Ralf Rautenberger

Research Scientist

(+47) 482 10 194
ralf.rautenberger@nibio.no

Place
Bodø

Visiting address
Torggården, Kudalsveien 6, NO-8027 Bodø

Abstract

During the cultivation of Ulva fenestrata in a land-based aquaculture system, the colonisation of the water tanks’ surfaces and eventually the macroalgal biomass by the biofouling diatom Fragilariopsis oceanica compromises the production process. Since germanium dioxide (GeO2) is an effective growth inhibitor of diatoms, this study aimed to understand how it affects the presence of F. oceanica and the photosynthesis and growth of U. fenestrata as a primary parameter contribution to the biomass production. A toxicological dose-response experiment showed that the diatom’s growth was inhibited at the low GeO2 concentration of 0.014 mg l−1. In contrast, the photosynthetic performances and growth rates of U. fenestrata remained unaffected under a wide GeO2 concentration range (0.022–2.235 mg l−1) in small- and large-scale experiments in 1-l glass beakers and 100-l Plexiglass water tanks, respectively. In the latter, the diatom density in the tanks was reduced by 40 %. The costs arising from the use of GeO2 can range between €2.35 and €8.35  kg−1 fresh weight of produced U. fenestrata biomass under growth conditions resulting in growth rates of 20 and 11.5 % d−1, respectively. GeO2 is an effective agent to control biofouling diatoms such as F. oceanica during the land-based biomass production of U. fenestrata.

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Abstract

Chlorella vulgaris is a freshwater microalga that synthesises large amounts of saturated lipids, which makes it suitable for production of bioenergy and biofuels. Since its cultivation usually requires freshwater, it competes with agriculture, economic development and ecological conservation for this limited natural resource. This study investigated the possibility of the partial replacement of freshwater by seawater (50 %) in the growth medium for a more sustainable biomass and lipid production. Chlorella vulgaris 211-11b was cultivated as shake-flask cultures in Bold's Basal Medium (BBM) formulated with 50 % freshwater and 50 % seawater under photoautotrophic, mixotrophic and heterotrophic conditions for eight days with glucose as organic carbon source in the latter two cases. The alga's best growth performance and highest lipid contents (49 % DW−1), dominated by palmitioleic and oleic acid, occurred under mixotrophic rather than photoautotrophic and heterotrophic conditions. This study demonstrates a more economic and ecologically sustainable biomass and lipid production of C. vulgaris by saving 50 % freshwater, which is available for other purposes.

Abstract

Aquaculture of marine macroalgae is an important part of the world’s food production. In Norway, the fast-growing kelp Saccharina latissima has the highest potential for industrial biomass production. Aquaculture in the country’s fjords is economically more viable for SMEs, supports the development of IMTA and could allow the industry to approach the projected 20 million tons by 2050. However, S. latissima is exposed to a considerable decline in seawater salinity during the growth season, which affects the biomass production. This presentation shows results of industrial R&D projects in which the presence of “low-salinity tolerant” strains of S. latissima in a North Norwegian fjord and their responses to the seasonal salinity decline was studied. In a laboratory-based common garden experiment, sporophytes of S. latissima from different locations in Skjerstadfjorden were cultivated under different salinities for six weeks. Growth and photosynthetic parameters were measured to understand their physiological responses to salinity stress. Then their F1 generation were seeded on ropes and deployed at a commercial aquaculture site in Skjerstadfjorden to study strain-specific differences in biomass production and yield, optimal growth depths and biochemical composition of S. latissima. In addition, the aquaculture site was characterised by measurements of physical and chemical parameters. The projects’ results will help North Norwegian macroalgae producers to improve the biomass production and biochemical composition of S. latissima. These findings could lay the foundation for the development of breeding programmes in Norway and could demonstrate the macroalgae producers in Norway the possibility of establishing aquaculture in fjords.

Abstract

Since the world’s population is increasing, alternative food sources must be tapped. Although algae have a high potential to become a part of our diets due to their favorable nutritional properties, there is a little information on the willingness of consumers in Norway to try algae-made foods. In this paper we used a Norwegian survey to address this question. We constructed an order logistic regression model and predicted conditional probabilities to try algae food. The results show that among the most important aspect for willingness to try food with algae is age, health conscientiousness, and environmental attitudes.

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Abstract

Phototrophic microalgae use light to produce biomass and high-value compounds, such as pigments and polyunsaturated fatty acids (PUFA), for food and feed. These biomolecules can be induced by flashing light during the final growth stage. We tested different exposure times (1–6 days) of flashing light (f = 0.5, 5, 50 Hz; duty cycle = 0.05) on biomass, pigment and fatty acid productivity in Diacronema lutheri and Tetraselmis striata. A three-day exposure to low-frequency (5 Hz) flashing light successfully increased the production of fucoxanthin, diatoxanthin, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids in D. lutheri up to 4.6-fold and of lutein, zeaxanthin and EPA in T. striata up to 1.3-fold compared to that of continuous light. Biomass productivity declined 2-fold for D. lutheri and remained similar for T. striata compared to that of continuous light. Thus, short-term treatments of flashing light may be promising for industrial algal production to increase biomass value.

