Michael Roleda

Research Scientist

(+47) 405 57 498
michael.roleda@nibio.no

Place
Bodø

Visiting address
Torggården, Kudalsveien 6, NO-8027 Bodø

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Abstract

The potential of seaweeds as alternative protein source was investigated in relation to their amino acid (AA) profiles and the ruminal and total tract digestibility of these AAs. Three red (Mastocarpus stellatus, Palmaria palmata, and Porphyra sp.), four brown (Alaria esculenta, Laminaria digitata, Pelvetia canaliculata, and Saccharina latissima), and two green (Cladophora rupestris. and Ulva sp.) seaweed species were used in this study (hereafter, referred to by Genus name only). All seaweeds were collected in Bodø, Northern Norway, during Spring and Autumn in 2014 and 2015, except Ulva, which was only sampled in Autumn of both years, and Saccharina which was not sampled in Spring 2014. All the samples were studied for AA concentration. Six species (Cladophora, Laminaria, Mastocarpus, Palmaria, Porphyra and Ulva) were selected for the more resource demanding in situ study. Species and season interactively affected the content of total AA in crude protein in different seaweeds investigated (P=0.02), with values ranging from 67.2 for Laminaria in Spring to 90.2 gAA/16 g N for Ulva in Autumn. in situ AA degradability was also species specific. The seasonality of total AA in crude protein of different seaweed species mostly did not affect their ruminal degradability, except for alanine, while species and season interactively affected proline’s ruminal degradability. The total tract degradability showed that for Laminaria and Mastocarpus, methionine followed by leucine, isoleucine, histidine and lysine, were protected against rumen degradation. These protections seemed to be acid labile allowing digestion in the lower digestive tract. However, due to high indigestible fractions, these two seaweeds provided low amounts of AA to the intestines. Total tract AA digestibility values were the highest for Porphyra (906 g/kg) followed by Palmaria (843 g/kg) and the green seaweeds. To conclude, Laminaria and Mastocarpus are beneficial sources for bypass protein supply as they contain AA protected against rumen degradation. Based on their amount of AA and their AA degradability, Porphyra, followed by Palmaria and the green seaweeds (Ulva and Cladophora) can be considered as relevant sources of protein for ruminants.

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Abstract

Marine macrophytes are the foundation of algal forests and seagrass meadows–some of the most productive and diverse coastal marine ecosystems on the planet. These ecosystems provide nursery grounds and food for fish and invertebrates, coastline protection from erosion, carbon sequestration, and nutrient fixation. For marine macrophytes, temperature is generally the most important range limiting factor, and ocean warming is considered the most severe threat among global climate change factors. Ocean warming induced losses of dominant macrophytes along their equatorial range edges, as well as range extensions into polar regions, are predicted and already documented. While adaptive evolution based on genetic change is considered too slow to keep pace with the increasing rate of anthropogenic environmental changes, rapid adaptation may come about through a set of non-genetic mechanisms involving the functional composition of the associated microbiome, as well as epigenetic modification of the genome and its regulatory effect on gene expression and the activity of transposable elements. While research in terrestrial plants demonstrates that the integration of non-genetic mechanisms provide a more holistic picture of a species’ evolutionary potential, research in marine systems is lagging behind. Here, we aim to review the potential of marine macrophytes to acclimatize and adapt to major climate change effects via intraspecific variation at the genetic, epigenetic, and microbiome levels. All three levels create phenotypic variation that may either enhance fitness within individuals (plasticity) or be subject to selection and ultimately, adaptation. We review three of the most important phenotypic variations in a climate change context, including physiological variation, variation in propagation success, and in herbivore resistance. Integrating different levels of plasticity, and adaptability into ecological models will allow to obtain a more holistic understanding of trait variation and a realistic assessment of the future performance and distribution of marine macrophytes. Such multi-disciplinary approach that integrates various levels of intraspecific variation, and their effect on phenotypic and physiological variation, is of crucial importance for the effective management and conservation of seagrasses and macroalgae under climate change.

