Natural Benzo/Acetophenones as Leads for New Synthetic Acetophenone Hybrids Containing a 1,2,3-Triazole Ring as Potential Antifouling Agents.

Marine biofouling is a natural process that represents major economic, environmental, and health concerns. Some booster biocides have been used in biofouling control, however, they were found to accumulate in environmental compartments, showing negative effects on marine organisms. Therefore, it is urgent to develop new eco-friendly alternatives. Phenyl ketones, such as benzophenones and acetophenones, have been described as modulators of several biological activities, including antifouling activity (AF). In this work, acetophenones were combined with other chemical substrates through a 1,2,3-triazole ring, a strategy commonly used in Medicinal Chemistry. In our approach, a library of 14 new acetophenone-triazole hybrids was obtained through the copper(I)-catalyzed alkyne-azide cycloaddition "click" reaction. All of the synthesized compounds were evaluated against the settlement of a representative macrofouling species, Mytilus galloprovincialis, as well as on biofilm-forming marine microorganisms, including bacteria and fungi. The growth of the microalgae Navicula sp. was also evaluated after exposure to the most promising compounds. While compounds 6a, 7a, and 9a caused significant inhibition of the settlement of mussel larvae, compounds 3b, 4b, and 7b were able to inhibit Roseobacter litoralis bacterial biofilm growth. Interestingly, acetophenone 7a displayed activity against both mussel larvae and the microalgae Navicula sp., suggesting a complementary action of this compound against macro- and microfouling species. The most potent compounds (6a, 7a, and 9a) also showed to be less toxic to the non-target species Artemia salina than the biocide Econea®. Regarding both AF potency and ecotoxicity activity evaluation, acetophenones 7a and 9a were put forward in this work as promising eco-friendly AF agents.

Simulating the Interplay between the Uptake of Inorganic Phosphate and the Cell Phosphate Metabolism under Phosphorus Feast and Famine Conditions in Chlorella vulgaris.

Using a mathematical simulation approach, we studied the dynamics of the green microalga Chlorella vulgaris phosphate metabolism response to shortage and subsequent replenishing of inorganic phosphate in the medium. A three-pool interaction model was used to describe the phosphate uptake from the medium, its incorporation into the cell organic compounds, its storage in the form of polyphosphates, and culture growth. The model comprises a system of ordinary differential equations. The distribution of phosphorous between cell pools was examined for three different stages of the experiment: growth in phosphate-rich medium, incubation in phosphate-free medium, and phosphate addition to the phosphorus-starving culture. Mathematical modeling offers two possible scenarios for the appearance of the peak of polyphosphates (PolyP). The first scenario explains the accumulation of PolyP by activation of the processes of its synthesis, and the decline in PolyP is due to its redistribution between dividing cells during growth. The second scenario includes a hysteretic mechanism for the regulation of PolyP hydrolysis, depending on the intracellular content of inorganic phosphate. The new model of the dynamics of P pools in the cell allows one to better understand the phenomena taking place during P starvation and re-feeding of the P-starved microalgal cultures with inorganic phosphate such as transient PolyP accumulation. Biotechnological implications of the observed dynamics of the polyphosphate pool of the microalgal cell are considered. An approach enhancing the microalgae-based wastewater treatment method based on these scenarios is proposed.

Pyrolysis of High-Ash Natural Microalgae from Water Blooms: Effects of Acid Pretreatment.