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Abstract

Seaweed is considered a potentially sustainable source of protein for human consumption, and rapid, accurate methods for determining seaweed protein contents are needed. Seaweeds contain substances which interfere with common protein estimation methods however. The present study compares the Lowry and BCA protein assays and protein determination by N-ratios to more novel spectroscopic methods. Linear regression of the height or the integrated area under the Amide II band of diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) was used to predict seaweed protein with good prediction performance. Partial least squares regression (PLSR) was performed on both DRIFTS and near-infrared (NIR) spectra, with even higher prediction accuracy. Spectroscopy performed similar to or better than the calculated N-ratio of 4.14 for protein prediction. These spectral prediction methods require minimal sample preparation and chemical use, and are easy to perform, making them environmentally sustainable and economically viable for rapid estimation of seaweed protein.

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Abstract

Rapid deterioration of harvested macroalgal biomass is a challenge for macroalgal industry and can be overcome with the inexpensive ensiling preservation. To improve silage quality, Saccharina latissima and Alaria esculenta biomass was subjected to ensiling conditions following a 2 × 4 factorial design, with 2 prewilting treatments (no-prewilting and prewilted to 300 g DM kg−1 fresh biomass) and 4 additive treatments (no additive, formic acid, single and two species of Lactobacillus inoculant), and ensiled for 3 or 12 months at 15 °C. Acetate was the main fermentation product in these seaweed silages. Prewilting reduced the acetate, mannitol, and NH3 content in silages. In S. latissima silages without additives, prewilting led to less acidification (pH = 5.7). Also, prewilting caused protein and phlorotannin degradation. When treated with formic acid, the silage pH was below 4 regardless of the biomass’s moisture content. The use of Lactobacillus spp. inoculants was essential for lactate production in seaweed silages, and it significantly lowered silage pH in S. latissima and prewilted A. esculenta compared to silages with no additives. A high level of the phlorotannin content was preserved (> 90%) in the 3-month A. esculenta silages without prewilting. However, major reduction of antioxidant activity was observed in 12-month silages in both seaweed species. In conclusion, ensiling is a viable method for preserving Alaria and Saccharina biomass. Prewilting restricted silage fermentation, and both formic acid and bacterial additives facilitated silage acidification. However, there was no clear benefit of these treatments in preserving the antioxidant activity.

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Abstract

Marine macrophytes, including seagrasses and macroalgae, form the basis of diverse and productive coastal ecosystems that deliver important ecosystem services. Moreover, western countries increasingly recognize macroalgae, traditionally cultivated in Asia, as targets for a new bio-economy that can be both economically profitable and environmentally sustainable. However, seagrass meadows and macroalgal forests are threatened by a variety of anthropogenic stressors. Most notably, rising temperatures and marine heatwaves are already devastating these ecosystems around the globe, and are likely to compromise profitability and production security of macroalgal farming in the near future. Recent studies show that seagrass and macroalgae can become less susceptible to heat events once they have been primed with heat stress. Priming is a common technique in crop agriculture in which plants acquire a stress memory that enhances performance under a second stress exposure. Molecular mechanisms underlying thermal priming are likely to include epigenetic mechanisms that switch state and permanently trigger stress-preventive genes after the first stress exposure. Priming may have considerable potential for both ecosystem restoration and macroalgae farming to immediately improve performance and stress resistance and, thus, to enhance restoration success and production security under environmental challenges. However, priming methodology cannot be simply transferred from terrestrial crops to marine macrophytes. We present first insights into the formation of stress memories in both seagrasses and macroalgae, and research gaps that need to be filled before priming can be established as new bio-engineering technique in these ecologically and economically important marine primary producers.

Abstract

The commercial cultivation of marine macroalgae is a young and rapidly growing industry sector in Norway. Although it is currently limited to a few brown macroalgae, other species such as the green marine macroalga Ulva fenestrata (formerly Ulva lactuca) has also a high potential for an industrial biomass production, for example to be used for the food marked. However, this process is strongly affected by the presence of marine diatoms transported along with the seawater into the cultivation system of U. fenestrata. These diatoms not only proliferate in the water tanks, they also colonise the green macroalgal biomass with many brown spots, which reduces its value for the food marked significantly. This presentation shows the results of a project that studied the use of germanium dioxide (GeO2) as a known growth inhibitor of diatoms to control their contamination during the biomass production process of U. fenestrata. First, the co-occurring diatom was morphologically identified as Fragilaria sp. using light microscopy. Thereafter, a dose-response experiment was conducted to reveal the concentrations of GeO2, resulting in an effective growth inhibition of Fragilaria sp. Based on this knowledge, the impact of different GeO2 concentrations was studied on how the photophysiolgy (photosynthetic characteristics, pigment patterns) and growth of U. fenestrata are affected in both small-scale (2 L) and large-scale (100 L) cultivation systems. An effective control of the proliferation of Fragilaria sp. during the cultivation process of U. fenestrata may result in the production a high-quality biomass with a high value for the food marked.