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Abstract

The effects of convective air-drying at 25, 40, and 70 °C and freeze-drying on the quality of the edible kelp Saccharina latissima to be used for food were investigated. Based on the analysis of the carbohydrate and amino acid profiles, as well as polyphenol, fucoxanthin, and ash contents, no significant differences were detected among sample groups, and air-drying up to 70 °C results in equally nutritious products at shorter processing times. Only the iodine content was found lower in freeze-dried compared to air-dried samples. The swelling capacity of the air-dried samples was significantly lower than in freeze-dried samples, particularly at high temperatures (40 and 70 °C), reflecting alteration of the physico-chemical properties of the seaweed during air-drying (attributed to product shrinkage) and reduced capacity of the final product to rehydrate. Structural differences between air-dried products at 25 and 70 °C may explain the differences in mouthfeel perception (dissolving rate) among the two sample groups observed during a sensory evaluation. Overall, the drying temperature within this range did not alter neither the aroma (i.e. odor) nor the flavor intensity of the product. In food applications where the product’s mechanical properties (e.g. porosity) are essential, freeze-drying, and to a lesser extent, air-drying at low temperatures, will result in higher quality products than air-drying at higher temperatures.

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Abstract

A series of semi-continuous incubation experiments were conducted with the coccolithophore Emiliania huxleyi strain NIWA1108 (Southern Ocean isolate) to examine the effects of five environmental drivers (nitrate and phosphate concentrations, irradiance, temperature, and partial pressure of CO2 (pCO2// on both the physiological rates and elemental composition of the coccolithophore. Here, we report the alteration of the elemental composition of E. huxleyi in response to the changes in these environmental drivers. A series of dose–response curves for the cellular elemental composition of E. huxleyi were fitted for each of the five drivers across an environmentally representative gradient. The importance of each driver in regulating the elemental composition of E. huxleyi was ranked using a semiquantitative approach. The percentage variations in elemental composition arising from the change in each driver between present-day and model-projected conditions for the year 2100 were calculated. Temperature was the most important driver controlling both cellular particulate organic and inorganic carbon content, whereas nutrient concentrations were the most important regulator of cellular particulate nitrogen and phosphorus of E. huxleyi. In contrast, elevated pCO2 had the greatest influence on cellular particulate inorganic carbon to organic carbon ratio, resulting in a decrease in the ratio. Our results indicate that the different environmental drivers play specific roles in regulating the elemental composition of E. huxleyi with wide-reaching implications for coccolithophore-related marine biogeochemical cycles, as a consequence of the regulation of E. huxleyi physiological processes.

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Abstract

This study represents a large-scale investigation into iodine contents in three commercially important and edible seaweed species from the North Atlantic: the brown algae Saccharina latissima and Alaria esculenta, and the red alga Palmaria palmata. Variability among and within species were explored in terms of temporal and spatial variations in addition to biomass source. Mean iodine concentration in bulk seaweed biomass was speciesspecific: Saccharina > Alaria > Palmaria. Iodine contents of Saccharina biomass were similar between years and seasons, but varied significantly between sampling locations and biomass sources. In Alaria and Palmaria, none of the independent variables examined contributed significantly to the small variations observed. Our data suggest that all three species are rich sources of iodine, and only 32, 283, or 2149 mg dry weight of unprocessed dry biomass of Saccharina, Alaria, or Palmaria, respectively, meets the recommended daily intake levels for most healthy humans.

