0.18 to 0.27g/Lh and 30% from 21 to 27g/L, respectively. In conclusion, this study developed a feasible method to screen Aurantiochytrium sp. with high DHA yield by a combination of heavy-ion mutagenesis and mutant-preselection by FAS inhibitors and cold stress., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The effects of five natural products from Chinese herbs including evodiamine, curcumin, 4-methoxysalicylaldehyde, esculin hydrate, and gramine on the growth of Chattonella marina, one of the most noxious red tide algae, were observed. Among them, gramine exhibited the highest inhibitory rate with LC50, 96h of 0.51 mg/l. After exposure to gramine, the activities of superoxide dismutase (SOD) and catalase (CAT), and content of malondialdehyde (MDA) increased in C. marina, suggesting that gramine could induce microalgae oxidative stress. In addition, chlorophyll a and the maximum quantum yield of photosynthesis (Fv/Fm) decreased following exposure to gramine, indicating the inhibition of photosynthesis activity in the microalgae. Combined with the fast inhibition against the algal cells and environmentally friendly character of gramine, we proposed that gramine might be a potential algaecide against marine harmful algae and that the oxidative damage and photosynthesis inhibition might be responsible for the toxicity of gramine on harmful algae.

Heterotrophic microalgae, Aurantiochytrium sp. KRS101 had a large amount of lipid (56.8% total lipids). The cells in the culture medium were easily ruptured due to thin cell wall of Aurantiochytrium sp., which facilitated in-situ fatty acid methyl esters (FAMEs) production directly from biomass. The harvested biomass had a high content of free fatty acids (FFAs), which was advantageous for glycerol-free FAMEs production. FAMEs were directly produced from Aurantiochytrium sp. KRS101 biomass (48.4% saponifiable lipids) using Novozyme 435-catalyzed in-situ esterification in dimethyl carbonate (DMC). DMC was used as a lipid extraction reagent, acyl acceptor and reaction medium. A 433.09mg FAMEs/g biomass was obtained with 89.5% conversion under the optimal condition: DMC to biomass ratio of 5:1 (v/w) and enzyme to biomass ratio of 30% (w/w) at 50°C for 12h. Glycerol could not be detected in the produced FAMEs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Adaptive laboratory evolution (ALE) is an effective method in changing the strain characteristics. Here, ALE with high oxygen as a selection pressure was applied to improve the production capacity of Schizochytrium sp. Results showed that cell dry weight (CDW) of endpoint strain was 32.4% higher than that of starting strain. But slight lipid accumulation impairment was observed. These major performance changes were accompanied with enhanced isocitrate dehydrogenase enzyme activity and reduced ATP:citrate lyase enzyme activity. And a serious decrease of 62.6% in SDHA 140rpm→170rpm was observed in the endpoint strain. To further study the docosahexaenoic acid (DHA) production ability of evolved strain, fed-batch strategy was applied and 84.34g/L of cell dry weight and 26.40g/L of DHA yield were observed. In addition, endpoint strain produced greatly less squalene than starting strain. This work demonstrated that ALE may be a promising tool in modifying microalga strains., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Objectives: To biochemically characterize synthetic peptides to control harmful algal blooms (HABs) that cause red tides in marine water ecosystems., Results: We present an analysis of several short synthetic peptides and their efficacy as algicidal agents. By altering the amino acid composition of the peptides we addressed the mode of algicidal action and determine the optimal balance of cationic and hydrophobic content for killing. In a controlled setting, these synthetic peptides disrupted both plasma and chloroplast membranes of several species known to result in HABs. This disruption was a direct result of the hydrophobic and cationic content of the peptide. Furthermore, by using an anti-HAB bioassay in scallops, we determined that these peptides were algicidal without being cytotoxic to other marine organisms., Conclusions: These synthetic peptides may prove promising for general marine ecosystem remediation where HABs have become widespread and resulted in serious economic loss.

