ALBERT

Description: In the paper "Salinity as a tool for strain selection in recirculating land-based production of Ulva spp. from germlings to adults" (Cardoso et al. 2023) there was a need to compare artificial and natural seawater based on their nutrient composition and concentration. This experiment was performed to guarantee that the results from the other experiments in this paper were not caused by the different types of water, considering that before the start of these experiment the original cultures were growing in natural seawater. The artificial seawater (30 PSU) (Seequasal-Salz, Seequasal Salz Production and Trade GmbH, Münster, Germany) used in this experiment was made in the laboratory at the Alfred Wegener Institute (AWI), Bremerhaven, Germany, right before being analysed. The natural seawater (30 PSU ± 2 PSU) was collected by the Research Vessel Uthörn in the surrounding area of Helgoland, Germany. The process of collection of natural seawater was made every week to refill and supplement the tanks at AWI, Bremehaven. Therefore, the time of the collection and specific location are not known. The nutrient concentration analysis was done in October 2022 at AWI. Before the analysis both water types were pasteurized for 4 hours at 99 °C. The samples collected for analysis were filtered (0.2 µm) (Nalgene®, Nalge Nunc International, USA) and separated into falcon tubes (n = 3). The analysis was performed by an auto-analyser (SEAL Analytical, United Kingdom) and the concentrations of Phosphate, Ammonium, Nitrite and Nitrate were measured.

Description: In the paper "Salinity as a tool for strain selection in recirculating land-based production of Ulva spp. from germlings to adults" (Cardoso et al. 2023) the relative growth rates of germlings of different Ulva species and strains were measured to evaluate the impact of different salinity treatments. We hypothesised that, since their early stages, salinity impacts the relative growth rate of the seaweeds and that different strains are adapted to different salinities. With this data it was possible to use salinity as a tool for selecting the strain with the highest and determine the optimal salinity to grow its germlings under a nursery setting to supplement a large-scale production. Four strains were tested (U. lacinulata and U. linza from the NE-Atlantic and U. lacinulata and U. flexuosa from the Mediterranean). The NE- Atlantic strains were collected in the Óbidos Lagoon, Portugal in January 2021 and were cultivated in laboratory conditions in the Alfred Wegener Institute (AWI), Bremerhaven, Germany. The Mediterranean strains belong to an AWI collection and were isolated in 1986 and 1987 (U. flexuosa and U. lacinulata, respectively). The cultivation of the Mediterranean species with the purpose of using them for experiments started in June 2021. To recreate the conditions of a nursery system, the strains were grown previously and during the experiment in a medium of artificial seawater (Seequasal-Salz, Seequasal Salz Production and Trade GmbH, Münster, Germany) enriched with half-stregth Provasoli (PES; Provasoli 1968; modifications: HEPES-buffer instead of TRIS, double concentration of Na₂glycerophosphate; iodine enrichment following Tatewaki, 1966). Three germlings with similar size from each species and population were placed into multi-well plates and subjected to different salinity treatments (10, 15, 20 and 30 PSU) (n = 3). The germlings were cultivated for 3 weeks. Because of the small size of the germlings, measuring the fresh weight was not possible and, in this experiment, pictures of the germling development were taken every week and the area of the germlings was measured with the software Image J (Rasband 2012). The difference in size was later used to determine their relative growth rate.

Description: In the paper "Salinity as a tool for strain selection in recirculating land-based production of Ulva spp. from germlings to adults" (Cardoso et al. 2023) the antioxidant activity (AA) experiment aimed to test a previously selected strain, Ulva lacinulata from the NE-Atlantic, and evaluate the variations in AA under different salinity treatments. We hypothesized that by reducing the salinity level in the recirculating land-based system, it would be possible to optimize and increase the quality of the biomass being produced before it being harvested. The NE-Atlantic seaweed was collected in the Óbidos Lagoon, Portugal in January 2021 and was cultivated in laboratory conditions in the Alfred Wegener Institute (AWI), Bremerhaven, Germany. To recreate the conditions of a large-scale system, the strain was grown previously and during the experiment in a medium of artificial seawater (Seequasal-Salz, Seequasal Salz Production and Trade GmbH, Münster, Germany) enriched with a cheaper commercial fertilizer Blaukorn (COMPO SANA®, Germany). Twelve 1 L beakers were filled with artificial seawater at 4 different salinities (10, 15, 20 and 30 PSU), measured with a refractometer (Atago, Japan) (n = 3). Six discs (2 cm) of U. lacinulata were placed in each beaker and, at different times (0 h, 3 h, 24 h, 120 h, 192 h and 240 h) 1 disc was collected from each beaker to evaluate the antioxidant activity. The AA was determined by the ABTS Radical Cation Decolourisation Assay based on Re et al., 1999, and a Trolox standard curve was created by measuring the absorption of different Trolox concentrations in ethanol after being mixed with ABTS. The absorption of the samples at 734 nm was analysed with a microplate reader (Infinite 200 Microplate Reader, Tecan Trading AG, Männedorf, Switzerland) and the AA results obtained were given in Trolox Equivalent (in µg/mL).