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Abstract

The red seaweed Palmaria palmata has previously been reported to have high protein content high in essential amino acids. To extract the proteins a rigid cell wall consisting mainly of β-(1→4)/β-(1→3)-D-xylans must be disrupted. Different methods have been used to overcome this problem along with various methods used for protein evaluation. In this study, the effect of enzymatic pre-treatment on protein extraction was examined. Both enzymatic hydrolysis with xylanase and protease were tested. The amino acid content of the fractions was examined after extraction. The amino acid composition was similar to what has previously been reported; P. palmata was high in essential amino acids. Accordingly, a nitrogen-to-protein conversion factor was calculated for each fraction individually and protein results were compared with calculation using the proximate 6.25 conversion factor. The nitrogen-to-protein conversion factor varied between fractions but all factors were significantly lower than the popularly used 6.25 indicating that this conversion factor for processed P. palmata is effectively and considerably overestimating the protein content. Enzymatic pre-treatment with xylanase resulted in enhanced amino acid content and successful protein extraction. Enzymatic hydrolysis using protease resulted in higher protein content in the liquid extract compared to hydrolysis with xylanase, due to the release of proteins, peptides, and amino acids. Therefore, hydrolysis with protease is not suitable to extract proteins from P. palmata with the method described within this study but might be an optimal method to examine the bioactivity by extracting the protein hydrolysates. However, the result from this study confirm that hydrolysis with xylanase is a feasible choice to extract proteins of good quality from P. palmata.

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We conducted a series of diagnostic fitness response experiments on the coccolithophore, Emiliania huxleyi, isolated from the Subtropical Convergence east of New Zealand. Dose response curves (i.e., physiological rate vs. environmental driver) were constructed for growth, photosynthetic, and calcification rates of E. huxleyi relative to each of five environmental drivers (nitrate concentration, phosphate concentration, irradiance, temperature, and pCO2). The relative importance of each environmental driver on E. huxleyi rate processes was then ranked using a semi-quantitative approach by comparing the percentage change caused by each environmental driver on the measured physiological metrics under the projected conditions for the year 2100, relative to those for the present day, in the Subtropical Convergence. The results reveal that the projected future decrease in nitrate concentration (33%) played the most important role in controlling the growth, photosynthetic and calcification rates of E. huxleyi, whereas raising pCO2 to 75 Pa (750 ppm) decreased the calcification : photosynthesis ratios to the greatest degree. These findings reveal that other environmental drivers may be equally or more influential than CO2 in regulating the physiological responses of E. huxleyi, and provide new diagnostic information to better understand how this ecologically important species will respond to the projected future changes to multiple environmental drivers.

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Abstract

Seaweeds have potentials as alternative feed for ruminants, but there is a limited knowledge on their nutritive value. Seven seaweed species collected along the coast above the Arctic circle of Norway, both in spring and autumn, were assessed for nutrients and total polyphenols (TEP) content, gas production kinetics and in vitro rumen fermentation in batch cultures of ruminal microorganisms. The seaweeds were three red species (Mastocarpus stellatus, Palmaria palmata and Porphyra sp.), three brown species (Alaria esculenta, Laminaria digitata and Pelvetia canaliculata) and one green species (Acrosiphonia sp.). Additionally, the abundance and diversity of total bacteria, protozoa and archaea in the cultures with the three red seaweeds collected in spring were analyzed by quantitative PCR and PCR-DGGE, respectively. The crude protein (CP) content varied widely. Pelvetia had the greatest (P < 0.001) ether extract (EE) content. Non-structural carbohydrates (NSC) content varied from 135 to 541 g/kg DM with brown seaweeds having the greatest values. Ash and CP contents were higher in spring than in autumn (P = 0.020 and 0.003, respectively), whereas concentrations of EE and NSC were not affected by collecting season (P = 0.208–0.341). The TEP values ranged from 1.46 to 50.3 mg/g dry matter (DM), and differed (P < 0.001) among seaweed species and collecting season, being greater in autumn than in spring. The DM effective degradability (DMED), estimated from gas production parameters for a rumen passage rate of 3.0% per h, ranged from 424 to 652 g/kg, the highest values were recorded for Mastocarpus stellatus and Porphyra sp. The lowest DMED values were registered for Pelvetia canaliculata and Acrosiphonia sp. In 24-h incubations (500 mg DM), Palmaria palmata had the highest (P < 0.05) volatile fatty acids (VFA) and methane production (4.34 and 0.761 mmol, respectively) and the lowest (P < 0.05) final pH values and acetate to propionate ratios (6.57 and 2.34, respectively). There were no differences (P > 0.05) among the other seaweeds in VFA production, but Porphyra sp. had the second highest methane production (P < 0.05; 0.491 mmol) compared with the other seaweeds (0.361 mmol; averaged value). The methane/total VFA ratio was not affected (P > 0.05) by either seaweed species or the collection season. Higher final pH (P < 0.05) and lower (P < 0.05) methane and VFA production, ammonia-N concentrations and DMED values were promoted by the fermentation of seaweed collected in autumn compared with those from spring. Among the red seaweeds, there were no species-specific differences (P > 0.05) in the abundance or the diversity of total bacteria, protozoa and archaea. In the PCR-DGGE analysis, samples were separated by the incubation run for all microbial populations analyzed, but not by seaweed species. The results indicate that seaweed species differ markedly in their in vitro rumen degradation, and that samples collected in autumn had lower rumen degradability than those collected in spring.