Aquatic organisms are often exposed to dramatic changes in salinity in the environment. Despite decades of research, many questions related to molecular and physiological mechanisms mediating sensing and adaptation to salinity stress remain unanswered. Here, responses of Vaucheria erythrospora, a turgor-regulating xanthophycean alga from an estuarine habitat, have been investigated. The role of ion uptake in turgor regulation was studied using a single cell pressure probe, microelectrode ion flux estimation (MIFE) technique and membrane potential (Em ) measurements. Turgor recovery was inhibited by Gd(3+) , tetraethylammonium chloride (TEA), verapamil and orthovanadate. A NaCl-induced shock rapidly depolarized the plasma membrane while an isotonic sorbitol treatment hyperpolarized it. Turgor recovery was critically dependent on the presence of Na(+) but not K(+) and Cl(-) in the incubation media. Na(+) uptake was strongly decreased by amiloride and changes in net Na(+) and H(+) fluxes were oppositely directed. This suggests active uptake of Na(+) in V. erythrospora mediated by an antiport Na(+) /H(+) system, functioning in the direction opposite to that of the SOS1 exchanger in higher plants. The alga also retains K(+) efficiently when exposed to high NaCl concentrations. Overall, this study provides insights into mechanisms enabling V. erythrospora to regulate turgor via ion movements during hyperosmotic stress., (© 2014 John Wiley & Sons Ltd.)

Increasing demand for renewable fuels has researchers investigating the feasibility of alternative feedstocks, such as microalgae. Inherent advantages include high potential yield, use of non-arable land and integration with waste streams. The nutrient requirements of a large-scale microalgae production system will require the coupling of cultivation systems with industrial waste resources, such as carbon dioxide from flue gas and nutrients from wastewater. Inorganic contaminants present in these wastes can potentially lead to bioaccumulation in microalgal biomass negatively impact productivity and limiting end use. This study focuses on the experimental evaluation of the impact and the fate of 14 inorganic contaminants (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Sb, Se, Sn, V and Zn) on Nannochloropsis salina growth. Microalgae were cultivated in photobioreactors illuminated at 984 µmol m(-2) sec(-1) and maintained at pH 7 in a growth media polluted with inorganic contaminants at levels expected based on the composition found in commercial coal flue gas systems. Contaminants present in the biomass and the medium at the end of a 7 day growth period were analytically quantified through cold vapor atomic absorption spectrometry for Hg and through inductively coupled plasma mass spectrometry for As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Sb, Se, Sn, V and Zn. Results show N. salina is a sensitive strain to the multi-metal environment with a statistical decrease in biomass yieldwith the introduction of these contaminants. The techniques presented here are adequate for quantifying algal growth and determining the fate of inorganic contaminants.

The Cre/loxP site-specific recombination system was applied to Aurantiochytrium limacinum to obtain a transformant without the antibiotic resistance marker gene. First, the enhanced green fluorescent protein gene (egfp) and chloramphenicol resistance gene (Cmr), along with the two loxP loci, were integrated into the genome of A. limacinum OUC88 using 18S rDNA sequences as the homologous recombination sites. Then plasmid pSH65, containing a zeocin resistance gene (Bler) was transferred into A. limacinum OUC&#95;CG. After induction with galactose, repeated passage in culture and PCR-based assessment, the pSH65 plasmid was lost and A. limacinum OUC&#95;EG host was shown to no longer have resistance to 100 mg chloramphenicol/L or 5 mg zeocin/L. Through southern blotting and fluorescence detection, egfp was found to be integrated into the genome of A. limacinum OUC&#95;EG, and EGFP was successfully expressed in the cells. The successful application of the Cre/loxP system demonstrates an experimental basis for genetic modification of A. limacinum so as to obtain transformed strains with no antibiotic resistance marker genes.

