• Energy Research

Two environmental parameters, temperature and light intensity, were independently used as stress modulators to enhance eicosapentaenoic acid (EPA) production by the microalga Nannochloropsis oculata, without hindering biomass production. A sinusoidal approach was used, as environmental conditions were alternated between optimum and stress status in multi-day cycles. Low temperatures (5 and 10 °C) and light intensities (30 and 50 μmol photons/m2/s) were tested. Results revealed that the modulated stress approach used was able to avoid decreases in biomass production. Temperature stress (10 °C) presented the highest impact, increasing EPA content to 12.8 mgEPA/L, 158% more than the amount obtained in optimum (non-modulated) growth conditions at that point in time, while the lower light intensity stress was able to increase to 126% more. It is important to point out that in both cases increases in EPA amounts resulted from increased content in each individual cell and not just from increased biomass contents. KEY POINTS: • Temperature stress (10 °C) presented the highest impact increasing EPA content 158% • Lower light intensity stress was able to increase EPA to 126% more • EPA increased in individual cell contents simultaneous with biomass increase.

Risk assessment of pesticides has been based on direct toxic effects on aquatic organisms. Indirect effects data are taken into account but with limitations, as it is frequently difficult to predict their real impacts in the ecosystems. In this context the main aim of this work was to assess how the exposure to the herbicide pendimethalin (Prowl(®)), under environmentally relevant concentrations, may compromise the nutritional composition of food for a relevant group of primary consumers of freshwater food webs-the daphnids, thus affecting their reproduction performance and subsequently the long-term sustainability of active populations of this grazer. Therefore, Daphnia magna individuals were chronically exposed in a clean medium to a control diet (NCF - i.e., non-contaminated green algae Raphidocelis subcapitata) and to a contaminated diet (CF - i.e., the same monoalgal culture grown in a medium enriched with pendimethalin in a concentration equivalent to the EC20 for growth inhibition of algae), during which reproductive endpoints were assessed. The algae were analysed for protein, carbohydrate and fatty acid content. The chemical composition of R. subcapitata in the CF revealed a slight decrease on total fatty acid levels, with a particular decrease of essential ω9 monounsaturated fatty acids. In contrast, the protein content was high in the CF. D. magna exposed to CF experienced a 16% reduction in reproduction, measured as the total number of offspring produced per female. Additionally, an internal pendimethalin body burden of 4.226μgg(-1) was accumulated by daphnids fed with CF. Hence, although it is difficult to discriminate the contribution of the pesticide (as a toxic agent transferred through the food web) from that of the food with a poor quality-compromised by the same pesticide, there are no doubts that, under environmentally relevant concentrations of pesticides, both pathways may compromise the populations of freshwater grazers in the long term, with consequences in the control of the primary productivity of these systems.

Abstract Nannochloropsis oculata is naturally rich in eicosapentaenoic acid (EPA). To turn this microalga into an economically viable source for commercial applications, extraction efficiency must be achieved. Pursuing this goal, emerging technologies such as high hydrostatic pressure (HHP) and moderate electric fields (MEF) were tested, aiming to increase EPA accessibility and subsequent extraction yields. The innovative approach used in this study combined these technologies and associated tailored, less hazardous different solvent mixtures (SM) with distinct polarity indexes. Although the classical Folch SM with chloroform: methanol (PI 4.4) provided the highest yield concerning total lipids (166.4 mglipid/gbiomass), diethyl ether: ethanol (PI 3.6) presented statistically higher values in terms of EPA per biomass, corresponding to 1.3-fold increase. When SM were used in HHP and MEF, neither technology independently improved EPA extraction yields, although the sequential combination of technologies did result in 62% increment in EPA extraction. Overall, the SM and extraction methodologies tested (HHP—200 MPa, 21 °C, 15 min, followed by MEF processing at 40 °C, 15 min) enabled increased EPA extraction yields from wet N. oculata biomass. These findings are of high relevance for the food and pharmaceutical industries, providing viable alternatives to the “classical” extraction methodologies and solvents, with increased yields and lower environmental impact. Key points • Et2O: EtOH is a less toxic and more efficient alternative to Folch solvent mixture • HHP or MEF per se was not able to significantly increase EPA extraction yield • Combinations of HHP and MEF technologies increased both lipids and EPA yields Graphical abstract

In order to enhance microalgal growth in photobioreactors (PBRs), light requirement is one of the most important parameters to be addressed; light should indeed be provided at the appropriate intensity, duration, and wavelength. Excessive intensity may lead to photo-oxidation and -inhibition, whereas low light levels will become growth-limiting. The constraint of light saturation may be overcome via either of two approaches: increasing photosynthetic efficiency by genetic engineering, aimed at changing the chlorophyll antenna size; or increasing flux tolerance, via tailoring the photonic spectrum, coupled with its intensity and temporal characteristics. These approaches will allow an increased control over the illumination features, leading to maximization of microalgal biomass and metabolite productivity. This minireview briefly introduces the nature of light, and describes its harvesting and transformation by microalgae, as well as its metabolic effects under excessively low or high supply. Optimization of the photosynthetic efficiency is discussed under the two approaches referred to above; the selection of light sources, coupled with recent improvements in light handling by PBRs, are chronologically reviewed and critically compared.