Natural microalgae (NA, cyanobacteria) collected from Taihu Lake (Jiangsu, China) were used for biofuel production through pyrolysis. The microalgae were de-ashed via pretreatment with deionized water and hydrochloric acid, and the samples obtained were noted as 0 M, 0.1 M, 1 M, 2 M, 4 M, 6 M, 8 M, respectively, according to the concentration of hydrochloric acid used in the pretreatment. Pyrolysis experiments were carried out at 500 °C for 2 h. The products were examined by various techniques to identify the influence of the ash on the pyrolysis behavior. The results showed that the ash inhibited the thermal transformation of microalgae. The 2 mol/L hydrochloric acid performed the best in removing ash and the liquid yield increased from 34.4% (NA) to 40.5% (2 M). Metal-oxides (mainly CaO, MgO, Al2O3) in ash promoted the reaction of hexadecanoic acid and NH3 to produce more hexadecanamide, which was further dehydrated to hexadecanenitrile. After acid pretreatment, significant improvement in the selectivity of hexadecanoic acid was observed, ranging from 22.4% (NA) to 58.8% (4 M). The hydrocarbon compounds in the liquid product increased from 12.90% (NA) to 26.67% (2 M). Furthermore, the acid pretreatment enhanced the content of C9-C16 compounds and the HHV values of bio-oil. For natural microalgae, the de-ashing pretreatment before pyrolysis was essential for improving the biocrude yield and quality, as well as the biomass conversion efficiency.

Kinetics Growth and Recovery of Valuable Nutrients from Selangor Peat Swamp and Pristine Forest Soils Using Different Extraction Methods as Potential Microalgae Growth Enhancers.

Soil extracts are useful nutrients to enhance the growth of microalgae. Therefore, the present study attempts for the use of virgin soils from Peninsular Malaysia as growth enhancer. Soils collected from Raja Musa Forest Reserve (RMFR) and Ayer Hitam Forest Reserve (AHFR) were treated using different extraction methods. The total dissolved nitrogen (TDN), total dissolved phosphorus (TDP), and dissolved organic carbon (DOC) concentrations in the autoclave methods were relatively higher than natural extraction with up to 132.0 mg N/L, 10.7 mg P/L, and 2629 mg C/L, respectively for RMFR. The results of TDN, TDP, and DOC suggested that the best extraction methods are autoclaved at 121 °C twice with increasing 87%, 84%, and 95%, respectively. Chlorella vulgaris TRG 4C dominated the growth at 121 °C twice extraction method in the RMRF and AHRF samples, with increasing 54.3% and 14%, respectively. The specific growth rate (µ) of both microalgae were relatively higher, 0.23 d-1 in the Ayer Hitam Soil. This extract served well as a microalgal growth promoter, reducing the cost and the needs for synthetic medium. Mass production of microalgae as aquatic feed will be attempted eventually. The high recovery rate of nutrients has a huge potential to serve as a growth promoter for microalgae.

Omega-3 rich oils from microalgae: A chitosan mediated in situ transesterification method.

Microalgae are a precious source of polyunsaturated fatty acids (PUFA), however extraction is difficult due to the peculiar microalgae cell structure. Here we describe a new method based on the application of chitosan nanoaggregates as CO2 responsive emulsifier, used to promote the swelling of algae cell wall and the formation of a large oil - ethanol interphase area during the ethanolysis. Tests were carried out with Pseudokirchneriella subcapitata and Nannochloropsis sp. at different biomass/ethanol/chitosan ratios. CO2 was added to trigger demulsification to promote an easy recovery of the lipid fraction. The highest yields in PUFA were obtained with Nannochloropsis sp. (207.9 mg/g of oil) using 0.4% wt of chitosan and 1:10 biomass:ethanol ratio; 43.6 mg/g of linolenic acid were obtained from Pseudokirchneriella subcapitata. Overall, because the method employs ethanol, a generally recognized as safe (GRAS) solvent, and food grade additives, it is suited for the preparation of PUFA supplements.

Effect of iron and phosphorus on the microalgae growth in co-culture.

Iron and phosphorus (P) are the important micro- and macro-nutrient for microalgae growth, respectively. However, the effect of iron and P on microalgae growth in co-culture associating with the formation of dominate algae has not been investigated before. In the current study, Anabaene flos-aquae, Chlorella vulgaris and Melosira sp. were co-cultivated under the addition of different initial iron and P to reveal the effect of iron and phosphorus on the growth of microalgae. The results showed that the mean growth rate of A. flos-aquae, C. vulgaris and Melosira was 0.270, 0.261 and 0.062, respectively, indicating that the A. flos-aquae and C. vulgaris algae are liable to be the dominant algae while the growth of Melosira was restrained when co-cultured. The ratio of Fe to P has a significant impact on the growth of microalgae and could be regarded as an indicator of algae growth. Microalgae showed a much more obvious uptake of iron compared to that of P. The information obtained in the current study was useful for the forecast of water quality and the control of microalgae bloom.

