Freeze-dried Nannochloropsis oceanica biomass protects eicosapentaenoic acid (EPA) from metabolization in the rumen of lambs.

Eicosapentaenoic acid (EPA) from freeze-dried biomass of Nannochloropsis oceanica microalgae resists ruminal biohydrogenation in vitro, but in vivo demonstration is needed. Therefore, the present study was designed to test the rumen protective effects of N. oceanica in lambs. Twenty-eight lambs were assigned to one of four diets: Control (C); and C diets supplemented with: 1.2% Nannochloropsis sp. oil (O); 12.3% spray-dried N. oceanica (SD); or 9.2% N. oceanica (FD), to achieve 3 g EPA /kg dry matter. Lambs were slaughtered after 3 weeks and digestive contents and ruminal wall samples were collected. EPA concentration in the rumen of lambs fed FD was about 50% higher than lambs fed SD or O diets. Nevertheless, the high levels of EPA in cecum and faeces of animals fed N. oceanica biomass, independently of the drying method, suggests that EPA was not completely released and absorbed in the small intestine. Furthermore, supplementation with EPA sources also affected the ruminal biohydrogenation of C18 fatty acids, mitigating the shift from the t10 biohydrogenation pathways to the t11 pathways compared to the Control diet. Overall, our results demonstrate that FD N. oceanica biomass is a natural rumen-protected source of EPA to ruminants.

Mineral phosphorus drives glacier algal blooms on the Greenland Ice Sheet.

Melting of the Greenland Ice Sheet is a leading cause of land-ice mass loss and cryosphere-attributed sea level rise. Blooms of pigmented glacier ice algae lower ice albedo and accelerate surface melting in the ice sheet's southwest sector. Although glacier ice algae cause up to 13% of the surface melting in this region, the controls on bloom development remain poorly understood. Here we show a direct link between mineral phosphorus in surface ice and glacier ice algae biomass through the quantification of solid and fluid phase phosphorus reservoirs in surface habitats across the southwest ablation zone of the ice sheet. We demonstrate that nutrients from mineral dust likely drive glacier ice algal growth, and thereby identify mineral dust as a secondary control on ice sheet melting.

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.

Investigation of architecture development and phosphate distribution in Chlorella biofilm by complementary microscopy techniques.

Microalgae biofilms may play an important role in the mitigation and prevention of eutrophication caused by domestic, agricultural and industrial wastewater effluents. Despite their potential, the biofilm development and role in nutrient removal are not well understood. Its clarification requires comprehensive studies of the complex three-dimensional architecture of the biofilm. In this study, we established a multimodal imaging approach to provide key information regarding architecture development and nutrient distribution in the biofilm of two green algae organisms: Chlorella pyrenoidosa and Chlorella vulgaris. Helium ion microscopy (HIM), scanning electron microscopy coupled with energy dispersive X-ray analysis (SEM-EDX) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were employed for i) elucidation of spatial arrangement, ii) elemental mapping and iii) 3D chemical imaging of the biofilm. The fine structure of the algal biofilm was resolved by HIM, the evidence of the accumulation of phosphate in hot spots was provided by SEM-EDX and the localization of phosphate oxides granules throughout the whole sample was clarified by ToF-SIMS. The reported results shed light on the phosphorus distribution during Chlorella's biofilm formation and highlight the potential of such correlative approach to solve fundamental question in algal biotechnology research.

Response of energy microalgae Chlamydomonas reinhardtii to nitrogen and phosphorus stress.

