Susan Løvstad Holdt

Bioactive compounds in seaweed: functional food applications and legislation

Seaweed is more than the wrap that keeps rice together in sushi. Seaweed biomass is already used for a wide range of other products in food, including stabilising agents. Biorefineries with seaweed as feedstock are attracting worldwide interest and include low-volume, high value-added products and vice versa. Scientific research on bioactive compounds in seaweed usually takes place on just a few species and compounds. This paper reviews worldwide research on bioactive compounds, mainly of nine genera or species of seaweed, which are also available in European temperate Atlantic waters, i.e. Laminaria sp., Fucus sp., Ascophyllum nodosum, Chondrus crispus, Porphyra sp., Ulva sp., Sargassum sp., Gracilaria sp. and Palmaria palmata. In addition, Undaria pinnatifida is included in this review as this is globally one of the most commonly produced, investigated and available species. Fewer examples of other species abundant worldwide have also been included. This review will supply fundamental information for biorefineries in Atlantic Europe using seaweed as feedstock. Preliminary selection of one or several candidate seaweed species will be possible based on the summary tables and previous research described in this review. This applies either to the choice of high value-added bioactive products to be exploited in an available species or to the choice of seaweed species when a bioactive compound is desired. Data are presented in tables with species, effect and test organism (if present) with examples of uses to enhance comparisons. In addition, scientific experiments performed on seaweed used as animal feed are presented, and EU, US and Japanese legislation on functional foods is reviewed.

Life cycle assessment of biofuel production from brown seaweed in Nordic conditions

The use of algae for biofuel production is expected to play an important role in securing energy supply in the next decades. A consequential life cycle assessment (LCA) and an energy analysis of seaweed-based biofuel production were carried out in Nordic conditions to document and improve the sustainability of the process. Two scenarios were analyzed for the brown seaweed (Laminaria digitata), namely, biogas production (scenario 1) and bioethanol+biogas production (scenario 2). Potential environmental impact categories under investigation were Global Warming, Acidification and Terrestrial Eutrophication. The production of seaweed was identified to be the most energy intensive step. Scenario 1 showed better performance compared to scenario 2 for all impact categories, partly because of the energy intensive bioethanol separation process and the consequently lower overall efficiency of the system. For improved environmental performance, focus should be on optimization of seaweed production, bioethanol distillation, and management of digestate on land.

Microplate-based method for high-throughput screening of microalgae growth potential

Microalgae cultivation conditions in microplates will differ from large-scale photobioreactors in crucial parameters such as light profile, mixing and gas transfer. Hence volumetric productivity (P(v)) measurements made in microplates cannot be directly scaled up. Here we demonstrate that it is possible to use microplates to measure characteristic exponential growth rates and determine the specific growth rate light intensity dependency (μ-I curve), which is useful as the key input for several models that predict P(v). Nannochloropsis salina and Chlorella sorokiniana specific growth rates were measured by repeated batch culture in microplates supplied with continuous light at different intensities. Exponential growth unlimited by gas transfer or self-shading was observable for a period of several days using fluorescence, which is an order of magnitude more sensitive than optical density. The microplate datasets were comparable to similar datasets obtained in photobioreactors and were used an input for the Huesemann model to accurately predict P(v).

Effective harvesting of the microalgae Chlorella protothecoides via bioflocculation with cationic starch.

In the present work, the flocculation efficiency of cationic starch (Greenfloc 120) was tested on the fresh water microalga Chlorella protothecoides under different conditions (pH and flocculant concentrations). Different concentrations of Greenfloc 120 (0, 2.5, 5, 10, 20, 40 mg L(-1)) were screened against different algal densities (0.44, 0.56 and 0.77 g L(-1)). Once the optimal flocculation concentration had been established (40 mg L(-1) for all different biomasses densities) a more detailed analysis was performed in order to investigate if different pH (4.0, 7.7, and 10.0) could increase the flocculation efficiency of cationic starch. Highest flocculation efficiency without addition of Greenfloc 120 was obtained at pH 10, while in the presence of flocculant, the efficiency increased for all the tested pH values, with a maximum of 98% for pH 7.7 and 10. Cationic starch confirmed to be as an easy to use, efficient and cost-effective flocculant for harvesting of microalgae.

Effect of nitrogen source and acclimatization on specific growth rates of microalgae determined by a high-throughput in vivo microplate autofluorescence method

Specific growth rates (SGR) of freshwater algae species (Chlorella vulgaris, Auxenochlorella protothecoides, and Chlorella sorokiniana) and the marine species Nannochloropsis oculata on various nitrogen sources (ammonium carbonate, ammonium chloride, sodium nitrate, and urea) could be determined by in vivo chlorophyll-a autofluorescence. These preferences could be determined before large pH changes occurred in the media, with no significant difference (P > 0.05) between buffered and non-buffered media. In all algal species, acclimatization was observed with no significant difference (P > 0.05) between SGRs of the second and third cultivations. ANOVA of SGRs in the acclimatized second and third cultivations revealed preferences for nitrogen sources among most of the algae; C. vulgaris preferred sodium nitrate over other nitrogen sources, A. protothecoides adapted to urea after no growth in the first cultivation, and the SGRs of N. oculata showed an aversion for sodium nitrate over other nitrogen sources (P < 0.05).