Different Classes of Phytohormones Act Synergistically to Enhance the Growth, Lipid and DHA Biosynthetic Capacity of Aurantiochytrium sp. SW1.

In the present study, the impact of eight phytohormones from six different classes on the growth, lipid and docosahexaenoic acid (DHA) biosynthetic capacity of Aurantiochytrium sp. SW1 (SW1) was evaluated. Kinetin (KIN), jasmonic acid (JA) and gibberellic acid (GA) significantly enhanced the growth and DHA production of SW1 by 16%-28% and 66%-84% in comparison to the control, respectively. The synergistic effect of these three phytohormones, evaluated by the response surface methodology (RSM), showed that a combination of 3.6 mg/L GA, 2.0 mg/L KIN and 20.0 mg/L JA further increased the growth and DHA production of SW1 by 16% to 28% and 22% to 36%, respectively, in comparison to the individual supplementation. The synergistic effect of these phytohormones was also shown to be time-dependent, where feeding at 24 h of cultivation led to 15%, 26% and 35% further increments in the biomass, lipid and DHA production in comparison to that of 0 h, respectively. The determination of stress markers, antioxidant enzymes and key enzymes involved in fatty acid biosynthesis aided to elucidate the potential mechanism underlying the improvement of growth and DHA production by SW1 at various times of feeding. Supplementation with the phytohormones at 24 h exhibited the maximum impact on reducing the level of reactive oxygen species (ROS) and malondialdehyde (MDA), as well as augmented the antioxidants (superoxide dismutase and catalase) and key metabolic enzymes involved in lipogenesis (malic, glucose-6-phosphate dehydrogenase and ATP-citrate lyase) in comparison to the control and other time points. This study signifies the potential application of phytohormones for improving the growth, lipid and DHA production in Aurantiochytrium spp.

Microalgae Cultivation Using Screened Liquid Dairy Manure Applying Different Folds of Dilution: Nutrient Reduction Analysis with Emphasis on Phosphorus Removal.

A number of dairies in southern Idaho employed stationary inclined screens to separate large solid particles out of liquid dairy manure. In this way, the total solid content of the liquid dairy manure can drop about 20%. Solids in dairy wastewater cause high turbidities, which could block the incident light, a key factor in the microalgae cultivation process using wastewaters as culture media. In this study, screened liquid dairy manure was used as the microalgae Chlorella vulgaris culture media. The aim was to optimize the dilution folds for the best growth of Chlorella vulgaris and nutrients' reduction with a special focus on phosphorus removal and recovery. Four folds of dilution, designated as 5*, 10*, 15*, 20*, were applied to the liquid dairy manure to alleviate hindrance of the high turbidity together with the high ammonium. Microalgal cultivation removed a significant amount of turbidity and major nutrients. For differently diluted liquid dairy manures, although the initial turbidities varied a lot, the final removal rates were not significantly different, falling in the range of 88.11-91.73%. Chemical oxygen demand (COD) in the 5-fold diluted liquid dairy manure dropped from 6700 to 1200 mg/L, corresponding to a removal rate of 79.81%. For the 10-fold, 15-fold, and 20-fold diluted manures, Chlorella removed around 67-69% of the initial CODs. Total Kjeldahl nitrogen (TKN) was removed at rates ranging from 70.84 to 73.99% from the four differently diluted liquid dairy manures without significant differences. NH4-N was removed most efficiently by 88.92% from the 20-fold diluted liquid dairy manure, and the least at 68.65% from the 5-fold diluted one. Although the original total phosphorus (TP) concentrations were distinctive for each group, the TP removal rates stayed in the range of 52.16 to 65.22%. Scanning electron microscopy (SEM) and energy-dispersive spectrometry (EDS) analysis of the precipitates harvested from the microalgal cultivation suggested possible phosphate precipitate forms. The chelation of Ca or Mg cations by dissolved organic matter (DOM) under alkaline conditions caused by microalgae cultivation could explain the unsatisfactory phosphorus removals observed in this study.