Microalgae can effectively absorb nitrogen (N) and phosphorus (P) in wastewater, while growth characteristics can be affected by such nutrients. The influences of the N and P concentration on growth, biomass yield, protein yield, and cell ultrastructure of Chlamydomonas reinhardtii (C. reinhardtii) were investigated in this study. The results showed that, in the optimum conditions (24-72 mg/L for N and 4.5-13.5 mg/L for P), the final biomass and protein content of C. reinhardtii could reach maximum value, and the cell organelles (chloroplast, mitochondria,etc.) showed good structures with larger chloroplasts, and more and neater thylakoids. However, if the concentration of nutrients was much higher or lower than the optimal value, it would cause adverse effects on the growth of C. reinhardtii, especially in high nitrogen (1000 mg/L) and low phosphorus (0.5 mg/L) conditions. Under these extreme conditions, the ultrastructure of the cells was also damaged significantly as follows: the majority of the organelles were deformed, the chloroplast membrane became shrunken, and the mitochondria became swollen, even partial disintegrated (differing slightly under high-N and low-P conditions); furthermore, it is found that C. reinhardtii was more sensitive to low-P stress. On the basis of these results, our findings have general implications in the application of wastewater treatment.

Boosting TAG Accumulation with Improved Biodiesel Production from Novel Oleaginous Microalgae Scenedesmus sp. IITRIND2 Utilizing Waste Sugarcane Bagasse Aqueous Extract (SBAE).

This investigation utilized sugarcane bagasse aqueous extract (SBAE), a nontoxic, cost-effective medium to boost triacylglycerol (TAG) accumulation in novel fresh water microalgal isolate Scenedesmus sp. IITRIND2. Maximum lipid productivity of 112 ± 5.2 mg/L/day was recorded in microalgae grown in SBAE compared to modified BBM (26 ± 3 %). Carotenoid to chlorophyll ratio was 12.5 ± 2 % higher than in photoautotrophic control, indicating an increase in photosystem II activity, thereby increasing growth rate. Fatty acid methyl ester (FAME) profile revealed presence of C14:0 (2.29 %), C16:0 (15.99 %), C16:2 (4.05 %), C18:0 (3.41 %), C18:1 (41.55 %), C18:2 (12.41), and C20:0 (1.21 %) as the major fatty acids. Cetane number (64.03), cold filter plugging property (-1.05 °C), and oxidative stability (12.03 h) indicated quality biodiesel abiding by ASTM D6751 and EN 14214 fuel standards. Results consolidate the candidature of novel freshwater microalgal isolate Scenedesmus sp. IITRIND2 cultivated in SBAE, aqueous extract made from copious, agricultural waste sugarcane bagasse to increase the lipid productivity, and could further be utilized for cost-effective biodiesel production.

CO2 Fixation, Lipid Production, and Power Generation by a Novel Air-Lift-Type Microbial Carbon Capture Cell System.

An air-lift-type microbial carbon capture cell (ALMCC) was constructed for the first time by using an air-lift-type photobioreactor as the cathode chamber. The performance of ALMCC in fixing high concentration of CO2, producing energy (power and biodiesel), and removing COD together with nutrients was investigated and compared with the traditional microbial carbon capture cell (MCC) and air-lift-type photobioreactor (ALP). The ALMCC system produced a maximum power density of 972.5 mW·m(-3) and removed 86.69% of COD, 70.52% of ammonium nitrogen, and 69.24% of phosphorus, which indicate that ALMCC performed better than MCC in terms of power generation and wastewater treatment efficiency. Besides, ALMCC demonstrated 9.98- and 1.88-fold increases over ALP and MCC in the CO2 fixation rate, respectively. Similarly, the ALMCC significantly presented a higher lipid productivity compared to those control reactors. More importantly, the preliminary analysis of energy balance suggested that the net energy of the ALMCC system was significantly superior to other systems and could theoretically produce enough energy to cover its consumption. In this work, the established ALMCC system simultaneously achieved the high level of CO2 fixation, energy recycle, and municipal wastewater treatment effectively and efficiently.

Nutrient removal and lipid accumulation properties of newly isolated microalgal strains.

