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An effective prototype of photobioreactor namely hollow fiber membrane bioreactor is employed to produce algal biomass for biofuel production. The effect of two-stage cultivation system on the biomass productivity and lipid production is studied using freshwater green microalga Tetradesmus obliquus MT188616.1. The hybrid system combines exponential biomass production in hollow fiber membrane photobioreactor (HFMPBr) and a coordinated high-lipid stimulation phase in nitrogen-deprived medium by the shake flask method. This work is proposed to examine the usefulness of HFMPBr module to enhance the microalgal growth rate through the effective mass transfer by standardizing the culture medium re-circulation flow rates (5 - 45 mL min

Recent studies show that triclosan (TCS) exposure causes reduction in pigments, suppression of photosynthesis, and induction of oxidative stress at the physiological level, resulting in morphological alteration and growth inhibition in algae including Raphidocelis subcapitata (R. subcapitata, a freshwater model green alga). However, the underlying molecular mechanisms remain to be elucidated, especially at environmentally relevant concentrations. The present study uncovered the transcriptional profiles and molecular mechanisms of TCS toxicity in R. subcapitata using next-generation sequencing. The algal growth was drastically inhibited following a 7-day exposure at both 75 and 100 μg/L TCS, but not at 5 μg/L (environmentally realistic level). The transcriptomic analysis shows that molecular signaling pathways including porphyrin and chlorophyll metabolism, photosynthesis – antenna proteins, and photosynthesis were suppressed in all three TCS treatments, and the perturbations of these signaling pathways were exacerbated with increased TCS exposure concentrations. Additionally, signaling of replication-coupled DNA repair was only activated in 100 μg/L TCS treatment. These results indicate that photosynthesis systems were sensitive targets of TCS toxicity in R. subcapitata, which is distinct from the inhibition of lipid synthesis by TCS in bacteria. This study provides novel knowledge on molecular mechanisms of TCS toxicity in R. subcapitata.

Ciprofloxacin (CIP) is an antimicrobial “pseudo-persistent” in aquatic ecosystems. Once dispersed in the water compartments, it can also affect the microalgae. Thus, the evaluation of its long-term ecotoxicological effects is necessary. CIP interactions with other pharmaceuticals are not well known. In this study, we investigated the toxic effects of CIP alone and combined with caffeine (CAF), using the modified Gompertz model parameters and the chlorophyll-a production of the microalga Raphidocelis subcapitata as endpoints, throughout a 16-day exposure assay. The exposure to CIP alone led to significant reductions of the growth rate and the cell density of the microalgae compared to control groups. The combination with CAF lowered the adverse effects of CIP to R. subcapitata. However, as the toxicity is dynamic, our results indicated that the toxic effects in respect to the studied endpoints changed throughout the exposure period, reinforcing the need for longer-term ecotoxicity assessments.

We isolated fifty-two strains from the marine aquaculture ponds in Malaysia that were evaluated for their lipid production and ammonium tolerance and four isolates were selected as new ammonium tolerant microalgae with high-lipid production: TRG10-p102 Oocystis heteromucosa (Chlorophyceae); TRG10-p103 and TRG10-p105 Thalassiosira weissflogii (Bacillariophyceae); and TRG10-p201 Amphora coffeiformis (Bacillariophyceae). Eicosapentenoic acid (EPA) in three diatom strain was between 2.6 and 18.6 % of total fatty acids, which were higher than in O. heteromucosa. Only A. coffeiformi possessed arachidonic acid. Oocystis heteromucosa naturally grew at high ammonium concentrations (1.4–10 mM), whereas the growth of the other strains, T. weissflogii and A. coffeiformi, were visibly inhibited at high ammonium concentrations (>1.4 mM-NH4). However, two strains of T. weissflogii were able to grow at up to 10 mM-NH4 by gradually acclimating to higher ammonium concentrations. The ammonium tolerant strains, especially T. weissflogii which have high EPA contents, were identified as a valuable candidate for biomass production utilizing NH4-N media, such as ammonium-rich wastewater.

The present study evaluated the effects of natural astaxanthin (ASTA) from Haematococcus pluvialis on the antioxidant capacity, lipid metabolism, and ASTA accumulation in the egg yolk of laying hens. Hy-Line Brown layers (n = 288, 50 wk old) were randomly assigned to 1 of 4 dietary treatment groups. Each group had 6 replicates of 12 hens each. All birds were given a corn-soybean meal–based diet containing 0, 25, 50, or 100 mg/kg ASTA for 6 wk. The results showed that the total antioxidant capacity, superoxide dismutase level, and glutathione peroxidase level in the plasma, livers, and egg yolks were significantly increased in the ASTA groups compared with those of the control group (P

Astaxanthin is a highly valuable derivative of β-carotene, as well as an important raw material for pharmacy and healthy industries. Dunaliella salina is a halotolerant green alga that contains manifold nutriments and possesses a remarkable ability to adapt environment. The alga is also able to effectively accumulate β-carotene up to 10% of its dry weight. However, D. salina is lack of the enzymes for astaxanthin synthesis so that the accumulated β-carotene cannot be converted to astaxanthin in the alga. The designed experiment was that the exogenous key enzyme genes for astaxanthin synthesis, β-carotene ketolase and β-carotene hydroxylase cloned from Haematococcus pluvialis (Chlorophyceae), were co-transformed into D. salina by bombardment deliver system. The observation of laser scanning confocal microscope and analysis of PCR amplification confirmed the efficient expression of the transformed genes. Compared with the wild type strain, the transformants showed stronger high irradiation adaptability with higher cell survival rate, light saturation point, maximum relative electron transport rate and lower ROS level. The results of biochemical analysis indicated that the activities of antioxidant enzymes showed no significant difference between the two groups. At the transcriptional level, four carotenogenic genes (PSY, PDS, ZDS, LCYB) expressed 2~4 fold higher in the transformants than in the wild type. The tested transformants treated under high irradiation intensity were able to synthesize astaxanthin and the amount of astaxanthin was 5.56 μg/g dry weight measured by HPLC. This study may provide fundamental data for metabolic engineering to synthesize astaxanthin in D. salina.