Effect of iron and magnesium addition on population dynamics and high value product of microalgae grown in anaerobic liquid digestate.

In this study, FeSO4 supplementation ranging from 0 to 4.5 mM, and MgSO4 supplementation ranging from 0 to 5.1 mM were investigated to observe the effect on the population dynamics, biochemical composition and fatty acid content of mixed microalgae grown in Anaerobic Liquid Digestate (ALD). Overall, 3.1 mM FeSO4 addition into ALD increased the total protein content 60% and led to highest biomass (1.56 g L-1) and chlorophyll-a amount (18.7 mg L-1) produced. Meanwhile, 0.4 mM MgSO4 addition increased the total carotenoid amount 2.2 folds and slightly increased the biomass amount. According to the microbial community analysis, Diphylleia rotans, Synechocystis PCC-6803 and Chlorella sorokiniana were identified as mostly detected species after confirmation with 4 different markers. The abundance of Chlorella sorokiniana and Synechocystis PCC-6803 increased almost 2 folds both in iron and magnesium addition. On the other hand, the dominancy of Diphylleia rotans was not affected by iron addition while drastically decreased (95%) with magnesium addition. This study helps to understand how the dynamics of symbiotic life changes if macro elements are added to the ALD and reveal that microalgae can adapt to adverse environmental conditions by fostering the diversity with a positive effect on high value product.

Optimization Growth of Spirulina (Arthrospira) Platensis in Photobioreactor Under Varied Nitrogen Concentration for Maximized Biomass, Carotenoids and Lipid Contents.

AIMS AND BACKGROUND: Spirulina (Arthrospira) platensis (SP) microalgae were cultured in Zarrouk Medium (ZM), containing three nitrogen concentrations (N-limited, N-optimal and Nrich medium) in ten liter-photo-bioreactor (10 L PBR) for 15-days, in order to study changes in lipid compounds (total carotenoids and total lipids and their effect on fatty acid profile). Based on US patent, the yield of bioactive compounds (such as gamma-linolenic acid GLA, C18:3) extracted from microalgae biomass, mainly depends on the extraction processes (1). GLA has much attention with respect to its therapeutic properties such as its ability to decrease blood cholesterol levels. METHODS: The impact of the addition of N in cultures of S. platensis in terms of growth, biomasses and induced lipid compounds (total carotenoids and total lipid contents and its fatty acid profile), as well as the Sonication (SON) and Microwave (MIC) process as aiding techniques for lipid extraction compared with a Cold Condition (COL), was examined. GC/MS method was used to determine the fatty acid profile of lipid extract of SP cultures. RESULTS: In all S. platensis tested culture, the SP was growing successfully, with varying degrees. In N-rich media, the highest cell growth rate and biomass yield were obtained compared with that recorded in other cultures. Under an N-limited condition, SP had higher Total Carotenoids (TCAR, 45.54 mg/g dw) and total lipid contents (TL, 29.51%± 1.92 g/100g dw) compared with that recorded either in N-rich (11.2 mg/g dw) or in N-optimal (6.23 mg/g dw) cultures. Thus, SP copes with the N -stress by altering the metabolic pathways towards inducing lipid biosynthesis. To maximize the TL and TCAR extraction yields, from N-limited cultures, a set of operating process was applied including the Sonication (SON) and Microwave (MIC), which were used as aiding techniques for lipid extraction compared with the Cold Condition (COL) techniques. The results showed that the extraction efficiency of the S. platensis TL increased in the following order: MIC (29.51%± 1.92) > SON (25.46% ± 1.65> COL (20.43% ±1.43). In a comparative study for its fatty acid profiles (FAPs) among all SP cultures, lipids were analyzed by GC/MS. The predominant fatty acids (>10%, of total FA) were found to be myristic acid (C14:0, MA), palmitic acid (C16:0, PA) and oleic acid (C18:1). CONCLUSION: The study concluded that the N-limited condition was found to have a strong influence on biomass dry weight and lipid contents and total carotenoids in SP cells compared to either Nrich or N-optimal conditions. The use of sonication and the microwave techniques lead to a great increase in the extraction of lipid contents and in high amount Polyunsaturated Fatty Acids (PUFAs) in N-limited cultures, in particular, the omega-6 (ω 6) and omega-3 (ω 3) of the essential C18 fatty acids. It seems that the SP rich in lipid content with a high amount of GLC produced under nitrogen limitation in PBR conditions can be used as a food additive or as a nutritional supplement.