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Abstract

Anthropogenic atmospheric emissions of CO2 are responsible for simultaneous ocean warming (OW) and ocean acidification (OA). These global events can have important impacts on marine fleshy macroalgae and coastal ecosystems. To understand the effects of OW and OA on the early life history stages of native (Macrocystis pyrifera) and invasive (Undaria pinnatifida) macroalgae, a multi-factorial experiment was performed to determine the independent and interactive effects of the drivers and the corresponding species-specific responses. Meiospores of M. pyrifera and U. pinnatifida were separately exposed to a 4 × 2 factorial design of seawater pH (pHT 7.20, extreme OA predicted for 2300; pHT 7.65, OA predicted for 2100; pHT 8.03, ambient pH; and pHT 8.40, pre-industrial pH) and temperature (12 °C, seasonal average temperature; and 16 °C, OW predicted for 2100). Over 15 days, different physiological parameters (i.e. meiospore germination, germling growth rate, gametophyte development and sex ratio) were measured. Reduced seawater pH and elevated temperature had independent and significant effects on developmental processes (germling growth rate, and male and female gametophyte sizes were independently greater under OA and OW conditions), but the interaction of the abiotic factors had no effect on any stage of meiospore development of either species. Despite some small differences between species (e.g. sex ratio), results of this experiment suggest that microscopic stages of the native M. pyrifera and the invasive U. pinnatifida will respond similarly to OA and OW.

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Abstract

The absorption of anthropogenic CO 2 by the oceans is causing a reduction in the pH of the surface waters termed ocean acidification (OA). This could have substantial effects on marine coastal environments where fleshy (non-calcareous) macroalgae are dominant primary producers and ecosystem engineers. Few OA studies have focused on the early life stages of large macroalgae such as kelps. This study evaluated the effects of seawater pH on the ontogenic development of meiospores of the native kelp Macrocystis pyrifera and the invasive kelp Undaria pinnatifi da , in south-eastern New Zealand. Meiospores of both kelps were released into four seawater pH treatments (pH T 7.20, extreme OA predicted for 2300; pH T 7.65, OA predicted for 2100; pH T 8.01, ambient pH; and pH T 8.40, pre-industrial pH) and cultured for 15 d. Meiospore germination, germling growth rate, and gametophyte size and sex ratio were monitored and measured. Exposure to reduced pH T (7.20 and 7.65) had pos itive effects on germling growth rate and gametophyte size in both M. pyrifera and U. pinnatifida , whereas, higher pH T (8.01 and 8.40) reduced the gametophyte size in both kelps. Sex ratio of gametophytes of both kelps was biased toward females under all pH T treatm ents, except for U. pinnatifida at pH T 7.65. Germling growth rate under OA was significantly higher in M. pyrifera compared to U. pinnatifida but gametophyte development was equal for both kelps under all seawater pH T treatments, indicating that the microscopic stages of the native M. pyrifera and the invasive U. pinnatifida will respond similarly to OA.