The present study was focused to evaluate the phytoplankton biodiversity in two freshwater perennial lakes of Sulur and Ukkadam of Coimbatore District, Tamil Nadu, India. The study period was two years from March 2012 to February 2014. Results revealed that totally 50 species of phytoplankton include 14 Cyanophyceae, 19 Chlorophyceae, 14 Bacillariophyceae, and 3 Euglenophyceae were recorded from the Sulur lake and 36 species of phytoplankton comprising 9 species of Cyanophyceae, 15 species of Chlorophyceae, 10 species of Bacillariophyceae and 2 species of Euglenophyceae were noticed in Ukkadam lake during the study period. The recorded phytoplankton density was ranged between 22,380 and 80,590 cells/l in Sulur lake and 29,350 and 1,16,255 cells/l in Ukkadam lake. Thus, the present study overall dominance of phytoplankton group was Chlorophyceae followed by Cyanophyceae, Bacillariophyceae, and Euglenophyceae. The phytoplankton population was recorded maximum in Ukkadam lake when compared to Sulur lake. Hence, the study revealed that both Ukkadam and Sulur lakes possess an appreciable quantum of phytoplankton and it can be used for fish culture.

The challenges associated with effective cell wall disruption remain an important bottleneck that has restricted efforts to extract astaxanthin from Haematococcus pluvialis. Here, available transcriptomic data and an Agrobacterium tumefaciens-mediated transformation system were used to establish an H. pluvialis strain in which the key cell wall formation-related enzyme α-1,6-mannosyltransferase (HpOCH1) was downregulated in an effort to thin cell walls and thereby simplify the astaxanthin extraction process. The cell wall remodeling activity observed in these HpOch1 knockdown H. pluvialis cells resulted in dramatic reductions in the mannan organization and protective ability of the established cell walls. The cell fragmentation rate increased by 58% in HpOch1

Microalgal-based processes offer promise for addressing two sustainability challenges: recovering phosphorus (P) from wastewater and producing biofuel feedstock. This study investigated the role of phosphorus supply on microalgal growth, lipid yield, and P distribution for Scenedesmus during nitrogen starvation. Extracellular polymeric substances and intracellular polymeric substances were the most important pools for inorganic phosphorus (IP) and organic phosphorus (OP), respectively. The main P pool for microalgae with low phosphorus supply was EPS, which accounted for 57 % of the total biomass phosphorus; while under high P concentrations, 79 % of the phosphorus was stored in IPS. A high concentration of orthophosphate stimulated rapid P uptake as IP and promoted the transformation of IP to OP associating with biomass synthesis. The highest P content of microalgal biomass was 6.5 % of dry weight when the phosphorus concentration in medium was 113 mg/L, and the OP content was 4.9 % of dry weight. High phosphate-P enhanced the biomass's lipid content by 60 %, and the distribution of fatty acid methyl esters was not altered by P concentrations. Collectively, high phosphate-P availability could promote microalgal biomass synthesis, lipid production and P accumulation.

The toxicity of microplastics (MPs) to marine microalgae has raised much concern. However, research at metabolic level is quite limited. In this study, the physiological and metabolic effects of polystyrene (PS) and aged polystyrene (A-PS) MPs on Dunaliella salina were investigated. Both PS and A-PS inhibited the growth of microalgae, but promoted the pigment synthesis in algal cells. The oxidative stress analysis indicated that PS and A-PS induced high production of reactive oxygen species (ROS), and caused oxidative damage to algal cells. Particularly, the highest ROS level in PS and A-PS groups were 1.70- and 2.24-fold of that in the control group, respectively. Untargeted metabolomics analysis indicated that PS and A-PS significantly increased the differential metabolites. Compared with the control group, the significant upregulation of glycerophospholipids metabolites illustrated that severe membrane lipid peroxidation occurred in the MPs groups. Metabolic pathways analysis showed that PS and A-PS perturbed the amino acid-related metabolic pathways. In particular, the amino acid biosynthesis and ATP-binding cassette (ABC) transporter pathways were significantly upregulated, thus promoting nitrogen storage and transmembrane transport in Dunaliella salina. Transmembrane transport requires a large amount of ATP; as a result, algal cell division is inhibited. In addition, A-PS stimulated more active glutathione metabolism than PS. These results enrich the understanding of the toxicity of PS MPs to microalgae at the metabolic level, and are helpful for further assessing the ecological impacts of MPs on microalgae.

In the present study, exogenous myo-inositol (MI) was applied to induce natural astaxanthin and biolipid accumulation in Haematococcus pluvialis. Under 200 μM MI, algal cells exhibited 62.11 % and 34.67 % increases in astaxanthin and lipid content, respectively, compared to the control. The carotenogenesis and lipogenesis genes were upregulated by induction of MI. Interestingly, MI addition elevated the ethylene (ETH) content and activated antioxidant enzyme-associated gene levels, which could be involved in alleviating oxidative stress. Further data showed that the ETH signal played a positive function in stimulating astaxanthin biosynthesis under MI induction. Supplementation with ethephon plus MI boosted the astaxanthin content to 33.08 ± 0.03 mg g

In the current research, a novel microalgae strain was isolated from Yajiageng Red Rock Beach and identified as Acutodesmus sp. HLGY. To obtain high-efficiency production of lutein from algae, the feasibility of using melatonin (MT) to increase lutein yield of Acutodesmus sp. HLGY was evaluated. Under the 7.5 μM MT treatment, the lutein content and lutein productivity were 17.44 mg g

Mixotrophy of Haematococcus pluvialis is a potential strategy for producing astaxanthin. However, this strategy has not been extensively commercialized because the mixotrophic mechanisms by which H. pluvialis overcomes high light stress are unclear. This study analyzed the biochemical compositions and differential proteomics of mixotrophic H. pluvialis under different light conditions. High light exposure substantially increased astaxanthin, carbohydrate, and fatty acid contents. A total of 119 and 81 proteins were significantly up- and down-regulated after two days of high light exposure. These proteins mainly enriched pathways for photosynthetic metabolism, glyoxylate cycle, and biosynthesis of secondary metabolites. This study proposed a regulatory model through which mixotrophic H. pluvialis copes with high light stress. The model includes pathways for modulating photosynthetic apparatus, increasing astaxanthin accumulation by enhancing photorespiration, pentose phosphate and Embden-Meyerhof-Parna pathways, while thickening the cell wall by malate-oxaloacetate shuttle.