Perturbation of fatty acid composition, pigments, and growth indices of Chlorella vulgaris in response to silver ions and nanoparticles: A new holistic understanding of hidden ecotoxicological aspect of pollutants.

This study assesses and compares the influence of silver nanoparticles (AgNPs) and silver nitrate (AgNO3) on the fatty acid composition, pigments, and growth indices of Chlorella vulgaris. Toxicity testing was carried at the estimated and/or above predicted environmental concentrations of AgNPs and AgNO3. AgNO3 treatments impaired the population growth of C. vulgaris about 2-183 times more than the respective AgNPs ones. The pigments displayed a concentration-dependent decrease in response to both forms of silver; however, AgNO3 displayed higher severity to the pigments than AgNPs. In exposure to 10 µg L-1 AgNO3, the contents of chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid, respectively, demonstrated a reduction of about 5, 3, 4, and 4 times when compared with the same respective concentration of AgNPs. Total amounts of saturated (∑SFA), monounsaturated (∑MUFA), and polyunsaturated (∑PUFA) fatty acids as well as the ratio of unsaturated to saturated ones (Unsat./Sat.) displayed somewhat similar-concentration responses. ∑SFA exhibited a hormesis response, and ∑MUFA, ∑PUFA, and Unsat./Sat. did a decreasing trend with increasing concentration of AgNPs and AgNO3. Myristoleic acid, nervonic acid, and eicosadienoic acid revealed the highest sensitivity. Pearson analysis illustrated the highest correlation among myristoleic acid, eicosenoic acid, and nervonic acid as well as among palmitic acid, stearic acid, palmitoleic acid, and oleic acid. Taken together, AgNPs and the released ions could disrupt physiological health state of microalgae through perturbation in the fatty acid composition (especially MUFAs and PUFAs) and other macromolecules. These types of bioperturbations could change the good health state of aquatic ecosystems.

Disruption of thin- and thick-wall microalgae using high pressure gases: Effects of gas species, pressure and treatment duration on the extraction of proteins and carotenoids.

Industrial scale microalgal cell disruption requires low cost, high efficiency and structural conservation of biomolecules for biorefinery. Many cultivated microalgae have thick walls and these walls are barriers for efficient cell disruption. Until recently, despite the high biodiversity of microalgae, little attention has been paid to thin-wall microalgal species in the natural environment for the production and recovery of valuable biomolecules. Instead of developing high power cell disruption devices, utilization of thin-wall species would be a better approach. The present paper describes a simple device that was assembled to evaluate the viability and effectiveness of biomolecule extraction from both thin- and thick-wall species as a proof of concept. This device was tested with high-pressure gases including N2, CO2 plus N2, and air as the disruption force. The highest nitrogen pressure, 110 bar, was not able to disrupt the thick-wall microalgal cells. On the other hand, the thin-wall species was disrupted to different degrees using different pressures and treatment durations. In the same treatment duration, higher nitrogen pressure gave better cell disruption efficiency than the lower pressure. However, in the same pressure, longer treatment duration did not give better efficiency than the shorter duration. High pressure CO2 treatments resulted in low soluble protein levels in the media. The best conditions to disrupt the thin-wall microalgal cells were 110 bar N2 or air for 1 min among these tests. In these conditions, not only were the disruption efficiencies high, but also the biomolecules were well preserved.