Description: In the paper "Salinity as a tool for strain selection in recirculating land-based production of Ulva spp. from germlings to adults" (Cardoso et al. 2023) it was evaluated the impact of different nutrient sources on the relative growth rates (RELATIVE GROWTH RATE) of four different Ulva strains during their adult stage. Previously to this experiment, two experiments were performed to evaluate the of germlings and adults of the four Ulva strains. Because the aim of these experiments was to reproduce the conditions of a large-scale production and a small-scale nursery, the adult material was supplemented with a cheaper commercial fertilizer Blaukorn (COMPO SANA®, Germany) while the germlings (in the nursery set-up) were supplemented with half-strength Provasoli (PES; Provasoli 1968; modifications: HEPES-buffer instead of TRIS, double concentration of Na₂Glycerophosphate; iodine enrichment following Tatewaki, 1966). Therefore, this experiment was performed to guarantee that the use of different nutrient sources would not impact the results of the former experiments. Four strains were tested (U. lacinulata and U. linza from the NE-Atlantic and U. lacinulata and U. flexuosa from the Mediterranean). The NE- Atlantic strains were collected in the Óbidos Lagoon, Portugal in January 2021 and it was cultivated in laboratory conditions in the Alfred Wegener Institute (AWI), Bremerhaven, Germany from that moment on. The Mediterranean strains belong to an AWI collection and were isolated in 1986 and 1987 (U. flexuosa and U. lacinulata, respectively). The cultivation of the Mediterranean species with the purpose of using them for experiments started in June 2021. Fresh thalli from each species and population were placed into 1 L glass beakers and subjected to one of the two treatments: water enriched with commercial fertilizer Blaukorn or with half-strength Provasoli (n = 3). The experiment ran for 3 weeks. The fresh weight was measured once a week by collecting the seaweed and removing the excess water with absorbent paper three times before weighing the samples (Sartorius, Germany). Every time, each sample was measured 3 times. Based on the fresh weight measured each week, it was possible to calculate the of each strain throughout the experiment.

Description: In the paper "Salinity as a tool for strain selection in recirculating land-based production of Ulva spp. from germlings to adults" (Cardoso et al. 2023) the relative growth rates of adult thalli of different Ulva species and strains were measured to evaluate the impact of different salinity treatments. We hypothesised that salinity impacts the of the seaweeds and that different strains are adapted to different salinities. With this data we aimed to use salinity as a tool for selecting the strain with the highest and determine the optimal salinity to grow it in the future in a recirculating land-based system. Four strains were tested (U. lacinulata and U. linza from the NE-Atlantic and U. lacinulata and U. flexuosa from the Mediterranean). The NE- Atlantic strains were collected in the Óbidos Lagoon, Portugal in January 2021 and were cultivated in laboratory conditions in the Alfred Wegener Institute (AWI), Bremerhaven, Germany. The Mediterranean strains belong to an AWI collection and were isolated in 1986 and 1987 (U. flexuosa and U. lacinulata, respectively). The cultivation of the Mediterranean species with the purpose of using them for experiments started in June 2021. To recreate the conditions of a large-scale system, the strains were grown previously and during the experiment in a medium of artificial seawater (Seequasal-Salz, Seequasal Salz Production and Trade GmbH, Münster, Germany) enriched with a cheaper commercial fertilizer Blaukorn (COMPO SANA®, Germany). A uniform amount of fresh thalli from each species and population were placed into 1 L glass beakers with salinities of 10, 15, 20 and 30 PSU (each condition n = 3) and cultivated over 3 weeks. The fresh weight was measured once a week by collecting the seaweed and removing the excess water with absorbent paper three times before weighing the samples (Sartorius, Germany). Every time, each sample was measured 3 times. Based on the fresh weight measured each week, it was possible to calculate the of each strain throughout the experiment.

Description: The mission of the Mak Pak Scale Up project 2021 present) is to develop a seaweed based food packaging material and to sustainably scale up the production of seaweed, including Ulva spp in a recirculating land based facility supplemented with artificial seawater that requires a selection and optimization of the seaweed of interest The Ulva genus has previously shown high plasticity and capacity to acclimate and develop under broad ranges of environmental conditions, suggesting this may be a promising candidate for production in a RAS with low salinity artificial seawater Selecting an Ulva strain well adapted to such conditions will help reduce the costs of salt and, therefore, optimize production The aim of this study was to determine how salinity can be used and adapted throughout the entire Ulva production to define several key points 1 strain selection 2 nursery and seeding 3 optimized production in adult Ulva and 4 increasing functionality (e g high antioxidant activity) Based on the results of three different experiments, it was possible to conclude how salinity impacts different strains of Ulva (tubular and blade species) from warm temperate environments, during three key moments of their development and with that, conclude which strains would be more suited to be scaled up within the scope of land based production At the same time, the data presented serve as a baseline for further work with Ulva under different cultivation conditions

Description: This work intended to optimise the antioxidant activity (AA) of two strains of Ulva lacinulata (Portuguese and Greek) used for food packaging. The enhancement of AA was attempted through the application of hyposaline stress on Ulva for 10 days in one experiment and stress caused by a high light intensity for 5 days in another. The AA was measured using the ABTS radical decolourisation assay. During the light experiment rapid light curves were conducted with help of a Pulse Amplitude Modulated (PAM) fluorometer (Imaging PAM, Heinz Walz GmbH, Germany) to assess the seaweed’s adaptation to a higher light intensity. The Fv/Fm values, initial slope, maximum electron transport rate and light saturation point of the samples were established and compared with the control group. Hyposaline conditions did not enhance AA of the Greek seaweed significantly. The Greek strain was found to have a significantly higher AA regardless of treatment after five days. An exposure to hyposaline conditions of 10 PSU for ten days resulted in a significant increase in AA of the Portuguese algae. A high light intensity resulted in significantly higher AA after five days of exposure in the Portuguese strain, whereas the seaweed of the Greek strain reached maximum AA after 48 hours regardless of treatment. The analysis of the rapid light curves and Fv/Fm values concluded, that the photosynthetic apparatus was not damaged by the high light intensity.