Biodiesel produced from microalgal lipids is being considered as a potential source of renewable energy. However, a number of hurdles will have to be overcome if such a process is to become practical. One important factor is the volumetric production of biomass and lipid that can be achieved. The marine alga Nannochloropsis gaditana is under study since it is known to be highly oleaginous and has a number of other attractive properties. Factors that might be important in biomass and lipid production by this alga are light intensity, inoculum size and CO2. Here we have carried out for the first time a RSM-DOE study of the influence of these important culture variables and define conditions that maximize biomass production, lipid content (BODIPY® fluorescence) and total lipid production. Moreover, flow cytometry allowed the examination on a cellular level of changes that occur in cellular populations as they age and accumulate lipids., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A new isolated Schizochytrium sp. LU310 from the mangrove forest of Wenzhou, China, was found as a high producing microalga of docosahexaenoic acid (DHA). In this study, the significant improvements for DHA fermentation by the batch mode in the baffled flasks (i.e. higher oxygen supply) were achieved. By applied the nitrogen-feeding strategy in 1000 mL baffled flasks, the biomass, DHA concentration and DHA productivity were increased by 110.4%, 117.9% and 110.4%, respectively. Moreover, DHA concentration of 21.06 g/L was obtained by feeding 15 g/L of glucose intermittently, which was an increase of 41.25% over that of the batch mode. Finally, an innovative strategy was carried out by intermittent feeding carbon and simultaneously feeding nitrogen. The maximum DHA concentration and DHA productivity in the fed-batch cultivation reached to 24.74 g/L and 241.5 mg/L/h, respectively., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objectives: To investigate the roles of fatty acid biosynthesis in carotenogenesis in the high-lipid accumulating aerial microalga Vischeria helvetica KGU-Y001, we cultured algal cells with fatty acid biosynthesis inhibitors., Results: Under nitrogen-deficient, high-light (200 µmol photons m(-2) s(-1)) conditions, the alga accumulated 6.2 mg carotenoids g(-1) dry weight cells (DWC) after 1 week of culture. The total fatty acid content increased gradually, and reached 290 mg g(-1) DWC after 9 weeks. When algal cells were cultured with a fatty acid biosynthesis inhibitor (molinate) under nitrogen-deficient, high-light conditions for 1 week, carotenoid accumulation was synergistically increased to 2.4 times that in algal cells cultured without the inhibitor in nitrogen-deficient, low-light conditions (40 µmol photons m(-2) s(-1)). The synergistic induction of carotenogenesis was suppressed by an inhibitor of c-jun N-terminal kinase, a mitogen-activated protein kinase-like protein., Conclusion: In a commercial context, carotenoid production could be increased by using fatty acid biosynthesis inhibitors to redirect metabolic flux to carotenoid biosynthesis instead of fatty acid synthesis.

In this work, a newly isolated marine thraustochytrid strain, Schizochytrium sp. DT3, was used for omega-3 fatty acid production by growing on lignocellulose biomass obtained from local hemp hurd (Cannabis sativa) biomass. Prior to enzymatic hydrolysis, hemp was pretreated with sodium hydroxide to open the biomass structure for the production of sugar hydrolysate. The thraustochytrid strain was able to grow on the sugar hydrolysate and accumulated polyunsaturated fatty acids (PUFAs). At the lowest carbon concentration of 2%, the PUFAs productivity was 71% in glucose and 59% in the sugars hydrolysate, as a percentage of total fatty acids. Saturated fatty acids (SFAs) levels were highest at about 49% of TFA using 6% glucose as the carbon source. SFAs of 41% were produced using 2% of SH. This study demonstrates that SH produced from lignocellulose biomass is a potentially useful carbon source for the production of omega-3 fatty acids in thraustochytrids, as demonstrated using the new strain, Schizochytrium sp. DT3., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Ballast water exotic discharge has been identified as a leading vector for marine species invasion. Here immobilized TiO2 photocatalysis is introduced to improve the performance of intense pulsed light. For intense pulsed light/TiO2 photocatalysis, a typical inactivation of 99.89±0.46% can be achieved under treatment condition of 1.78 L min(-1) flow rate, 300 V pulse peak voltage, 15 Hz pulse frequency and 5 ms pulse width. Moreover, within tested 220-260 V peak voltage, 18.37-40.51% elevation in inactivation is observed in comparison