Abstract

The 2015-2018 PROMAC (Energy efficient Processing of Macroalgae in blue-green value chains) is financed by the Norwegian Research Council. The PROMAC consortium is led by Møreforsking AS and consists of both Norwegian (SINTEF, NIBIO, NTNU, NMBU) and European (CEVA, MATIS, SLU)research institutes, as well as industrial partners (TafjordKraftvarme, FelleskjøpetFôrutvikling, Firmenich, LegaseaBiomarine Cluster, The Northern Company, Orkla Foods, Hortimare, Marinox).An advisory panel with public authority and interest groups from the marine, energy and agricultural sectors, also oversee the 4,5Mill EUR project’s relevance in a societal context.The current approaches to meeting the demands for meat and other protein-rich food sources are often associated with damage to natural resources and negative effects on climate, air quality, soils and fresh water availability. Therefore, the PROMAC project (http://promac.no/) investigates an alternative approach for providing food and sources of proteins and energy in animal feed, and health benefits in human food through cultivation of macroalgae. The project focuses on the three macroalgaespecies Alariaesculenta, Saccharinalatissima andPalmariapalmata.The research project (i) assesses variation of raw material composition and quality from both harvested and cultured macroalgae, (ii) develops primary processing methods enhancing desired raw material properties, (iii) establishes fractionation and extraction methods best suited to enrich beneficial proteins or remove undesirable anti-nutrients and (iv) evaluates nutritional and health values of processed macroalgal ingredients for various animal groups and in relation to their distinct digestive systems.PROMAC assesses the costs and benefits of macroalgal products from a value chain perspective (from raw material to primary market) through process-based Life Cycle Assessment (LCA), Material and Energy Flow Analysis (MEFA) and business models. To reduce the substantial energy required for primary processing of macroalgae - organisms characterized by ahigh-water content - PROMAC includes a case study utilizing excess heat from a waste incinerator for primary drying and processing of macroalgae biomass. This case study is integrated into both environmental and economic models.Initialresults identifyingmacroalgae food and feed products (ingredients)and associatedprocessing methods most relevant for commercial applications, will be presented integrated across work packages and subject fields.

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Knowledge of life cycle progression and reproduction of seaweeds transcends pure academic interest. Successful and sustainable seaweed exploitation and domestication will indeed require excellent control of the factors controlling growth and reproduction. The relative dominance of the ploidy-phases and their respective morphologies, however, display tremendous diversity. Consequently, the ecological and endogenous factors controlling life cycles are likely to be equally varied. A vast number of research papers addressing theoretical, ecological and physiological aspects of reproduction have been published over the years. Here, we review the current knowledge on reproductive strategies, trade-offs of reproductive effort in natural populations, and the environmental and endogenous factors controlling reproduction. Given that the majority of ecophysiological studies predate the “-omics” era, we examine the extent to which this knowledge of reproduction has been, or can be, applied to further our knowledge of life cycle control in seaweeds.

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Abstract

The production of accurate and reliable data on copper ecotoxicology of marine algae depends on the use of trace metal clean techniques during experimentation. We reviewed the methodologies used in the literature on copper ecotoxicology of marine macro- and microalgae, specifically the use of trace metal clean procedures such as the labware used (glassware vs. plasticware), methods of cleaning the labware (acid soaking and ultrapure water rinsing), stock solution preparation (copper source and acidification), and measurement and reporting of dissolved copper concentrations. In terms of taxonomic classification, the most studied algal groups were the Phyla Ochrophyta, Bacillariophyta, Rhodophyta, and Chlorophyta. In terms of methodology, ∼50% of the articles did not specify the labware, ∼25% used glassware, and ∼25% plasticware; ∼30% of the studies specified cleaning protocols for labware to remove trace metal impurities; the copper form used to prepare the stock solutions was specified in ∼80% of studies but acidification to stabilise the dissolved copper was performed in only ∼20%; and the dissolved copper concentration was measured in only ∼40% of studies. We discuss the importance of following trace metal clean techniques for the comparison and interpretation of data obtained on copper ecotoxicology in algae.