Ecotoxicological assessment of bauxite residue (red mud) overflow treated by dissolved air flotation (DAF).

In this study, the effluent from the Bayer process of transforming bauxite into alumina, followed by dissolved air flotation (DAF) treatment, was evaluated to identify the best experimental conditions that generate less toxic or nontoxic effluent for discharge. Two freshwater organisms, the Chlorophyceae microalga Raphidocelis subcapitata (96-h algal growth inhibition test), and the microcrustacean Daphnia similis (48-h acute immobility test), were used to analyze the efficiency of effluent treatment by detecting and comparing the toxicity of the treated effluent. Experimental factorial planning used different concentrations of the coagulant ferric chloride and the flocculant Nalco® N-99-005B, and different recycle ratios. The highest recycle ratio (30%) and highest flocculant concentration (150 mg/L) had the best solids removal efficiency, above 90%. Probably the higher concentration of flocculant led to the formation of a more stable froth, promoting an increase in the hydrophobic characteristics of the flocs. Two operating conditions were satisfactory according to acute and chronic bioassays, with removal efficiencies of total suspended solids and turbidity higher than 98%, for both variables. Producers, R. subcapitata microalga, were more sensitive than the primary consumers, D. similis microcrustaceans, indicating the importance of performing tests with different bioindicators.

Phosphorus-Induced Lipid Class Alteration Revealed by Lipidomic and Transcriptomic Profiling in Oleaginous Microalga Nannochloropsis sp. PJ12.

Phytoplankton are primary producers in the marine ecosystem, where phosphorus is often a limiting factor of their growth. Hence, they have evolved strategies to recycle phosphorus by replacing membrane phospholipids with phosphorus-free lipids. However, mechanisms for replacement of lipid classes remain poorly understood. To improve our understanding, we performed the lipidomic and transcriptomic profiling analyses of an oleaginous marine microalga Nannochloropsis sp. PJ12 in response to phosphorus depletion (PD) and replenishing. In this study, by using (liquid chromatography couple with tandem mass spectrometry) LC-MS/MS-based lipidomic analysis, we show that membrane phospholipid levels are significantly reduced upon PD, while phosphorus-free betaine lipid levels are increased. However, levels of phosphorus-free photosynthetic galactolipid and sulfolipid are not increased upon PD, consistent with the reduced photosynthetic activity. RNA-seq-based transcriptomic analysis indicates that enzymes involved in phospholipid recycling and phosphorus-free lipid synthesis are upregulated, supporting the lipidomic analysis. Furthermore, enzymes involved in FASII (type II fatty acid synthesis) elongation cycle upon PD are transcriptionally downregulated. EPA (eicosapentaenoic acid) level decrease upon PD is revealed by both GC-MS (gas chromatography coupled with mass spectrometry) and LC-MS/MS-based lipidomic analyses. PD-induced alteration is reversed after phosphorus replenishing. Taken together, our results suggest that the alteration of lipid classes upon environmental change of phosphorus is a result of remodeling rather than de novo synthesis in Nannochloropsis sp. PJ12.

Natural and non-toxic products from Fabaceae Brazilian plants as a replacement for traditional antifouling biocides: an inhibition potential against initial biofouling.

In this study, we screened for the antifouling activity of 15 species plant extracts from Brazilian the Brazilian Caatinga Fabaceae against the initial colonization of natural marine bacterial biofilm. We also investigated the potential toxicity of extracts against planktonic and benthic non-target organisms. Aqueous extracts of plants collected in the Caatinga biome (PE, Brazil) were prepared and tested at different concentration levels (0, 0.5, 1, 2, 4, and 8 mg mL-1). Natural marine bacterial consortium was inoculated in multi-well plates and incubated with the different treatments for 48 h. The biofilm and planktonic bacterial density and biomass inhibition were evaluated along with biofilm biomass eradication. The extracts that showed the highest bacterial biofilm inhibition were evaluated for toxicity against microalgae and crustaceans. The biofilm and planktonic bacterial inhibition potential were evaluated through flow cytometry and spectrophotometry. The selected treatments were evaluated for their toxicity using the microalgae Chaetoceros calcitrans, the copepod Nitokra sp., and the brine shrimp Artemia salina as bioindicators. Our work demonstrates the biotechnological potential of Fabaceae plant compounds as a safe antifouling alternative. Anadenanthera colubrina var. cebil fruits and Apuleia leiocarpa leaf extracts showed antibiofilm activity (≥ 80%), while Myroxylon peruiferum and Dioclea grandiflora leaf extracts showed antibiotic activity. These extracts were safe to planktonic and benthic non-target organisms. The results of this study point to potential substitutes to highly toxic antifouling paints and shed light on the prospect of a yet to be explored biome for more sustainable alternatives in biofouling research.