with intense pulsed light treatment alone. The rough energy consumption of the tested intense pulsed light/TiO2 treatment system is about 1.51-2.51 times higher than that of the typical commercial UV ballast water treatment system. The stability of the photocatalytic reactivity and intactness of loaded TiO2 film is proved within 20-d's test, while local erosion on stainless steel support is observed after 30-d's test. The results indicate that intense pulsed light/TiO2 photocatalysis is likely to be a competitive ballast water treatment technique, while further measures is needed to reduce the energy consumption and ensure the performance of TiO2 film in a long run., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The oleaginous microalga Nannochloropsis sp. has been spotlighted as a promising candidate in genetic engineering research for biodiesel production. However, one of the major bottlenecks in the genetic manipulation against Nannochloropsis sp. is low transformation efficiency. Based on the idea that they grow rapidly in broth culture, the effect of conditioned medium on colonization and transformation efficiency of Nannochloropsis salina was investigated. Cells grown on agar plates with 20-40% conditioned medium produced colonies that were approximately 2.3-fold larger than cells grown without conditioned medium. More importantly, the transformation efficiency was about 2-fold greater on plates with 30% conditioned medium relative to those without conditioned medium. In addition, FAME productivity in liquid cultures with 100% conditioned medium increased up to 20% compared with cultures of control medium. These results suggest that conditioned medium can be applied for efficient transformation and cost-effective cultivation of N. salina for biodiesel production., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Glucose and glycerol are useful carbon sources for the cultivation of Aurantiochytrium limacinum SR21. Glucose facilitates rapid growth and lipid synthesis, and glycerol promotes the accumulation of docosahexaenoic acid (DHA) in A. limacinum SR21. To improve the DHA productivity of A. limacinum SR21, shake flask and fed-batch cultures were performed using glucose and glycerol as mixed carbon sources (MCSs). Along with optimization of the MCSs, the best DHA yield and productivity (32.36 g/L and 337.1 mg/L/h) were obtained via fed-batch fermentation with maintenance of a constant air supply. The DHA productivity was 15.24% higher than that obtained using glucose as single carbon source (SCS). This study presents a highly efficient and economic strategy for the production of DHA by A. limacinum SR21., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

CO2-expanded ethanol (CXE) was used to extract DHA-containing lipids from Schizochytrium sp. with a 35.7 wt% lipid content of dry biomass in a continuous mode. The effects of operation variables such as temperature, pressure, ethanol flow rate and CO2 flow rate on extraction performance were investigated. Based on a 2(4)-central composite design and response surface methodology, the optimal operating conditions were determined to be a pressure of 6.9 MPa, a temperature of 313 K, an ethanol flow rate of 1 mL/min and a CO2 flow rate of 6.0 mL/min, providing an extracted lipid yield of 87 wt% over an extraction period of 30 min. Not only the lipid yield obtained using CXE was observed to be significantly greater than those using ethanol and pressurized ethanol as the solvents, but also a lower amount of ethanol and less time were required to achieve the same extraction yield by using CXE., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This work reports for the first time the detailed impacts of dual nitrogen (N) and phosphorus (P) stress on growth dynamics and biochemical composition in the Eustigmatophyte Nannochloropsis sp. P-stress concurrent with N-stress had subtle effects on culture bulk biochemical composition, but negatively influenced biomass productivity. However, the N:P supply ratio can be raised to at least 32:1 without compromising productivity (yielding a maximum lipid content of 52% of dry weight and volumetric lipid concentration of 233 mg L(-1)). The maximum biomass and lipid yields per unit of cell-P were 1.2 kg DW (gP)(-1) and 0.54 kg lipid (gP)(-1). The P concentration of many common media is thus in surplus for optimal Nannochloropsis sp. biomass and lipid production, offering potential for significant savings in P usage and improving the sustainability of algal cultivation., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Four nutrient limitation cultures, namely monosodium glutamate (MSG-L), phosphate (P-L), ammonium sulfate (NH4(+)-L) and double (D-L, MSG and P limitation) limited, were