Flocculation has been proved an efficient method for microalgal biomass harvesting, but some coagulant agents may have adverse effects on microalgae growth, making the reuse of the medium unfeasible. In this study, Haematococcus pluvialis was harvested by different flocculants, and the feasibility of the reuse of the culture medium was evaluated. Results suggested that both inorganics, polyaluminum chloride (PA) and ferric chloride (FC), and organics, extracted from Moringa oleifera seed (MSE) and chitosan (CH) resulted in efficient flocculation - flocculation efficiency above 99 %. However, using PA and FC had adverse effects on the astaxanthin recovery from haematocysts - losses of 58.6 and 73.5 %, respectively. Bioflocculants in the reused medium also had higher growth performance than inorganic ones. Furthermore, bioflocculants in reused medium increase the contents of β-carotene, astaxanthin, and linolenic acid. This investigation demonstrated that using MSE and CHI for harvesting H. pluvialis enables the water reusability from a flocculated medium.

Microalgae are photosynthetic microorganisms that stand out from conventional food sources and ingredients due to their high growth rate and adaptability. In addition to being highly sustainable, significant concentrations of proteins, lipids, and pigments accumulate in their cell structures from photosynthesis. Hence, this study sought to evaluate the food potential of Scenedesmus obliquus biomasses obtained from photosynthetic cultures enriched with 3, 5, 10, 15, 20, and 25% carbon dioxide (CO

Microalgae have the highest capability to fix the atmospheric carbon and wastewater-derived nutrients to produce high-value bioproducts including lipids and carotenoids. However, their lower titers and single-product-oriented biomass processing have made the overall process expensive. Hence, increased metabolite titer and processing of the biomass for more than one product are required to ensure the commercial robustness of the algal biorefinery. In this study, a newly isolated algal strain was identified as Bracteacoccus pseudominor BERC09 through phylogenetic analysis based on the 18S rRNA gene sequence. Basic characterization of the strain revealed its promising potential to produce carotenoids and lipids. The lipids and carotenoid biosynthesis pathways of BERC09 were further triggered by manipulating the abiotic factors including nitrogen sources (NaNO

Two cultured chlorophyceae algae and two cyanobacteria-rich phytoplankton were separated into five fractions and characterized by 13C cross polarization/total sideband suppression (CP/TOSS) nuclear magnetic resonance spectroscopy (NMR), X-ray photoelectron spectroscopy (XPS), Rock-Eval and closed pyrolysis analyses. The structures of the acid nonhydrolyzable organic matter (NHOM) fractions were similar to those of poly(methylene)-rich aliphatic algaenan. We found that the cyanobacteria-rich phytoplankton contained aliphatic nonhydrolyzable biopolymers. The NMR and Rock-Eval analyses showed that the isolated NHOM exhibited high oil-prone potentials. Pyrolysis analysis of the field-collected NHOM from Liuhuahu Park in Guangzhou (GP_NHOM) indicated that it exhibited high oil-prone potential and comprised a long, saturated and unbranched hydrocarbon chain (up to C33) structural unit. The maximum oil and gaseous hydrocarbon generation potentials reached 55.9% and 7.2%, respectively and were very close to those derived from 13C CP/TOSS NMR, suggesting that 13C NMR is an effective approach for evaluation of oil and gas production potentials. This investigation is helpful for improving understanding of kerogen formation mechanisms through selective preservation and the generation mechanisms of biocrude oil.

The glycerol-3-phosphate dehydrogenase (GPD) gene family plays a major role in glycerol synthesis and adaptation to abiotic stresses. Few studies on GPD family genes from the halotolerant algae Dunaliella salina are available. In this study, seven DsaGPD genes were identified by mining D. salina sequencing data. Among them, DsaGPD5 contained the canonical NAD+-GPD protein domain, called si-GPD. In comparison, DsaGPD1-4 not only contained the canonical NAD+-GPD domain but also a unique domain, the haloacid dehalogenase (HAD)-like superfamily domain, in their N-terminal region, called bi-GPD. DsaGPD6, 7 contained the FAD+-GPD domain. In the transient expression system, DsaGPD1, 3, 4 were found in the cytosol of Arabidopsis thaliana protoplast, DsaGPD2, 5 in the chloroplast, and DsaGPD6, 7 in the mitochondria. MEME analysis showed that six conserved motifs were present in both si-GPDs and bi-GPDs, whereas seven highly conserved motifs were only present in bi-GPDs. The quantitative real-time PCR results showed significant induction of the DsaGPD genes under abiotic stresses, indicating their tolerance-related role in D. salina. DsaGPD2 and DsaGPD5 may be the osmoregulator form and glyceride form in the chloroplast, respectively. The evolutionary forces acting on si-GPDs and bi-GPDs were different in the same organism: bi-GPDs were under purifying selection, while si-GPDs were mainly under positive selection. Furthermore, evolution of the N_HAD domain and C_GPD domain in bi-GPDs is highly correlated. In summary, this study characterizes DsaGPD gene family members and provides useful information for elucidating the salt tolerance mechanism in D. salina.

