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Monosaccharides quantified in porewater samples from coastal vegetated ecosystems

Porewater was taken from 30 to 50 cm depths in saltmarsh, seagrass and unvegetated areas around the German Bight in 2022 and 2023. Up to 9 points per ecosystem were sampled along a transect. Polysaccharides >5kDa were upconcentrated using AMICON-filtration devide and afterwards freeze dried. Dired samples were resuspended in MilliQ-water and acid hydrolysed (1 M HCl, 24 h, 100°C). Monosaccharides were analysed using anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD, ThermoFisher Dionex ICS-5000+ system equipped with a CarboPac PA10 analytical column (2 x 250 mm) and a CarboPac PA10 guard column (2 x 50 mm)), according to Engel et al. (2011).

Historical water chemistry monitoring records from multiple stations between 1959 and 2017

Long-term water-chemistry measurements from multiple Elbe River monitoring stations establish a baseline for carbonate-system variability and were used to assess the alkalinity transport potential. The dataset from 1959 to 1977 was digitized from handwritten notes provided by Dr. Mewius (Kempe 1982). The water chemistry data from 1984 to 2017 (e.g., pH, water temperature, and major ions) was obtained from the Fachinformationssystem (FIS) der FGG Elbe (data source: www.fgg-elbe.de, accessed on 2021-02-26).To generate a single river chemistry time series, data from (Zollenspieker (Strom-km 598,7), Geesthacht (Strom-km 585,9), Schnackenburg (Strom-km 474,5), Boizenburg (Strom-km 559,0), Doemitz (Strom-km 505,0), and Hamburg Waterworks (Strom-km ~623,1) were used. Saturation state of calcite and aragonite were calculated using phreeqpython, a Python wrapper of the PhreeqC engine (Vitens 2021) with pH, water temperature, total alkalinity, and major ions as major input, and phreeqc.dat as database for the thermodynamic data (Parkhurst and Appelo 2013).

Schwerpunktprogramm (SPP) 1689: Climate Engineering: Risiken, Herausforderungen, Möglichkeiten?, Modellvergleichende Analyse von CDR Methoden (CDR-MIA)

Die voranschreitenden, anthropogenen CO2-Emissionen verändern das Klima mit bedrohlichen, weit reichenden und irreversiblen Auswirkungen. Daher steigt das Interesse an sogenannten Carbon Dioxide Removal (CDR) Maßnahmen, um so zusätzlich zur Migration und Adaption, die Möglichkeit negativer Emissionen zu eröffnen. Die potenziellen positiven und negativen Auswirkungen durch CDR sind jedoch nicht ausreichend verstanden und quantifiziert. Das Hauptziel des Projektes ist die Analyse der Experimente aus der 1. Phase des Carbon Dioxide Removal Model Intercomparison Projects (CDR-MIP), um das Potenzial und die Risiken großskaliger CDR Methoden besser bewerten zu können. CDR-MIP ist eine neu gegründete Initiative, die eine Reihe von Erdsystemmodellen zusammenbringt, um CDR in einem einheitlichen Rahmen zu untersuchen. Die erste Projektphase, bestehend aus idealisierten Experimenten zu CO2 Entnahme aus der Atmosphäre, Aufforstung und Ozean-Alkalinisierung. Sie dient der Beantwortung folgender Kernfragen a) Reversibilität der Klimaänderung (z.B. zu heutige oder vorindustrielle CO2 Konzentration in der Atmosphäre) und b) potenzielle Wirksamkeit, Feedbacks, zeitlicher Rahmen und Nebenwirkungen unterschiedlicher CDR Maßnahmen. Die bisherige Arbeit diente der Entwicklung der Struktur des CDR-MIPs und weltweit haben sich einige Modellgruppen dazu bereit erklärt die entsprechenden Simulationen durchzuführen. Das Projekt beruht bislang auf freiwilliger Basis. Das macht eine schnelle Verarbeitung der Ergebnisse unwahrscheinlich. Folglich wird eine gezielte Förderung benötigt, um eine zeitnahe Analyse der Ergebnisse und deren öffentlichen Verbreitung zu gewährleisten. Die Analyseergebnisse sollen darüber hinaus die angenommenen Effektivität von CDR Technologien in den 'Integrated Assessment Model (IAM) - generierten Shared Socioeconomic Pathway (SSP) Szenarien informieren, welche die Forschung und Bewertung des Klimawandels unterstützen. Bislang werden bei in den IAM Simulation mit CDR keine Feedbacks des Kohlenstoffkreislaufes berücksichtigt. Eine Wissenslücke die wir schließen wollen. Wir schlagen vor die Ergebnisse aus CDR-MIP zu nutzen, um eine auf den Feedbacks im Kohlenstoffkreislaufes basierende Discount-Rate zu berechnen, die dann für die Kalibrierung der SSP Szenarien und erneuter Modellläufe in einem IAM genutzt werden kann. Zusätzlich werden neue Experimente erstellt und durchgeführt, um die Reaktion des Klimasystems auf die gleichzeitige Anwendung mehrerer CDR Methoden analysieren zu können. Die Kombination der Methoden basiert auf den gegebenen CDR-MIP Experimenten und beinhaltet z.B. eine Kombination von Aufforstung und der Ozean-Alkalinisierung. Anschließende Analysen ermöglichen den Vergleich der Wirksamkeit und Risiken kombinierter und einzelner CDR Methoden. Die Projektergebnisse würden eine umfassende Bewertung von CDR bieten, die allen Projekten innerhalb des SPP verfügbar gemacht und mit den Projektpartnern iterativ diskutiert werden.

CO2-Entnahme durch Alkalinitätserhöhung - Potenzial, Nutzen und Risiken, Vorhaben: Untersuchung regionaler Implikationen, optimaler Ausbringungskonzepte und Umweltfolgen von Alkalinitätserhöhung sowie Simulationen zur Maßnahmenbewertung für die Nordsee

Specific antibody analysis during 24-days of incubations in mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Incubation set-up during mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Particulate organic carbon measurements during 24-days of incubations in mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Surface active fucoidan concentrations quantified in water samples from different depths and stations in the field

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Dissolved organic carbon measurements during 24-days of incubations in mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Fucoidan concentrations in dissolved and surface active fraction during 24-days of incubations in mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

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