designed to study how cell growth and biochemical components of Schizochytrium sp. were affected by nutrient limitations. All limited conditions caused decrease in biomass especially MSG-L and D-L conditions. MSG-L condition attained the highest lipid yield of 30.73 g/l but the lowest protein content. P-L condition shortened the fermentation time and obtained the highest DHA productivity of 291 mg/lh. D-L condition was the most cost-effective fermentation condition which gained the highest input-output ratio. NH4(+)-L condition got the highest squalene and DHA content in lipids. Meanwhile, nitrogen limited conditions promoted the accumulation of neutral lipids. All limited conditions benefit the PUFAs accumulation in the neutral lipids. In addition, the existence of NH4(+) or the absence of MSG and phosphate reduced the unsaponifiable matters content in lipid., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The proteins encoded by psaA and psaB form a heterodimer, an essential compound of photosystem; while the protein encoded by psbC binds with chlorophyll a in photosystem II, serving as antennae in photosystem. Here we report that a heterocyclic brominated flame retardant, tris(2,3-dibromopropyl) isocyanurate (TBC), inhibited the expression of psaA and psbC, then leads to the decrease of Nannochloropsis sp.'s growth biomass. TBC exposure inhibited the expression of psaA and psbC at 10, 100ng/mL slightly and 1000ng/mL significantly. In addition, TBC was found to slow down the growth of Nannochloropsis sp. at concentrations ranging from 10 to 1000ng/mL. These results indicated that TBC influenced both photosynthesis and growth performance of Nannochloropsis sp., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The Fukushima 1 Nuclear Power Plant accident in March 2011 released an enormously high level of radionuclides into the environment, a total estimation of 6.3 × 10¹⁷ Bq represented by mainly radioactive Cs, Sr, and I. Because these radionuclides are biophilic, an urgent risk has arisen due to biological intake and subsequent food web contamination in the ecosystem. Thus, urgent elimination of radionuclides from the environment is necessary to prevent substantial radiopollution of organisms. In this study, we selected microalgae and aquatic plants that can efficiently eliminate these radionuclides from the environment. The ability of aquatic plants and algae was assessed by determining the elimination rate of radioactive Cs, Sr and I from culture medium and the accumulation capacity of radionuclides into single cells or whole bodies. Among 188 strains examined from microalgae, aquatic plants and unidentified algal species, we identified six, three and eight strains that can accumulate high levels of radioactive Cs, Sr and I from the medium, respectively. Notably, a novel eustigmatophycean unicellular algal strain, nak 9, showed the highest ability to eliminate radioactive Cs from the medium by cellular accumulation. Our results provide an important strategy for decreasing radiopollution in Fukushima area.

The occurrence of harmful algal blooms has been increasing significantly around the world. In order to ensure the safety of drinking water, procedures to screen potential materials as effective algicides are needed, and predictive methods which save both the labor and time compared with traditional experimental approaches, are particularly desirable. In this study, data from previous studies on the algal-growth inhibitory action of two kinds of compounds, namely, the action of fatty acids and thiazolidinediones on the harmful algae Heterosigma akashiwo and Chattonella marina, were modeled using multiple linear regression (MLR) based on quantitative structure-activity relationships (QSAR). The models were shown to have highly predictive ability and stability, and provided insight into the inhibitory mechanisms of congeneric compounds. The main descriptors in the fatty-acid models were the Connolly accessible area and the number of rotatable bonds, illustrating that molecular surface area and shape are important in their algicidal actions. In the thiazolidinedione models, the critical volume, octanol-water partition coefficient (LogP), and Connolly solvent-excluded volume were found to be significant, indicating that hydrophobicity, substituent group size, and mode of action are mechanistically important. Our results showed the algicidal activity of a series of compounds on different algae could be modeled, and each model is efficacious for compounds that fall into the application domain of the QSAR model. This work demonstrates how reliable predictions of the algicidal activity of novel compounds and explanations of their inhibitory mechanisms can be obtained.