Isoprenoid diphosphates are important precursors actively participating in many downstream metabolisms; they are often in modified forms, e.g., protein-coupled or esterified form. Therefore, in vivo level of free isoprenoid diphosphates is quite low, ˜0.07 nmol/g fresh weight in plants. In order to directly measure the isoprenoid diphosphate pool during stress-induced accumulation of astaxanthin in Haematococcus pluvialis, the present study optimized several pretreatment procedures to enrich free isoprenoid diphosphates for high-pressure liquid chromatography with tandem mass spectrometry (HPLC-MS/MS) detection. Specifically, different extraction solvents, e.g., water, methanol, chloroform, and mixture of water, methanol, and chloroform (1:1:1, V/V/V), and solid phase extraction (SPE) columns (OASIS@ WAX and HLB Cartridges) were compared; and gentle decoupling by NaOH or trifluoroacetic acid (TFA) was introduced to release free isoprenoid diphosphates. Results found that solvent mixture of water, methanol and chloroform (1:1:1, V/V/V) showed the highest extraction efficiency (RE) for five isoprenoid diphosphates, ranging from 76.83% to 92.43%; HLB column showed the balanced recoveries ranging from 75.29% to 87.54%; and incubation with low NaOH (˜4.7 mmol/L) at 4 °C significantly increased detectable isoprenoid diphosphates in algal cells, some of which were undetectable or in trace level before NaOH decoupling. The method was applied to H. pluvialis cells under various stresses. Low levels of isoprenoid diphosphates were determined in most of the stresses used, e.g., 0.19 ± 0.09 to 0.98 ± 0.06 mg/g fresh weight (FW) for IPP/DMAPP, 0.35 ± 0.07 mg/g FW for GGPP and undetectable for FPP and GPP; while isoprenoid diphosphates were significantly accumulated in the dark to 3.27 ± 0.05, 0.17 ± 0.09, 1.81 ± 0.16 and 0.58 ± 0.07 mg/g FW for IPP/DMAPP, GPP, FPP and GGPP, respectively. These results implied that isoprenoid diphosphates were exhausted by downstream carotenogenesis under stress. Our work emphasizes NaOH decoupling for exact quantitation of in vivo isoprenoid diphosphates.

A two-step strategy was employed to culture Dunaliella tertiolecta, an oleaginous unicellular green alga, combined by the salt stress and sodium azide intervention, to observe their effects on its lipid accumulation. When the algae cultured at different salt concentrations reached the logarithmic growth phase, sodium azide was added. The results showed that the addition of sodium azide significantly increased the lipid content and had no significant effect on cell biomass. The lipid yield and single cell lipid content under 50 μM sodium azide increased by 10.4% and 21.7%. Under the two-step culture condition, combining of the treatment of 50 μM sodium azide and 2.5 M salt stress, the total lipid productivity and single-cell lipid content were 10% and 70.5% higher than that of the control. It seemed that sodium azide and salinity might have a synergistic effect on the lipid accumulation of D. tertiolecta. It can be concluded that sodium azide is an effective inducer of lipid accumulation in D. tertiolecta, and two-stage cultivation is a feasible way to improve lipid accumulation in microalgae.

Glycerol-3-phosphate (G3P) is the important precursors for triacylglycerol synthesis, while glycerol-3-phosphate dehydrogenase (GPDH) determines the formation of G3P. In this study, two GDPH genes, Dtgdp1 and Dtgdp2 were isolated and identified from Dunaliella tertiolecta. The full-length Dtgdp1 and Dtgdp2 CDS were 2016 bp and 2094 bp, which encoded two putative protein sequences of 671 and 697 amino acids with predicted molecular weights of 73.64 kDa and 76.73 kDa, respectively. DtGDP1 and DtGDP2 both had a close relationship with those of algal and higher plants. DtGDP1 shared two conserved superfamily (A1 and A2) and four signature motifs (I-IV), and the DtGDP2 showed six signature domains (from motif I to VI) and DAO_C conserved family. Our previous work showed that the triethylamine intervention could greatly increase the triacylglycerol content (up to 80%) of D. tertiolecta. This study aims to investigate the effect of triethylamine on GPDH expression. Results showed that, when treated by triethylamine at 100 ppm and 150 ppm, the expression levels of Dtgdp1 and Dtgpd2 were increased to 5.121- and 56.964-fold compared with the control, respectively. Triethylamine seemed to enhance lipid metabolic flow by inducing the expressions of Dtgdp1 and Dtgdp2 to increase the lipid content, which provides a new insight into the desired pathway of lipid synthesis in algae through genetic engineering.

International audience; Nanomechanical and structural characterisations of algal cells are of key importance for understanding their adhesion behaviour at interfaces in the aquatic environment. We examine here the nanomechanical properties and adhesion dynamics of the algal cells during two phases of their growth using complementary surface methods and the mathematical modelling. Mechanical properties of motile cells are hard to assess while keeping cells viable, and studies to date have been limited. Immobilisation of negatively charged cells to a positively charged substrate enables high-resolution AFM imaging and nanomechanical measurements. Cells were stiffer and more hydrophobic in the exponential than in the stationary phase, suggesting molecular modification of the cell envelope during aging. The corresponding properties of algal cells were in agreement with the increase of critical interfacial tensions of adhesion, determined amperometrically. Cells in exponential phase possessed a larger cell volume, in agreement with the large amount of amperometrically measured displaced charge at the interface. Differences in the kinetics of adhesion and spreading of cells at the interface were attributed to their various volumes and nanomechanical properties that varied during cell aging. Our findings contribute to the present body of knowledge on the biophysics of algal cells on a fundamental level.

The effect of waterborne ingredient on ecosystem has been of great interest. In the present study, the evaluation method using algal photosynthesis inhibition assay with dual-channel pulse amplitude modulation (PAM) system was established for a series of water samples to elucidate the potential effect of the total body of organic compounds including natural organic matter (NOM) on aquatic ecosystems. The more sensitive and less time-consuming monitoring method compared with algal growth inhibition assay was suggested, especially considering inorganic and coloring constituents. Algal photosynthesis inhibition activity was detected with high sensitivity for photosystem II (PSII) inhibitors, whereas the IC10 of the other chemicals was over the environmental standard concentration for Chlamydomonas moewusii (Chlorophyceae) and Pheodactylum tricornutum (Diatomea). The photosynthesis inhibition activity of Lake Biwa dissolved organic matter (LBDOM) and fulvic acid (LBFA) was significantly detected at ≥10 times the concentration and >10 mgC L−1, respectively, whereas prominent activity was confirmed for Suwannee River NOM (SRNOM) on the river original concentration (>30 mgC L−1) for both algae. Significant inhibition activity was detected in both algae at least in twice-concentration for water samples from a wastewater treatment pilot plant. There was no great difference in the activity between sewage secondary effluent and its filtrate with ultrafiltration (UF), and physically washing water for the UF membrane.