In this work, four microalgae including Chlorella sp. SDEC-10, Chlorella ellipsoidea SDEC-11, Scenedesmus bijuga SDEC-12 and Scenedesmus quadricauda SEDC-13 isolated from a local lake have been investigated for the properties of growth, nutrient removal and lipid accumulation in synthetic sewage. Their biomass ranged between 0.4 and 0.5g/L. The total phosphorus removal efficiency of four strains was nearly 100%, but in the case of total nitrogen and ammonium the removal efficiency was relatively low. Their lipid content, ranging from 25.92% to 27.76% and corresponding to the lipid productivity 7.88-18.08mg/L/d, was higher than that obtained in BG-11. Palmitic acid and oleic acid were the predominant compositions found through fatty acids analysis. S. quadricauda SDEC-13 performed best both in nutrient removal and in lipid production among the four strains.

Fast pyrolysis of microalgae remnants in a fluidized bed reactor for bio-oil and biochar production.

In this study, pyrolysis of microalgal remnants was investigated for recovery of energy and nutrients. Chlorella vulgaris biomass was first solvent-extracted for lipid recovery then the remnants were used as the feedstock for fast pyrolysis experiments using a fluidized bed reactor at 500 °C. Yields of bio-oil, biochar, and gas were 53, 31, and 10 wt.%, respectively. Bio-oil from C. vulgaris remnants was a complex mixture of aromatics and straight-chain hydrocarbons, amides, amines, carboxylic acids, phenols, and other compounds with molecular weights ranging from 70 to 1200 Da. Structure and surface topography of the biochar were analyzed. The high inorganic content (potassium, phosphorous, and nitrogen) of the biochar suggests it may be suitable to provide nutrients for crop production. The bio-oil and biochar represented 57% and 36% of the energy content of the microalgae remnant feedstock, respectively.

Nitrogen and phosphorus removal from municipal wastewater effluent using microalgal biofilms.

Microalgal biofilms have so far received little attention as post-treatment for municipal wastewater treatment plants, with the result that the removal capacity of microalgal biofilms in post-treatment systems is unknown. This study investigates the capacity of microalgal biofilms as a post-treatment step for the effluent of municipal wastewater treatment plants. Microalgal biofilms were grown in flow cells with different nutrient loads under continuous lighting of 230 µmol/m(2)/s (PAR photons, 400-700 nm). It was found that the maximum uptake capacity of the microalgal biofilm was reached at loading rates of 1.0 g/m(2)/day nitrogen and 0.13 g/m(2)/day phosphorus. These maximum uptake capacities were the highest loads at which the target effluent values of 2.2 mg/L nitrogen and 0.15 mg/L phosphorus were still achieved. Microalgal biomass analysis revealed an increasing nitrogen and phosphorus content with increasing loading rates until the maximum uptake capacities. The internal nitrogen to phosphorus ratio decreased from 23:1 to 11:1 when increasing the loading rate. This combination of findings demonstrates that microalgal biofilms can be used for removing both nitrogen and phosphorus from municipal wastewater effluent.

Perspective for the use of genetic transformants in order to enhance the synthesis of the desired metabolites: Engineering chloroplasts of microalgae for the production of bioactive compounds.

Eukaryotic microalgae have recently gained particular interest as bioreactors because they provide attractive alternatives to bacterial, yeast, plant and other cell-based systems currently in use. Over the last years there has been considerable progress in genetic engineering technologies for algae. Biotechnology companies start to apply these techniques to alter metabolic pathways and express valuable compounds in different cell compartments. In particular, the eukaryotic unicellular alga Chlamydomonas reinhardtii appears to be a most promising cell factory since high amounts of foreign proteins have been expressed in its chloroplast compartment. For this alga the complete nuclear, plastidal and mitochondrial genome sequences have been determined and databases are available for any searching or cloning requirements. Apart from being easily transformable, stable transgenic strains and production volumes in full containment can be obtained within a relatively short time. Furthermore, C. reinhardtii is a green alga which belongs to the category of organisms generally recognized as safe (GRAS status). Thus, enhancing food with edible algae like Chlamydomonas engineered to (over)produce functional ingredients has the potential to become an important factor in food and feed technologies.