Seaweed Hatchery and Cultivation Methods

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Seaweed Hatchery and Cultivation Methods Silje Forbord Zeeland 18-19 September

Outline 1. Development of Saccharina latissima (Phaeophyceae) kelp hatcheries with year-round production of zoospores and juvenile sporophytes on culture ropes for kelp aquaculture 2. Sorus disinfection methods 3. Seedling production and hatchery systems 4. Projects and partners 2

Background A biomass with great potential 3.generation biofuel Food and feed Chemicals Fertilizers and minerals Integrated Multi-Trophic Aquaculture (IMTA) Challenges: Cost efficient Industrial biomass production Areas for cultivation BioRefinery Year-round production

Laminaria life cycle

Induction of sorus Short day (8:16) Low light intensities (100 µmol) Low temperature (10 degrees) Number of weeks? Time of year? January 2010-March 2011

Locations for lab and field studies Trondheim (Norway) Grenaa (Denmark) Sylt (Germany)

Results sorus developement Visible sorus were first evident after 2-3 weeks in Sylt and Grenaa and 6 weeks in Trondheim Around 80% of the sporophytes formed sorus at all three locations

Results sorus developement 6 weeks 11 weeks 9 weeks 12 weeks 10 weeks 14 weeks

Seasonal sorus development Visible sorus at the end of experimental series 70-100% of the treated individual formed sorus High variability during the year Successful at time of year when sorus is naturally lacking in the field

On-growth at sea 2 m 5 m 8 m

Conclusions The majority of the vegetative blades may be reliably converted into sporogenous blades independent of season Some small differences in cultivation procedures between the labs robust method Viable zoospores independent of season Year-round production makes it possible to exploit the plants good growth potential on the most suitable time of the year Not all seasons are favorable for deployment and on-growth in sea Need continous access of zoospores to maintain gametophyte cultures Forbord S., Skjermo J., Arff J., Handå A., Reitan K. I., Bjerregaard R., Lüning K. (2012) Development of Saccharina latissima (Phaeophyceae) kelp hatcheries with year-round production of zoospores and juvenile sporophytes on culture ropes for kelp aquaculture. J Appl Phycol 24:393-399

Sori disinfection in cultivation of Saccharina latissima Aim of the thesis: To find a disinfecting method that relieves sori from diatom contamination, without damaging spores or affect early development of young sporophytes

Chemical survey on diatoms Positive growth when SGR 0.05

Chemical survey on diatoms Positive growth when SGR > 0.05 Acetic acid, Lugol's and Sodium hypochlorite were selected for sori disinfection experiments

Sori disinfection method 1. Disks of mature sori were punched out 2. Sori were bathed in a disinfecting solution for a given time interval 3. Sori were rinse in sterile seawater for 30seconds 4. Sori were rinse in sterile seawater for 30seconds 5. Spores were released

Control treatments Acetic acid Figure 9: Sporophyte density, relative spore density during spore release and presents of diatoms in samples added GeO2 (left) and samples from acetic acid disinfection (right). All samples treated with acetic acid were significant different from the control. Bars are mean ± 1SE, n=6. Line is relative spore density. The above dots are %-replica with diatoms present.

Lugol s solution Sodium hypochlorite * Figure 10: Sporophyte density, relative spore density during spore release and presents of diatoms in samples from sorus disinfected with Lugol s solution (left) sodium hypochlorite (right). Bars are mean ± 1SE, *=Significant different from the control. Line is relative spore density. The above dots are %-replica with diatoms present.

Day 3 Day6 Day8 Day11 GeO 2 (0.1mL L -1 ) Control L-2%-2 S-600ppm-2 Foto: Kaia K. Rød

Day 14 Day16 Day18 Day21 L-2%-2 GeO 2 (0.1mL L -1 ) S-600ppm-2 Control Foto: Kaia K. Rød

Conclusions Sodium hypochlorite-600ppm-2min-10 C and Lugol's solution-2%-2min- 10 C are suitable disinfecting treatments Higher sporophyte density in absent of diatoms Indications of higher sporophyte density when the initial spore density is low Low density gives long sporophytes Foto: Kaia K. Rød

Seedling production

Hatchery system I 6-8 mm thick string Water exchange rate of 0.5 L/min UV treated water 16:8 h light:dark regime Good growth but not very space efficient (27 m rope per tank) Not necessary to attach to motherlines at deployment 22

Hatchery system II 1-2 mm thick string Water exchange rate of 0.5 L/min UV treated water Aeration 16:8 h light:dark regime Space efficient, 720 m string per cylinder Seeded both with spores and gametophytes by spraying

Gametophyte cultures Easy to upscale to an industrial scale Shorten the seedling phase Necessary for breeding work Ongoing experiments with growth medium, hormones, light intensities

Growth medium Medium changed every week: Seawater, F/2 (contaminated), Provasoli Medium changed every second week: Seawater, F/2, Provasoli 25

Projects and partners MacroBiomass Duration: 2010-2012 Partners:, Norwegian University of Science and Technology (NTNU), University of Oslo, Sylter Algenfarm (Germany), Blue Food (Denmark) Financed by the Research Council of Norway SeaBreed Duration: 2011-2013 Partners: SINTEF F&A, Seaweed Energy Solutions, CIIMAR (Portugal), Stolt Seafarm (Spain), NTNU Financed by the Research Council of Norway Exploit Duration: 2012-2014 Partners: SINTEF F&A, Institute of Marine Research, NTNU, Bellona Financed by the Research Council of Norway Norwegian Seaweed Technology Center SINTEF F&A and Materials and Chemistry, NTNU Biology and Biotechnology (main partners) Norwegian and international R&D-institutions and industry (associated) Financed by the Regional Research Council 26

Thank you for your attention! Thanks to: Kaia Kjølbo Rød (SES) Sanna Matheson (NTNU) Klaus Lüning (Sylter Algenfarm) Rasmuss Bjerregård (Blue Food) Jorunn Skjermo, Aleksander Handå, Kristine Braaten Steinhovden, Kjell Inge Reitan and Johanne Arff Norwegian Seaweed Technology Center Welcome to the workshop "Seaweed for biofuels" in Trondheim, 25-26.September 2012! Silje.forbord@sintef.no