A cell's growth phase could affect its susceptibility to a biocide in microbial control. This study examines the growth phase dependent susceptibility of a brown tide bloom alga Aureococcus anophagefferens to microbial biocide hydrogen peroxide (H2O2). Test cultures of A. anophagefferens cells in exponential and stationary growth phase and similar initial cell density (1.6×10(6) cells mL(-1)) were exposed to 0.4-1.6 mg L(-1) H2O2. Changes in algal growth (in vivo fluorescence, total chlorophyll a, and cell density), cell physiology (maximum quantum yield of photosystem II, and total intracellular non-protein thiols), and H2O2 decomposition were quantified. Results show that the stationary phase cells are more susceptible to H2O2 than the exponential phase cells, and this is attributed to the weaker ROS (reactive oxygen species) scavenging system and consequently greater cell damage in stationary phase cells. The stationary phase cells potentially require 30-40% less H2O2 to reach 90% removal within 12 h of treatment as compared to the exponential phase cells. The results have practical implications in brown tide bloom control with respect to the timing and the dosage of H2O2 application., (Copyright © 2013 Elsevier B.V. All rights reserved.)

The potential use of Synechocystis sp. PCC6803 (Synechocystis sp.) for lipid production using artificial seawater (ASW) medium supplemented with anaerobic digestion effluent (ADE) was investigated and compared to marine microalgae, Nannochloropsis salina (N. salina). Synechocystis sp. showed growth rates 83% and 20% higher than N. salina at 3% and 6% ADE loading ratios, respectively, achieving the highest biomass productivity of 212 mg L(-1) d(-1) in semi-continuous cultivation. The rapid growth of Synechocystis sp. was offset by its low lipid content, resulting in lipid productivities 7-28% lower than N. salina. The lipid productivity of Synechocystis sp. may be further improved by decreasing the harvesting interval during semi-continuous cultivation. Fatty acid analysis showed that lipids extracted from Synechocystis sp. contained higher palmitic acid (60.3±2.0%) and linoleic acid (20.0±1.6%), and had a higher cetane number and oxidative stability than those from N. salina., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The trace element selenium (Se) is required for the biosynthesis of selenocysteine (Sec), the 21st amino acid in the genetic code, but its role in the ecology of harmful algal blooms (HABs) is unknown. Here, we examined the role of Se in the biology and ecology of the harmful pelagophyte, Aureococcus anophagefferens, through cell culture, genomic analyses, and ecosystem studies. This organism has the largest and the most diverse selenoproteome identified to date that consists of at least 59 selenoproteins, including known eukaryotic selenoproteins, selenoproteins previously only detected in bacteria, and novel selenoproteins. The A. anophagefferens selenoproteome was dominated by the thioredoxin fold proteins and oxidoreductase functions were assigned to the majority of detected selenoproteins. Insertion of Sec in these proteins was supported by a unique Sec insertion sequence. Se was required for the growth of A. anophagefferens as cultures grew maximally at nanomolar Se concentrations. In a coastal ecosystem, dissolved Se concentrations were elevated before and after A. anophagefferens blooms, but were reduced by >95% during the peak of blooms to 0.05 nM. Consistent with this pattern, enrichment of seawater with selenite before and after a bloom did not affect the growth of A. anophagefferens, but enrichment during the peak of the bloom significantly increased population growth rates. These findings demonstrate that Se inventories, which can be anthropogenically enriched, can support proliferation of HABs, such as A. anophagefferens through its synthesis of a large arsenal of Se-dependent oxidoreductases that fine-tune cellular redox homeostasis.