Modulation of lipid content and lipid profile by supplementation of iron, zinc, and molybdenum in indigenous microalgae.

The effects of iron (Fe), zinc (Zn), and molybdenum (Mo) on the biomass yield, lipid content, lipid yield, and fatty acid composition of Chlorella sp. NC-MKM, Graesiella emersonii NC-M1, Scenedesmus acutus NC-M2, and Chlorophyta sp. NC-M5 were studied. Among them, G. emersonii NC-M1 recorded the highest percentage increase in lipid content (140.3%) and neutral lipid (50.9%) under Zn-supplemented condition compared to the control. Also, it showed a 105% and 41.88% increase in lipid yield and neutral lipid under Fe-supplemented condition compared to the control. However, Chlorella sp. NC-MKM recorded an elevation in lipid yield (70.3% rise) and neutral lipid (24.32% rise) compared to the control in Mo-supplemented condition. The enhanced production of reactive oxygen species (ROS) and antioxidant enzyme (SOD and POD) under Fe-, Zn-, and Mo-supplemented condition supports the lipid accumulation. FAME analysis showed that the overall percentage of SFA and MUFA increased after the addition of Fe, Zn, and Mo in a culture medium compared to the control which is vital for a good-quality biodiesel. Further, biodiesel properties derived from FAMEs such as CN, SV, IV, CFPP, OS, υ, ρ, and HHV were found in accordance with biodiesel standard.

Metabolomic foundation for differential responses of lipid metabolism to nitrogen and phosphorus deprivation in an arachidonic acid-producing green microalga.

The green oleaginous microalga Lobosphaera incisa accumulates storage lipids triacylglycerols (TAG) enriched in the long-chain polyunsaturated fatty acid arachidonic acid under nitrogen (N) deprivation. In contrast, under phosphorous (P) deprivation, the production of the monounsaturated oleic acid prevails. We compared physiological responses, ultrastructural, and metabolic consequences of L. incisa acclimation to N and P deficiency to provide novel insights into the key determinants of ARA accumulation. Differential responses to nutrient deprivation on growth performance, carbon-to-nitrogen stoichiometry, membrane lipid composition and TAG accumulation were demonstrated. Ultrastructural analyses suggested a dynamic role for vacuoles in sustaining cell homeostasis under conditions of different nutrient availability and their involvement in autophagy in L. incisa. Paralleling ARA-rich TAG accumulation in lipid droplets, N deprivation triggered intensive chloroplast dismantling and promoted catabolic processes. Metabolome analysis revealed depletion of amino acids and pyrimidines, and repression of numerous biosynthetic hubs to favour TAG biosynthesis under N deprivation. Under P deprivation, despite the relatively low growth penalties, the presence of the endogenous P reserves and the characteristic lipid remodelling, metabolic signatures of energy deficiency were revealed. Metabolome adjustments to P deprivation included depletion in ATP and phosphorylated nucleotides, increased levels of TCA-cycle intermediates and osmoprotectants. We conclude that characteristic cellular and metabolome adjustments tailor the adaptive responses of L. incisa to N and P deprivation modulating its LC-PUFA production.

Natural plant extracts as an economical and ecofriendly alternative for harvesting microalgae.