This study examined the effects of a bacterial community and extracellular polymeric substances (EPS) on Ettlia sp. flocculation. The growth rate, flocculation efficiency (FE), bacterial community, and EPS profile of axenic and xenic Ettlia cultures were monitored during 46 days of cultivation. For the xenic culture, with a great abundance of growth-promoting and flocculation-inducing bacteria, the biomass density was 18.75% higher and its FE reached 100% in the mid-stationary phase. Moreover, microscopic observation and a quantitative analysis of the EPS revealed the exclusive presence of long filamentous EPS and more compact structure in the xenic Ettlia culture, possibly explaining its better FE. Notwithstanding, for the axenic culture, despite a lower biomass density and reduced abundance of EPS, its FE reached 92.54% in the mid-stationary phase. Thus, the role of the bacterial community was found to be supportive rather than vital for the high settleability of the self-flocculating Ettlia microalgal culture.

The interest of food industry to merchandise natural astaxanthin is growing up. However, it confronts scientific and technological challenges mainly related to its poor water solubility and chemical instability. Here, we present a new quick and efficient green process to simultaneously extract, encapsulate and stabilize astaxanthin from Haematococcus pluvialis. The process is based on the hitherto unexplored combination of supramolecular solvents (SUPRAS), nanostructured liquids generated from amphiphiles through sequential self-assembly and coacervation, and nanostructured lipid carriers (NLCs). These novel nanosystems were characterized by means of dynamic light scattering, AFM and cryoSEM, revealing spherical particles of ∼100 nm. Their antioxidant activity was measured by ORAC (20.6 ± 3.9 μM TE) and α-TEAC (2.92 ± 0.58 µM α-TE) assays and their in vitro capacity to inhibit ROS by DHE probe. Results showed that the SUPRAS-NLCs proposed yield high extraction and encapsulation efficiencies (71 ± 4%) in combination with a remarkable time stability (180 d, 4 °C).

An acidic cultivation strategy was developed to prevent contamination of a lethal fungus Paraphysoderma sedebokerensis in Haematococcus pluvialis culture for astaxanthin production. Instead of generally used neutral pH, an acidic condition (pH 4) was applied to the cultivation, resulting in a significant inhibition of the fungal contamination. This could be ascribed to the acidity-associated denaturation of a surface protein of P. sedebokerensis, which plays an important role in recognition of H. pluvialis. Stress relief strategies including stepwise light irradiation and naturally occurring nitrogen deficiency were employed in the induction stage to minimize the reduction of astaxanthin production caused by acidic pH. Accordingly, an astaxanthin titer of 84.8 mg L−1 was obtained, which is 141-fold of that from the completely contaminated culture and double of that without the stress relief methods. This strategy provides a persistent contamination control method that can be used for practical astaxanthin production by H. pluvialis.

In this study, we estimated the toxicity risks from river sediments that were affected by multiple pollutants in the Haihe River Basin. We used a range of methods to determine the concentrations, bioavailability, and toxicity of a range of metals and contaminants in sediments and sediment porewater and then assessed the ecological risks and toxicity using various multivariate statistical approaches. We found that more than 70% of the samples were toxic. The concentrations of non-ionic ammonia (0.168–9.295 mg L−1) were generally high in the sediment porewater, while the concentrations of bioavailable chromium (Cr) and polycyclic aromatic hydrocarbons (PAHs) were also high in the porewater samples from NW01 and NW02, respectively. We used the toxic unit (TU) approach, based on the bioavailable pollutant concentrations, to determine the toxicity of PAHs, heavy metals, and non-ionic ammonia in river sediments and sediment porewater. We found that non-ionic ammonia was the main source of toxicity for Daphnia magna, and that Cr and zinc were toxic for Pseudokirchneriella subcapitata and Chironomus dilutus. By combining various indexes, we identified the main contributors to the toxicity in sediments collected from rivers affected by multiple pollutants.

The biosorption properties of water-soluble radioactive cesium (137Cs) by microalga Haematococcus pluvialis were evaluated with different cell conditions, and its cesium-uptake rate was compared with that by other microalgae, Chlorella vulgaris and Anabaena sp. Photo-induced H. pluvialis red cyst rapidly removed radioactive cesium from the solution by bioaccumulation. We showed that the effectiveness of 137Cs uptake is dependent on the specific cell condition of even the same microalgal species. While the H. pluvialis red cyst removed almost 95% of the soluble 137Cs in 48 h, both H. pluvialis intermediate cells and C. vulgaris showed 90% uptake efficiency of 137Cs with slow uptake rate. The energy dispersive spectrometer data demonstrated that the cesium uptake acceleration by inducing astaxanthin in H. pluvialis red cyst involves the cesium accumulation through the potassium transport channel. The long-term monitoring experiments of the cesium uptake showed that only 40% of 137Cs remained in collapsed H. pluvialis cell fragments after 12 months.

Astaxanthin and β-carotene are important carotenoids used in numerous pharmaceutical and nutraceutical applications, owing to their vigorous antioxidant properties. The microalgal strains Haematococcus pluvialis and Dunaliella salina accumulate the highest quantities of astaxanthin and β-carotene (up to 7% and 13% dry weight respectively) and are therefore considered as sustainable feedstock for the commercial production of carotenoids. Thus, from an economical perspective, it becomes desirable to optimize recovery of carotenoids from microalgal cells. To this end, here, we have summarized the conventional and modern extraction techniques generally used for the recovery of astaxanthin from Haematococcus pluvialis and β-carotene from Dunaliella salina. Furthermore, we have also discussed the optimum process conditions employed for numerous extraction protocols including solvent extraction, ultrasonic-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE). Overall, our study highlights the sustainability of integrated co-production of biofuels and carotenoids in a biorefinery framework.

The indigenous microalga, Desmodesmus sp. produced alpha-linolenic acid (ALA) rich lipids in response to low temperature and UV treatment. Incubation at 5 °C showed a 1.5 fold increase in lipid content (34% w/w) with 44% ALA fraction of total fatty acids. The UV treatment (UV 60 min) exhibited a 1.4 fold increase in biomass productivity and 1.6 fold increase in lipid content (37% w/w) with ALA fraction as 31% of total fatty acids. The nitrogen stress enhanced the lipid content (39% w/w) with a reduced ALA fraction (18%) of total fatty acids. The UV treated cultures (UV 40 and 60 min) on incubation at 5 °C showed maximum lipid accumulation (59 to 62% w/w) with ALA fraction of total fatty acids as 39 to 42%. The incubation of nutrient-replete and UV treated cultures at low-temperature could therefore be used for the production of ALA-rich lipids in microalgae.