An efficient magnetic separation technology using Fe(3)O(4) nanoparticles was developed for harvesting marine microalgae Nannochloropsis maritima from culture broth. Recovery capacity of these nanoparticles was affected by microalgal growth phase and reached the peak value when the microalgal growth reached its maximal biomass after 18 days. The recovery efficiency of microalgal cells from the culture medium reached more than 95% at the particle dosage of 120 mg/L within 4 min. Electrostatic attraction at acidic pH and cell aggregation under neutral and alkaline conditions was beneficial for harvesting the algal cells. Higher operation temperature resulted in higher adsorption capacity of these nanoparticles for microalgawl cells. Reuse of the culture medium obtained from magnetic separation gave similar biomass production in comparison with that from centrifugation separation after 5 recycles. Together with these results provide a great potential in high-efficient and economical harvesting of tiny marine microalgae using magnetic separation technology in practice., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Protozoans of the family Opalinidae are intestinal commensals in amphibians. To test the hypothesis that these organisms are susceptible to the antiprotozoal antibiotic metronidazole, we randomly assigned 60 juvenile Woodhouse's toads ( Bufo woodhousii ) to receive a single oral dose of metronidazole or water. In pilot trials, the prevalence of opalinids in untreated members of this population was over 70%. One-third of the study population was dissected at each of 3 time points: 18 hr, 1 wk, and 2 wk post-treatment. An examiner blinded to the toad's treatment history determined the presence or absence of opalinids using a dissecting microscope. Opalinids were found in 3/10 toads in the treatment group and 9/10 in the control group after 18 hr (P < 0.02), in none of the treatment group and 8/10 in the control group after 1 wk (P < 0.001), and in none of the treatment group and 10/10 in the control group after 2 wk (P < 0.0001). These results suggest that a single-dose of metronidazole quickly and reliably clears opalinids from juvenile Woodhouse's toads with no evidence of short-term recurrence. The treatment was well tolerated, with no apparent morbidity and no mortality in either group. Future exploration of opalinid-related host fitness consequences may be facilitated by this simple method of developing a protozoan-free host population.

The biology of Aurantiochytrium sp. KRS101 was thoroughly investigated to enhance its production of biodiesel and docosahexaenoic acid (DHA). Nutrients and salinity were optimized to prevent biomass loss due to cell rupture. Calculation of yield coefficients showed that nitrogen was mostly responsible for the early stage of cell growth or division, whereas carbon was necessary for the entire process of cell development, particularly cell enlargement during late stages. Using these distinctive yield coefficients, a modified fed-batch cultivation method was designed, resulting in increases in palmitic acid (PA) and DHA production of up to 137% and 29%, respectively. This modified fed-batch cultivation method, using appropriate supplies of nitrogen and carbon, may improve the yields of PA and DHA, thus expanding the biotechnological applications of Aurantiochytrium sp. KRS101., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Nanoecotoxicology as a sub-discipline of ecotoxicology aims to identify and predict effects elicited on ecosystems by nano-sized materials (NM). Two key groups of model organisms in this context are algae and fish. In this chapter, we present considerations for testing NM with respect to their impact on unicellular algae and cell lines derived from various organs of fish.Based on currently available literature on NM effects in unicellular algae and fish cell lines, and our own experience, we provide guidance on test design, including principle test considerations, materials, NM presentation to cells, exposure, bioavailability, and effect assessment. Assessment needs to be based on a meaningful choice of exposure scenario(s) related to the research question. As a first step, one needs to address whether effects of NMs are to be investigated under environmentally relevant or probable conditions, which may include processes such as agglomeration, or whether NM effects from mono-dispersed particles are of interest, which may require special steps to ensure stable NM suspension. Moreover, whether effects on cells are to be studied in the short- or long-term is important with regard to experimental design. Preparation of NM suspensions, which can be done in aqueous media different from the exposure medium, is addressed with regard to energy input, sterility (as required for algae and fish cell exposure) and particle purity.Specified for the two model systems, algae and fish cell lines, availability and choice of culture media are presented and discussed with regard to impact on NM behavior. Light, temperature, and agitation, which are variables during exposure, are discussed. We further provide guidance on the characterization of the NM in the chosen aqueous exposure media regarding size, zeta potential and electrophoretic mobility. The state of NM in exposure media is decisive for their bioavailability and therefore for potential particle effects. Therefore, we present ways of deriving a mass balance and quantitative/qualitative information on the uptake and distribution of NM in cells.As NM have a high surface-to-volume ratio and possess specific physical-chemical properties, which make them prone to interfere with various compounds and certain types of toxicity tests, potential interferences and appropriate controls are introduced. Furthermore, different types of dose metrics, which is still a strongly debated issue in nanotoxicology, are highlighted. We also consider laboratory safety regarding NM handling and disposal.