The study investigated the ability of plant based natural coagulants from Azadirachta indica; Ficus indica; Moringa oleifera; Citrus sinensis; Punica granatum and Musa acuminata to harvest the microalgal biomass. Influence of eluent type (water and NaCl) and concentration (1-5 N) on coagulant extraction; coagulant dosage (1-5 g) and volume (20-100 ml); pH (6-12) and algal concentration (0.1-1 g l-1) on harvesting were analyzed. The results obtained were compared with alum and chitosan. FTIR and biochemical analysis confirmed the presence of bioactive compounds to aid coagulation. Biomass removal efficiency of 75.50% was obtained with M. oleifera extracts (8 mg ml-1) at pH 7.5-7.8, within 100 min. The harvesting efficiency increased to 95.76% when 4 mg ml-1M. oleifera extracts was combined with 0.75 mg ml-1 chitosan. The life cycle and cost analysis acknowledged the eco-friendly coagulants as strong alternative for conventional coagulants used in microalgal harvesting, thereby improvising the overall bioprocess.

An anti-algae adsorbent for uranium extraction: l-Arginine functionalized graphene hydrogel loaded with Ag nanoparticles.

Uranium (VI) is very essential element in nuclear technique and the enrichment uranium has attracted lots of attention. In this work, l-Arginine and Ag nanoparticles (AgNPs) functionalized reduced graphene oxide ternary hydrogel composites (Ag-l-Arg-rGH) were successfully synthesized, which combined the insertion of AgNPs with one-step thermal reduction and an assembly of graphene oxide nanosheets, using l-Arginine (l-Arg) as both a functional and cross-linking agents. The Ag-l-Arg-rGH composites exhibited great enhanced sorption capacity. Kinetic data best fitted the pseudo-second-order model. The thermodynamic data fitted the Langmuir isotherm model and the calculated maximum adsorption capacity is 434.78 mg/g. In addition, the anti-algae experimental results indicated adsorbent showed marked algal inhibition with the presence of AgNPs in the Ag-l-Arg-rGH composites. In the simulated seawater experiments, The distribution coefficient (Kd) value of uranium(VI) with other competing ions was 2.41 × 104 mL g-1. Thereby, the Ag-l-Arg-rGH composites possessed a promising potential for the enrichment uranium (VI) from nature seawater.

Al2O3 nanoparticle impact on the toxic effect of Pb on the marine microalga Isochrysis galbana.

The rapid development and application of nanotechnology have led to increasing concern about the environmental implications of released nanomaterials and potential risks to public health and aquatic ecosystems. Information on the joint effect of nanomaterials and co-existing contaminants such as heavy metals is still inadequate. Our work investigated the effect of Al2O3 nanoparticles (NPs; nano-Al2O3) on the toxic effect of Pb in the unicellular marine phytoplankton Isochrysis galbana. Results showed that a dose-response effect of nano-Al2O3 was found. Significant enhancement of fluorescence in cell cytoplasm rather than cell membrane occurred in the presence of nano-Al2O3, indicating that nano-Al2O3 can penetrate cells and affect the fluorescence emitted from the chloropigments inside them. The presence of nano-Al2O3 has no impact on the toxic effect of Pb at an NP concentration of 1 mg/L but increased that at NP concentrations of 10 mg/L and 100 mg/L. A synergistic effect was also found for the toxic effect of Pb in the presence of 10 mg/L nano-Al2O3. The presence of 100 mg/L nano-Al2O3 significantly increased the bio-uptake of Pb in the range of 0.25 mg/L to 2.0 mg/L Pb, and the maximum accumulated Pb in algae can reach up to 18.22 ng/105 cells with 100 mg/L nano-Al2O3 compared with Pb alone at 2.0 mg/L(12.53 ng/105 cells). Inside cells, Pb loaded onto nano-Al2O3 can be more toxic than the same amount of free Pb species. The results of toxicity tests and accumulated Pb in algae imply that, in addition to the total Pb cell content, the bioavailability of Pb inside algae should be taken into consideration in evaluating the joint toxicity effect. Our work enhances understanding of the combined toxicity of NPs and co-existing heavy metals and is of practical significance in the natural environment.