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Abstract

As the periodic emission of light pulses by light emitting diodes (LEDs) is known to stimulate growth or induce high value biocompounds in microalgae, this flashing light regime was tested on growth and biochemical composition of the microalgae Nannochloropsis gaditana, Koliella antarctica and Tetraselmis chui. At low flashing light frequencies (e.g., 5 and 50 Hz, Duty cycle = 0.05), a strain-dependent growth inhibition and an accumulation of protein, polyunsaturated fatty acids, chlorophyll or carotenoids (lutein, β-carotene, violaxanthin and neoxanthin) was observed. In addition, a 4-day application of low-frequency flashing light to concentrated cultures increased productivities of eicosapentaenoic acid (EPA) and specific carotenoids up to three-fold compared to continuous or high frequency flashing light (500 Hz, Duty cycle = 0.05). Therefore, applying low-frequency flashing light as finishing step in industrial production can increase protein, polyunsaturated fatty acids or pigment contents in biomass, leading to high-value algal products.

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Abstract

Light attenuation in photobioreactors is a major bottleneck in microalgal production. A possible strategy for artificial light-based microalgal production to deliver light deep inside the culture is through the periodical emission of high intensity light flashes (so-called flashing light). However, our results did not show improved photosynthetic rates compared to continuous light for dilute and concentrated Tetraselmis chui cultures exposed to flashing light with various repetition rates (frequencies 0.01 Hz–1 MHz), light-dark ratios (duty cycles: 0.001–0.7) or time-averaged light intensity (50–1000 μmol s−1 m−2). Likewise, flashing light applied to Chlorella stigmatophora and T. chui batch cultures could not enhance growth. However, we observed flashing light effects at different duty cycles and frequencies, depending on cell acclimation, culture concentration, and light intensity. In conclusion, artificial flashing light does not improve microalgal biomass productivities in photobioreactors, but low frequencies (f < 50 Hz) may be still used to improve light harvesting-associated biomolecules production.

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Abstract

Species of the genus Ulva (Chlorophyta) are regarded as opportunistic organisms, which efficiently adjust their metabolism to the prevailing environmental conditions. In this study changes in chlorophyll‐a fluorescence‐based photoinhibition of photosynthesis, electron transport rates, photosynthetic pigments, lipid peroxidation, total phenolic compounds and antioxidant metabolism were investigated during a diurnal cycle of natural solar radiation in summer (for 12 h) under two treatments: photosynthetically active radiation (PAR: 400‐700 nm) and PAR+ ultraviolet (UV) radiation (280‐700 nm). In presence of PAR alone, Ulva rigida showed dynamic photoinhibition, and photosynthetic parameters and pigment concentrations decreased with the intensification of the radiation. On the other hand, under PAR+UV condition a substantial decline up to 43% was detected and an incomplete fluorescence recovery, also, P‐I curve values remained low in relation to the initial condition. The phenolic compounds increased their concentration only in UV radiation treatments without showing a correlation with the antioxidant activity. SOD and APX activities increased over 2‐fold respect at initial values during the onset of light intensity. In contrast, CAT increased its activity rapidly in response to the radiation stress to reach maxima at 10:00 h and decreasing during solar. The present study suggests that U. rigida is capability to acclimate to natural radiation stress relies on a concerted action of various physiological mechanisms that act at different times of the day and under different levels of environmental stress.

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Abstract

Carbonic anhydrase (CA) plays an important physiological role in all biological systems by accelerating the interconversion of CO2 and HCO3 −. In algae, CA is essential for photosynthesis: external CA (CAext) dehydrates HCO3 −, enhancing the supply of CO2 to the cell surface, and internal CA (CAint) interconverts HCO3 − and CO2 to maintain the inorganic carbon (Ci) pool and supply CO2 to RuBisCO. We frst conducted a literature review comparing the conditions in which CA extraction and measurement have been carried out, using the commonly used Wilbur–Anderson method. We found that the assay has been widely modifed since its introduction in 1948, mostly without being optimized for the species tested. Based on the review, an optimized protocol for measuring CA in Macrocystis pyrifera was developed, which showed that the assay conditions can strongly afect CA activity. Tris–HCl bufer gave the highest levels of CA activity, but phosphate bufer reduced activity signifcantly. Bufers containing polyvinylpyrrolidone (PVP) and dithiothreitol (DTT) stabilized CA. Using the optimized assay, CAext and CAint activities were readily measured in Macrocystis with higher precision compared to the non-optimized method. The CAint activity was 2×higher than CAext, which is attributed to the Ci uptake mechanisms of Macrocystis. This study suggests that the CA assay needs to be optimized for each species prior to experimental work to obtain both accurate and precise results.