Marine microalgae represent a potentially valuable feedstock for biofuel production; however, large-scale production is not yet economically viable. Optimisation of productivity and lipid yields is required and the cost of biomass harvesting and dewatering must be significantly reduced. Microalgae produce a wide variety of biologically active metabolites, many of which are involved in inter- and intra-specific interactions (the so-called infochemicals). The majority of infochemicals remain unidentified or uncharacterised. Here, we apply known and candidate (undefined extracts) infochemicals as a potential means to manipulate the growth and lipid content of Nannochloropsis oculata-a prospective species for biofuel production. Five known infochemicals (β-cyclocitral, trans,trans-2,4-decadienal, hydrogen peroxide, norharman and tryptamine) and crude extracts prepared from Skeletonema marinoi and Dunaliella salina cultures at different growth stages were assayed for impacts on N. oculata over 24 h. The neutral lipid content of N. oculata increased significantly with exposure to three infochemicals (β-cyclocitral, decadienal and norharman); however the effective concentrations affected a significant decrease in growth. Exposure to particular crude extracts significantly increased both growth and neutral lipid levels. In addition, water-soluble extracts of senescent S. marinoi cultures induced a degree of flocculation in the N. oculata. These preliminary results indicate that artificial manipulation of N. oculata cultures by application of algae infochemicals could provide a valuable tool towards achieving economically viable large-scale algae biofuel production.

The occurrence of sixteen pharmaceutically active compounds in influent and effluent wastewater and in primary, secondary and digested sludge in one-year period has been evaluated. Solid-water partition coefficients (Kd) were calculated to evaluate the efficiency of removal of these compounds from wastewater by sorption onto sludge. The ecotoxicological risk to aquatic and terrestrial ecosystems, due to wastewater discharges to the receiving streams and to the application of digested sludge as fertilizer onto soils, was also evaluated. Twelve of the pharmaceuticals were detected in wastewater at mean concentrations from 0.1 to 32 μg/L. All the compounds found in wastewater were also found in sewage sludge, except diclofenac, at mean concentrations from 8.1 to 2206 μg/kg dm. Ibuprofen, salicylic acid, gemfibrozil and caffeine were the compounds at the highest concentrations. LogKd values were between 1.17 (naproxen) and 3.48 (carbamazepine). The highest ecotoxicological risk in effluent wastewater and digested sludge is due to ibuprofen (risk quotient (RQ): 3.2 and 4.4, respectively), 17α-ethinylestradiol (RQ: 12 and 22, respectively) and 17β-estradiol (RQ: 12 and 359, respectively). Ecotoxicological risk after wastewater discharge and sludge disposal is limited to the presence of 17β-estradiol in digested-sludge amended soil (RQ: 2.7)., (Copyright © 2012 Elsevier B.V. All rights reserved.)