In the green algal order Volvocales (Chlorophyceae), flagellate colonial forms have evolved at least four times. One of these colonial lineages, Tetrabaenaceae-Goniaceae-Volvocaceae (TGV), which belongs to the clade Reinhardtinia, is closely related to several unicellular chlamydomonads in the genera Chlamydomonas and Vitreochlamys. However, the unicellular sister of TGV has not been specified. Here, the largest ever 18S rRNA phylogenetic tree of Reinhardtinia was constructed including several newly isolated chlamydomonads, and a clade (core-Reinhardtinia) including 32 unicellular lineages and three colonial families were recognized. Interrelationships within core-Reinhardtinia were barely resolved in the tree, and therefore combined 18S-atpB-psaA-psaB-psbC-rbcL gene phylogenetic analyses were performed with selected representatives of 29 of the 32 unicellular lineages and three colonial families. The 29 unicellular lineages were clustered into five metaclades and an unassigned lineage; the metaclade that includes Chlamydomonas pila was resolved, with moderate support, as the sister clade to TGV. To examine possible biases from specific gene(s), long-branch taxa, and the heterogeneous base composition, phylogenetic analyses using several smaller data sets were also performed. Light microscopy of C. pila and its relatives indicated that any early steps towards colony evolution appeared after divergence of TGV from the C. pila lineage.

In the present work, quantitative structure-activity relationship (QSAR) models have been developed for ecotoxicity of pharmaceuticals on four different aquatic species namely Pseudokirchneriella subcapitata, Daphnia magna, Oncorhynchus mykiss and Pimephales promelas using genetic algorithm (GA) for feature selection followed by Partial Least Squares regression technique according to the Organization for Economic Co-operation and Development (OECD) guidelines. Double cross-validation methodology was employed for selecting suitable models. Only 2D descriptors were used for capturing chemical information and model building, whereas validation of the models was performed by considering various stringent internal and external validation metrics. Interestingly, models could be developed even without using any LogP terms in contrary to the usual dependence of toxicity on lipophilicity. However, the current manuscript proposes highly robust and more predictive models employing computed logP descriptors. The applicability domain study was performed in order to set a predefined chemical zone of applicability for the obtained QSAR models, and the test compounds falling outside the domain were not taken for further analysis while making a prioritized list. An additional comparison was made with ECOSAR, an online expert system for toxicity prediction of organic pollutants, in order to prove predictability of the obtained models. The obtained robust consensus models were utilized to predict the toxicity of a large dataset of approximately 9300 drug-like molecules in order to prioritize the existing drug-like substances in accordance to their acute predicted aquatic toxicities following a scaling technique. Finally, prioritized lists of 500 most toxic chemicals obtained by respective consensus models and those predicted from ECOSAR tool have been reported.

Plant factories have been developed to replace traditional agriculture, aiming to solve future problems of food availability. However, the nutrient solution in a plant factory is discharged after a single batch of plant cultivation, giving rise to large amounts of waste nutrient solution. Microalgae can be used to treat a wide variety of wastewater and effectively remove excessive nutrients from wastewater. Therefore, the incorporation of microalgal cultivation into a plant factory to treat waste nutrients would be a reasonable approach facilitating removal of waste nutrients with concomitant production of algal biomass. In this study, we isolated novel microalgal species suitable for the growth in waste nutrients from a plant factory and subsequently an underwater LED photobioreactor was constructed being incorporated into the plant factory system. Finally, treated waste nutrient solution was recycled back into the plant factory and successfully used for the cultivation of plant of butterhead lettuce (Lactuca sativa L.).

Silver nanomaterials (AgNMs) of different shapes and sizes are potentially toxic to aquatic organisms. However, studies on the toxicity of AgNMs and on their shape-dependent effects on algae are scarce. The present study evaluated the effects of three AgNMs (silver nanospheres, AgNPs; silver nanowires, AgNWs; silver nanoplates, AgPLs) with different shapes coated with polyvinylpyrrolidone on the growth and photosynthetic performance of an alga, Chlorococcum infusionum . We used growth measurements and determined the photosynthetic parameters based on chlorophyll fluorescence transients in the algal cells exposed to different concentrations of the three AgNMs. The effective concentrations at 50% (EC 50 ) of AgNPs, AgNWs, and AgPLs were calculated to be 0.1, 0.045, and 0.021 mg/L, respectively. The results showed that the toxicity of AgNMs in C. infusionum was in the order, AgPLs (40 nm diameter) > AgNWs (21,000 nm length × 42 nm diameter) > AgNPs (57 nm diameter), based on the decrease in growth and three photosynthetic activities. We propose that the toxic potential of AgNMs is primarily dependent on their diameter and secondarily on their shape. Overall, this study provides, for the first time, a comparison of the growth and photosynthetic activities of C. infusionum exposed to AgNMs of three different shapes.

The two-stage cultivation strategy was optimized in this study to simultaneously promote the growth and lipid accumulation of Tetradesmus obliquus. Results showed that the optimal dual-stress conditions were nitrogen concentration at 25 mg N·L

Three novel hydrophobic deep eutectic solvents (DESs) based on oleic acid and terpenes (thymol, DL-menthol, and geraniol) were prepared, characterized, and used to extract astaxanthin from the microalga Haematococcus pluvialis without any pre-treatment of the cells. The three DES were composed of Generally Recognized As Safe (GRAS) and edible ingredients. All the tested DESs gave astaxanthin recovery values of about 60 and 30% in 6 h if applied on freeze-dried biomass or directly on algae culture, respectively. The carotenoid profile was qualitatively identical to what was obtained by using traditional organic solvents, regardless of the DES used; the monoesters of astaxanthin with C18-fatty acids were the main compounds found in all the carotenoid extracts. The thymol:oleic acid DES (TAO) could preserve astaxanthin content after prolonged oxidative stress (40% of the astaxanthin initially extracted was still present after 13.5 h of light exposure), thanks to the superior antioxidant properties of thymol. The capacity of improving astaxanthin stability combined with the intrinsic safety and edibility of the DES components makes the formulation astaxanthin-TAO appealing for the food ingredients/additives industry.