Algal eating habits of phycologists attending the ISAP Halifax Conference and members of the general public

A short questionnaire was devised during the 4th ISAP Conference in Halifax (2011) to gather some information on the algal eating habits of the participants. Responses were also collected from random members of the general public in Galway and Copenhagen. Most phycologists had eaten algae before (93%), but few conference participants ate it more regularly than per month. Responses of the general public were similar. A probability model tested the likelihood of a participant eating algae. Neither age nor nationality significantly influenced this probability, although gender increased the probability of eating algae regularly by 9% if the participant was male (at the 90% confidence limit). As hypothesised, being a conference attendee highly significantly increased the probability of eating algae by 22%, in comparison with non-conference attendees (i.e. the general public). The type of phycological research studied also had a significant effect. Researchers working with macroalgae were 22% more likely to eat algae, whereas microalgal researchers were 15% less likely to eat algae on a monthly or more regular basis. The main reasons for eating algae by both groups were ‘taste’, followed by ‘other’ (undefined) reasons, whereas the main reason for not eating algae in both groups was a ‘lack of availability’. Phycologists also ate algae for the perceived ‘health benefits’ (36%), whereas few members of the general public chose this option (13%). Consumption of red seaweeds was most popular (60% of phycologists and 71% of the general public), with Porphyra spp. most commonly preferred. In total, 27 genera of macroalgae and microalgae were recorded as eaten by questionnaire participants.

Lipids and Composition of Fatty Acids of Saccharina latissima Cultivated Year-Round in Integrated Multi-Trophic Aquaculture

This study is evaluating the seasonal lipid and fatty acid composition of the brown seaweed Saccharina latissima. Biomass was sampled throughout the year (bi-monthly) at the commercial cultivation site near a fish farm in an integrated multi-trophic aquaculture (IMTA) and at a reference site in Denmark (2013–2014). Generally, there was no difference in the biomass composition between sites; however, significant seasonal changes were found. The lipid concentration varied from 0.62%–0.88% dry weight (DW) in July to 3.33%–3.35% DW in November (p < 0.05) in both sites. The fatty acid composition in January was significantly different from all the other sampling months. The dissimilarities were mainly explained by changes in the relative abundance of 20:5n-3 (13.12%–33.35%), 14:0 (11.07%–29.37%) and 18:1n-9 (10.15%–16.94%). Polyunsaturated fatty acids (PUFA’s) made up more than half of the fatty acids with a maximum in July (52.3%–54.0% fatty acid methyl esters; FAME). This including the most appreciated health beneficial PUFA’s, eicosapentaenoic (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), but also arachidonic (ARA) and stearidonic acid (SDA), which are not found in land vegetables such as cabbage and lettuce. Compared to fat (salmon) and lean fish (cod) this seaweed species contains higher proportions of ARA and SDA, but lower EPA (only cod) and DHA. Conclusively, the season of harvest is important for the choice of lipid quantity and quality, but the marine vegetables provide better sources of EPA, DHA and long-chain (LC)-PUFA’s in general compared to traditional vegetables.

High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

Nannochloropsis salina was grown on a mixture of standard growth media and pre-gasified industrial process water representing effluent from a local biogas plant. The study aimed to investigate the effects of enriched growth media and cultivation time on nutritional composition of Nannochloropsis salina biomass, with a focus on eicosapentaenoic acid (EPA). Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scale using a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large-scale cultivation of Nannochloropsis salina at these conditions could improve the nutritional properties such as EPA, tocopherol, protein and carotenoids compared to laboratory-scale cultivated microalgae. EPA reached 44.2% ± 2.30% of total fatty acids, and α-tocopherol reached 431 ± 28 µg/g of biomass dry weight after 21 days of cultivation. Variations in chemical compositions of Nannochloropsis salina were studied during the course of cultivation. Nannochloropsis salina can be presented as a good candidate for winter time cultivation in Denmark. The resulting biomass is a rich source of EPA and also a good source of protein (amino acids), tocopherols and carotenoids for potential use in aquaculture feed industry.

Variation in growth, yield and protein concentration in Saccharina latissima (Laminariales, Phaeophyceae) cultivated with different wave and current exposures in the Faroe Islands

Ropes seeded directly with Saccharina latissima were deployed at different exposures (sheltered, current exposed and wave exposed) in a Faroese sound and characteristics of growth and quality of the biomass and surroundings were evaluated during the growth season from March to August 2015. Saccharina latissima individuals cultivated at the current exposed location were heavier compared to the individuals cultivated at the other locations; however, the total biomass yield was significantly lower at the current exposed location. The protein and nutritional value of the biomass varied with season but showed no correlation with exposure. The highest protein levels and EAA (essential amino acid) score were measured in the spring (April and May) samples. The amino acid composition was dominated by glutamate followed by aspartic acid; however, this was replaced by methionine in the July samples. Total Kjeldahl nitrogen was significantly higher in May and August compared to July, and the nitrate concentration in the biomass was significantly lower in May and August compared to July. Nitrate was most available in the seawater at the time of deployment (3rd of March) and decreased during spring and summer. Saccharina latissima was cultivated successfully at the sheltered, current exposed and wave exposed locations using a direct seeding method. However, our results indicate that the lower biomass yield at the current exposed cultivation location compared to the sheltered and wave exposed are due to the direct seeding method, and possible limitations using this method need further testing and optimization.