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Abstract

Copper in low natural concentrations is essential for cell metabolism but in excess it becomes extremely toxic to aquatic life, including to the early life stages of marine macroalgae. This work determined the effects of copper exposure on meiospore development of two kelp species, the native Macrocystis pyrifera and invasive Undaria pinnatifida. After settlement, meiospores were exposed to nominal copper concentrations of control (no added copper), 100, 200, 300 and 400 μg L−1 Cu for 9 days. Inductively coupled plasma mass spectrometry of total dissolved copper (CuT) concentrations in the blanks showed that nominal copper concentrations were reduced to 54, 91, 131 and 171 μg L−1 CuT, respectively, indicating that > 50% of the dissolved copper was adsorbed onto the culture vessel walls. In the media with meiospores, the dissolved copper concentrations decreased to 39, 86, 97 and 148 μg L−1 CuT in M. pyrifera and to 39, 65, 97 and 146 μg L−1 CuT in U. pinnatifida, indicating that 6–15% of the dissolved copper was adsorbed by the cells. For both species, meiospores germinated in all copper treatments, with germination decreasing with increasing copper concentration. However, gametophyte growth and sexual differentiation were arrested under all copper treatments. The effective copper concentration causing 50% of arrested germination (Cu-EC50) was 157 and 231 μg L−1 CuT for M. pyrifera and U. pinnatifida, respectively. The higher Cu-EC50 for U. pinnatifida suggests ecological success for the invasive species in copper-polluted environments; however, the subsequent inhibition of gametogenesis under all copper treatments indicated no difference in copper tolerance between both kelp early life stages. We compare our results with the literature available on the effects of copper on the development of early life stages of brown seaweed (Laminariales and Fucales) and discuss the importance of reporting actual experimental dissolved copper concentrations and the necessity of standardizing the response variables measured in macroalgal copper ecotoxicology.

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Abstract

The use of seaweeds in animal diets is not new. However, little is known about the feed value of seaweed, both in terms of chemical composition and protein digestibility, and regarding variation between species and season. In this study, eight seaweed species of the genus Acrosiphonia, Alaria, Laminaria, Mastocarpus, Palmaria, Pelvetia, Porphyra, and Ulva were sampled in spring (March) and autumn (October and November) 2014 at the coast of Bodø in Northern Norway, and were analysed for chemical composition, in situ rumen degradability and total tract crude protein (CP) digestibility. Ash content in dry matter (DM) was generally high (overall mean 190 g/kg in DM) and varied considerably, between species (P < 0.01) and between seasons (P = 0.02). CP concentration in DM varied both between species (P < 0.0001) and seasons (P < 0.01). Highest CP in DM was found for Porphyra (350 g/kg DM) and lowest for Pelvetia (90 g/kg DM). Spring samples were higher in CP than autumn samples. The effective degradability estimated at 5% rumen passage rate (ED5) of CP varied between species (P < 0.0001) but not between seasons (P = 0.10). The highest ED5 of CP was found for Alaria (550 g/kg CP) and lowest for Ulva (240 g/kg CP). Digestible rumen escape protein (DEP) varied significantly between species (P < 0.0001) but not between seasons (P = 0.06); highest DEP was found for Ulva (530 g/kg CP) and Porphyra (500 g/kg CP). Based on our results, Acrosiphonia, Alaria, Laminaria, Mastocarpus and Palmaria can supply the rumen with high amounts of rumen degradable protein, while Porphyra and Ulva can be used as a source of digestible bypass protein. Pelvetia had a very low degradability and should not be used to feed dairy cows.