Induction of copper ion (Cu

Unicellular green biflagellates of the order Volvocales (Chlorophyceae, Chlorophyta) are common inhabitants of various types of habitats, and can also form peculiar interspecific relationships. Most of their morphological diversity has historically been assigned to the two prominent genera Chlamydomonas and Chloromonas. Ongoing reclassification of these algae, aided by molecular phylogenetics, has resulted in numerous newly proposed genera, but there are certainly brand-new taxa awaiting recognition. In this study, based on morphological and ultrastructural observations together with sequence data of the nuclear 18S and ITS2 rDNA and the plastid rbcL gene, we describe Adglutina synurophila gen. et sp. nov., a volvocalean biflagellate isolated from colonies of the golden-brown alga Synura petersenii (Chrysophyceae). Phylogenetic analyses placed Adglutina in the phylogroup Moewusinia as a sister lineage to the acidophilic "Chlamydomonas" species. It is characterised by having oval to broadly ellipsoidal cells with a low keel-shaped papilla and a cup-shaped chloroplast lacking a pyrenoid, but possessing a lateral eyespot of a variable position. The unique set of features, together with its Synura-loving nature, anchor Adglutina as a well distinguishable phylogenetic lineage within the Moewusinia. The novel alga has a widespread distribution; it has been found in three European countries to date.

Seaweeds and their derivatives are important bioresources of natural bioactive compounds. Nutritional studies indicate that dietary fibers derived from seaweeds have great beneficial potentials in human health and can be developed as functional food. Moreover, sulfated polysaccharides are more likely to be the main bioactive components which are widely distributed in various species of seaweeds including Phaeophyceae, Rhodophyceae and Chlorophyceae. The catabolism by gut microbiota of the seaweeds-derived dietary fibers (DFs) may be one of the pivotal pathways of their physiological functions. Therefore, in this review, we summarized the latest results of the physiological characteristics of seaweed-derived dietary fiber and highlighted the roles of sulfated polysaccharides in the potential regulatory mechanisms against disorders. Meanwhile, the effects of different types of seaweed-derived dietary fiber on gut microbiota were discussed. The analysis of the structure–function correlations and gut microbiota related mechanisms and will contribute to further better applications in food and biotherapeutics.

Asterarcys quadricellulare (AQ) is a microalgal species with potential applications in improving the quality of animal feed, and safety studies on this species are lacking. Therefore, this study presents safety data on an industrially cultivated strain of AQ tested using the following Organisation for Economic Co-operation and Development (OECD) guidelines: acute skin irritation in rabbits; skin sensitisation in guinea pigs; acute eye irritation in rabbits; acute oral fixed-dose procedure in rats; and bacterial reverse mutation using the B.N. Ames technique. Results showed that AQ is non-irritant and non-sensitising to skin. AQ caused transient conjunctival lacrimation and redness; however, the scores for these clinical signs translated into low ocular irritation indices and classification of AQ as non-irritant to the eyes. An acute oral dose of AQ (2000 mg/kg) did not cause mortality, change in body weight gain, or any general, functional, and neurobehavioral clinical signs. In five strains of Salmonella typhimurium bacteria, treatment with AQ did not cause biologically or statistically significant changes in the number of revertant colonies, indicating that AQ does not cause mutagenic toxicity. This study demonstrates the safety of a heterotrophically-produced strain of AQ and supports its use as a safe and non-toxic feed ingredient.

Monoraphidium sp. SVMIICT6 was isolated and mixotrophically cultivated in a flat-panel photobioreactor (8 days) to treat synthetic dairy wastewater. COD, nitrates, and phosphates removal efficiencies were 75%, 85%, and 60% respectively. The nutrient removal supported the growth of microalgae in terms of biomass productivity (50 mg L

The antibiotic sulfamethoxazole (SMX) is a pollutant that is widely distributed in the global water environment.This substance has toxic effects on various aquatic organisms. Previous studies on SMX have focused on its acute toxicity towards algae and the changes induced at biological and cellular levels, rather than its biotoxicity and mechanisms at the molecular level. In this study, we investigated the effects of SMX on Scenedesmus obliquus as the model organism by performing transmission electron microscopy and transcriptome sequencing analyses. Exposure to SMX promoted gene expression, resulting in changes to algal cell ultrastructure. The cell walls became blurred, the chloroplast structure was seriously damaged, and the number and volume of mitochondria per cell increased. These changes were related to the inhibition of cell growth, decrease in chlorophyll content, increase in cell membrane permeability, and increased production of reactive oxygen species, which led to increased amounts of the lipid peroxidation product malondialdehyde, and higher activities of antioxidant enzymes. Our results suggest that SMX affects gene expression by influencing non-coding RNA metabolic processes, leading to changes in nuclear structures. Abnormally expressed long non-coding RNAs extensively regulate downstream gene expression through various mechanisms, such as chromatin recombination, thereby promoting tumor occurrence, invasion, and metastasis. This abnormal expression may be an important mechanism underlying the carcinogenic effects of SMX.

Silver-based materials have microbicidal action, photocatalytic activity and electronic properties. The increase in manufacturing and consumption of these compounds, given their wide functionality and application, is a source of contamination to freshwater ecosystems and causes toxicity to aquatic biota. Therefore, for the first time, we evaluated the toxicity of the silver tungstate (α-Ag2WO4), in different morphologies (cube and rod), for the microalga Raphidocelis subcapitata. To investigate the toxicity, we evaluated the growth rate, cell complexity and size, reactive oxygen species (ROS) production and chlorophyll a (Chl a) fluorescence. The α-Ag2WO4 – R (rod) was 1.7 times more toxic than α-Ag2WO4–C (cube), with IC10 and IC50 values of, respectively, 8.68 ± 0.91 μg L−1 and 13.72 ± 1.48 μg L−1 for α-Ag2WO4 – R and 18.60 ± 1.61 μg L−1 and 23.47 ± 1.16 μg L−1 for α-Ag2WO4–C. The release of silver ions was quantified and indicated that the silver ions dissolution from the α-Ag2WO4 – R ranged from 34 to 71%, while the Ag ions from the α-Ag2WO4–C varied from 35 to 97%. The α-Ag2WO4–C induced, after 24 h exposure, the increase of ROS at the lowest concentrations (8.81 and 19.32 μg L−1), whereas the α-Ag2WO4 – R significantly induced ROS production at 96 h at the highest concentration (31.76 μg L−1). Both microcrystal shapes significantly altered the cellular complexity and decreased the Chl a fluorescence at all tested concentrations. We conclude that the different morphologies of α-Ag2WO4 negatively affect the microalga and are important sources of silver ions leading to harmful consequences to the aquatic ecosystem.

The bioaccumulation characteristics and acute toxicity of uranium (U) to Hydrodictyon reticulatum were studied to provide reference for further mechanism and application research. According to an analysis using visual MINTEQ software, the pH change caused by the photosynthesis of H. reticulatum leads to U remaining mainly in the species of UO

A novel integrated extraction technique for high recovery of natural astaxanthin from wet encysted Haematococcus pluvialis (H. pluvialis) is demonstrated. The technique can be used to effectively disrupt the cell wall and perform extraction in a one-pot system without a high-energy, cost intensive pre-drying step. The most suitable green solvent was researched in terms of high extraction yield and astaxanthin recovery. Moreover, an optimized condition for the selected green solvents was determined by varying process parameters, viz., the ball milling speed (100-300 rpm) and time (5-30 min). A high recovery of astaxanthin directly from wet H. pluvialis (30.6 mg/g based on its dry mass) and a high extraction yield (58.2 wt%) were achieved using ethyl acetate at 200 rpm after 30 min. Therefore, compared to its counterparts, the biphasic solvent system plays a key role in achieving high extraction yield and astaxanthin recovery from wet H. pluvialis.

With industrialization, anthropogenic mishandlings have resulted in the discharge of abundant amount of CO2 into the atmosphere. This has triggered an unnatural warming that has dramatically increased the Earth’s temperature in a short duration. This problem can be addressed by the biological conversion of CO2; several studies have been conducted using H. pluvialis culture that produces high value-added materials, such as astaxanthin and omega-3 fatty acids. However, although H. pluvialis has a high market value, the market size is quite small. Because H. pluvialis cells are susceptible to contamination due to its slow growth rate, hence large-scale culture of H. pluvialis without reliable contamination control strategies poses significant risks. This review comprehensively discusses the contamination that occurs during the culturing of H. pluvialis in various culture systems under different culture conditions. The review also discusses the strategies in controlling the biotic contaminants, such as bacteria and fungi.

Chytrids are early diverging lineages of true fungi that reproduce with posteriorly uniflagellate zoospores. In aquatic ecosystems, parasitic chytrids of algae have important ecological roles by influencing the population dynamics of phytoplankton and transferring nutrients and energy from inedible algae to zooplankton via zoospores. Despite their ecological importance, information on parasitic chytrids is lacking in the current systematics of chytrids. Here, we investigated a novel chytrid culture KS100 that parasitizes the green alga, Microglena coccifera (Volvocales). A cross-inoculation experiment revealed that KS100 infection was specific to the genus Microglena. Thallus morphology of KS100 is characterized by spherical or subspherical zoosporangium, which becomes slightly angular during zoospore discharge, 2-3 small and inoperculate pores from where zoospores are discharged, and rhizoids branching at the base that extends in a fan-like shape. This combination of characteristics was distinct from any other known chytrids. In molecular phylogeny, KS100 was placed in the order Rhizophydiales and was distinguished from any known families in the order. Zoospores of KS100 possessed a kinetosome-associated structure whose morphology and positioning were unique among the Rhizophydiales. Based on these results, we describe this chytrid as Collimyces mutans gen. et sp. nov. in the new family Collimycetaceae.

Brassinosteroids play an important role in the plant growth and development as well as in the adaptation of plants to environmental stresses. Studies have shown the effect of 24-epibrassinolide (EBL) in the range of concentrations 0.0001–10 μM on the green unicellular alga Acutodesmus obliquus (Chlorophyceae) during 7 days of cultivation. EBL is an effective stimulator of algal growth as it causes an increase in the number of cells and the contents of selected metabolites such as proteins, monosaccharides, and photosynthetic pigments (chlorophylls, carotenes, and xanthophylls). Furthermore, EBL inhibits the formation of reactive oxygen species such as hydrogen peroxide and oxidative damage as evidenced by a decrease of the lipid peroxidation. The positive effect of EBL resulting from the cellular oxidative state can be alleviated by antioxidants such as ascorbate peroxidase, catalase, superoxide dismutase, and ascorbate.

Oxylipins (OXLs) are bioactive molecules generated by the oxidation of fatty acids that promote the resolution of acute inflammation and prevent chronic inflammatory processes through molecular mechanisms that are not well known. We have previously reported the anti-inflammatory activity of microalgae-derived OXLs and OXL-containing biomass in two inflammatory bowel disease (IBD) models: 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced acute colitis and TNBS-induced recurrent colitis. In this study, we examined the in vitro anti-inflammatory mechanism of action of the most abundant OXLs isolated from Chlamydomonas debaryana (13S-HOTE and 13S-HODE) and Nannochloropsis gaditana (15S-HEPE). These OXLs decreased IL-1β and IL-6 pro-inflammatory cytokines production as well as iNOS and COX-2 expression levels in THP-1 macrophages. In addition, OXLs decreased IL-8 production in HT-29 colon cells, the major chemokine produced by these cells. The interaction of OXLs with NFκB and PPAR-γ signaling pathways was studied by confocal microscopy. In THP-1 macrophages and HT-29 colon cells, stimulated by LPS and TNFα respectively, a pre-treatment with 13S-HOTE, 13S-HODE and 15S-HEPE (100μM) resulted in a lower nuclear presence of NFκB in both cell lines. The study of the subcellular localization of PPAR-γ showed that the treatment of THP-1 and HT-29 cells with these OXLs caused the migration of PPAR-γ into the nucleus. Colocalization analysis of both transcription factors in LPS-stimulated THP-1 macrophages showed that the pre-treatment with 13S-HOTE, 13S-HODE or 15S-HEPE lowered nuclear colocalization similar to control value, and increased cytosolic localization above control level. These results indicate that these OXLs could act as agonist of PPAR-γ and consequently inhibit NFκB signaling pathway activation, thus lowering the production of inflammatory markers, highlighting the therapeutic potential of these OXLs in inflammatory diseases such as IBD.